Researcher Profile and Settings

Affiliation

• Research Faculty of Agriculture　Fundamental AgriScience Research　Bioscience and Chemistry

Job Title

• Professor

Degree

• Ph.D.（Hokkaido University）

URL

http://www.agr.hokudai.ac.jp/fbc/sonoyama/

J-Global ID

• 200901023144219900

Research Interests

• 消化管生理学   食品機能化学   Food Biochemistry   Gastrointestinal Physiology and Immunology   

Research Areas

• Life sciences / Food sciences

Educational Organization

•  Bachelor's degree program　School of Agriculture
•  Master's degree program　Graduate School of Agriculture
•  Doctoral (PhD) degree program　Graduate School of Agriculture

Academic & Professional Experience

• 2018/10 - Today Research Faculty of Agriculture, Hokkaido University Laboratory of Food Biochemistry, Division of Fundamental AgriScience Professor
• 2007/04 - 2018/09 - 北海道大学大学院農学研究院応用生命科学部門食品科学分野食品機能化学研究室 准教授
• 2003/06 - 2007/03 北海道大学大学院助教授（農学研究科応用生命科学専攻食品科学講座食品機能化学分野） 助教授
• 2003 - 2007 Associate Professor
• 1999/04 - 2003/03 北海道大学大学院農学研究科助手（応用生命科学専攻食品科学講座食品機能化学分野） 助手
• 1999 - 2003 Research Associate
• 1992/04 - 1999/03 北海道大学農学部助手（生物機能化学科食品機能化学講座) 助手
• 1992 - 1999 Research Associate
• 1988/04 - 1991/01 ライオン株式会社生物科学研究所 研究員
• 1988 - 1991 Researcher

Education

•        - 1992  Hokkaido University
•        - 1992  Hokkaido University  Graduate School, Division of Agriculture
•        - 1988  Hokkaido University
•        - 1988  Hokkaido University  Graduate School, Division of Agriculture
•        - 1986  Hokkaido University  Faculty of Agriculture
•        - 1986  Hokkaido University  Faculty of Agriculture

Association Memberships

• 日本食物繊維学会   日本栄養・食糧学会   日本食品免疫学会   日本農芸化学会   Biotechnology and AgrochemistryThe Japan Society of Nutrition and Food ScienceThe Japanese Society for ImmunologyJapanese Association for Food Immunology   Japan Society for Bioscience   

Research Activities

Published Papers

• Murine fecal microRNAs alter the composition of cultured gut microbiota

  Fumina Ohsaka, Mayuko Yamaguchi, Yuka Teshigahara, Moeka Yasui, Eisuke Kato, Kei Sonoyama

  Biochemical and Biophysical Research Communications 685 149184 - 149184 0006-291X 2023/12
• White adipose tissue undergoes browning during preweaning period in association with microbiota formation in mice.

  Anju Tsukada, Yuko Okamatsu-Ogura, Emi Futagawa, Yuki Habu, Natsumi Takahashi, Mira Kato-Suzuki, Yuko Kato, Satoshi Ishizuka, Kei Sonoyama, Kazuhiro Kimura

  iScience 26 (7) 107239 - 107239 2023/07/21 

  Beige adipocytes are transiently induced during early postnatal period in mice. Previous studies have suggested that, unlike in adults, the induction is independent of the sympathetic nerve activity; however, the mechanism is yet unknown. Here, we showed that beige adipocytes are induced during the preweaning period in association with the formation of microbiota in mice. Alteration of gut microbiota composition in preweaning mice by maternal treatment with antibiotics or high-fat diet feeding substantially suppressed WAT browning. The suppression was also found in pups transplanted cecal microbiota from pups of high-fat diet-fed dams. These treatments reduced the hepatic expression of genes involved in bile acid synthesis and the serum bile acids level. The abundance of Porphyromonadaceae and Ruminococcaceae in microbiota showed a positive and negative correlation with the induction of beige adipocytes, respectively. This finding may provide comprehensive understanding of the association between gut microbiota and adipose tissue development in the neonatal period.
• Cyclic Nigerosylnigerose Attenuates High-Fat Diet-Induced Fat Deposition, Colonic Inflammation, and Abnormal Glucose Metabolism and Modifies Gut Immunoglobulin a Reactivity to Commensal Bacteria.

  Takeshi Tsuruta, Kei Sonoyama, Taisei Miyamoto, Qui D Nguyen, Akiko Mizote, Mao Teraoka, Naoki Nishino

  Molecular nutrition & food research 67 (14) e2200389  2023/07 

  SCOPE: High-fat diet (HFD) intake induces gut dysbiosis, inflammation in the peripheral tissues, and a reduction in immunoglobulin A (IgA) coating of gut bacteria, which is related to HFD-induced insulin resistance (IR). This study evaluates the effect of cyclic nigerosylnigerose (CNN), a dietary fiber that prevents gut inflammation and promotes IgA coating of gut bacteria, on the above-mentioned HFD-induced disorders. METHODS AND RESULTS: Balb/c mice are fed an HFD and administered CNN for 20 weeks. CNN administration reduces mesenteric adipose tissue weight, colonic tumor necrosis factor α (TNFα) mRNA expression, and serum endotoxin levels and ameliorates HFD-induced abnormal glucose metabolism. Additionally, CNN administration promotes gut bacteria-specific IgA secretion and alters IgA reactivity to gut bacteria. The alterations of IgA reactivity to specific bacteria such as Erysipelatoclostridium, Escherichia, Faecalibaculum, Lachnospiraceae genera, and Stenotrophomonas are correlated with mesenteric adipose tissue weight, colonic TNFα mRNA expression, serum endotoxin levels, and a homeostasis model assessment for IR. CONCLUSION: CNN-induced alterations in IgA reactivity to gut bacteria may be related to the suppression of HFD-induced fat deposition, colonic inflammation, endotoxemia, and IR. These observations indicate that dietary fiber that modulates IgA reactivity to gut bacteria may be useful in preventing HFD-induced disorders.
• Consumption of 1-Kestose Upregulates MicroRNA-200 and -192/215 Families in Lamina Propria Leukocytes of the Murine Large Intestine

  Fumina OHSAKA, Daiki HONMA, Yoshihiro KADOTA, Takumi TOCHIO, Kei SONOYAMA

  Journal of Nutritional Science and Vitaminology 69 (2) 150 - 154 0301-4800 2023/04/30
• Role of microRNAs in the crosstalk between the gut microbiota and intestinal immune system

  Kei SONOYAMA, Fumina OHSAKA

  Bioscience of Microbiota, Food and Health 42 (4) 222 - 228 2023
• Administration of <i>Bifidobacterium pseudolongum</i> suppresses the increase of colonic serotonin and alleviates symptoms in dextran sodium sulfate-induced colitis in mice

  Misa TATSUOKA, Riku SHIMADA, Fumina OHSAKA, Kei SONOYAMA

  Bioscience of Microbiota, Food and Health 2023
• Local free fatty acids trigger the expression of lipopolysaccharide-binding protein in murine white adipose tissue

  Manami SEKI, Akiho MIWA, Fumina OHSAKA, Yugo KARATSU, Takeshi TSURUTA, Shingo HINO, Tatsuya MORITA, Kei SONOYAMA

  Bioscience of Microbiota, Food and Health 41 (2) 54 - 65 2022
• Aicda deficiency exacerbates high-fat diet-induced hyperinsulinemia but not gut dysbiosis in mice

  Takeshi Tsuruta, Teresia Aluoch Muhomah, Kei Sonoyama, Qui D. Nguyen, Yurika Takase, Aoi Nishijima, Shiori Himoto, Emiko Katsumata, Naoki Nishino

  Nutrition Research 93 15 - 26 0271-5317 2021/09 [Refereed]
  
• Consumption of indigestible saccharides and administration of Bifidobacterium pseudolongum reduce mucosal serotonin in murine colonic mucosa

  Misa Tatsuoka, Yosuke Osaki, Fumina Ohsaka, Takeshi Tsuruta, Yoshihiro Kadota, Takumi Tochio, Shingo Hino, Tatsuya Morita, Kei Sonoyama

  British Journal of Nutrition 127 (4) 1 - 13 0007-1145 2021/04/14 [Refereed]
   

  SCFA increase serotonin (5-hydroxytryptamine, 5-HT) synthesis and content in the colon in vitro and ex vivo, but little is known in vivo. We tested whether dietary indigestible saccharides, utilised as a substrate to produce SCFA by gut microbiota, would increase colonic 5-HT content in mice. Male C57BL/6J mice were fed a purified diet and water supplemented with 4 % (w/v) 1-kestose (KES) for 2 weeks. Colonic 5-HT content and enterochromaffin (EC) cell numbers were lower in mice supplemented with KES than those without supplementation, while monoamine oxidase A activity and mRNA levels of tryptophan hydroxylase 1 (Tph1), chromogranin A (Chga), Slc6a4 and monoamine oxidase A (Maoa) genes in the colonic mucosa, serum 5-HT concentration and total 5-HT content in the colonic contents did not differ between groups. Caecal acetate concentration and Bifidobacterium pseudolongum population were higher in KES-supplemented mice. Similar trends were observed in mice supplemented with other indigestible saccharides, that is, fructo-oligosaccharides, inulin and raffinose. Intragastric administration of live B. pseudolongum (10⁸ colony-forming units/d) for 2 weeks reduced colonic 5-HT content and EC cell numbers. These results suggest that changes in synthesis, reuptake, catabolism and overflow of 5-HT in the colonic mucosa are not involved in the reduction of colonic 5-HT content by dietary indigestible saccharides in mice. We propose that gut microbes including B. pseudolongum could contribute to the reduction of 5-HT content in the colonic mucosa via diminishing EC cells.
• Gut commensals suppress interleukin-2 production through microRNA-200/BCL11B and microRNA-200/ETS-1 axes in lamina propria leukocytes of murine large intestine

  Fumina Ohsaka, Yugo Karatsu, Yoshihiro Kadota, Takumi Tochio, Naoki Takemura, Kei Sonoyama

  Biochemical and Biophysical Research Communications 534 808 - 814 0006-291X 2021/01 [Refereed]
   

  The role of microRNAs (miRNAs) in how microbiota influence the host intestinal immune system is not fully understood. We compared the expression profiles of miRNAs and mRNAs in lamina propria leukocytes (LPL) in the large intestines of germ-free (GF) and specific pathogen-free (SPF) mice. Microarray analysis revealed different expression profiles of miRNAs and mRNAs between GF and SPF mice. Quantitative real time-PCR (qRT-PCR) showed that the level of miR-200 family members was significantly higher in SPF mice than in GF mice. In silico prediction followed by qRT-PCR suggested that Bcl11b, Ets1, Gbp7, Stat5b, and Zeb1 genes were downregulated by the miR-200 family. Western blotting revealed that the expression of BCL11B and ETS-1, but not ZEB1, in large intestinal LPL was significantly lower in SPF mice than in GF mice. Interleukin (IL)-2 production in cultured LPL upon stimulation with phorbol 12-myristate 13-acetate and ionomycin for 24 h was significantly lower in SPF mice than in GF mice. Conventionalization of GF mice substantially recapitulated SPF mice in terms of the expression of miR-200 family members and their target genes and IL-2 production in large intestinal LPL. Considering that BCL11B and ETS-1 reportedly function as transcription factors to activate the Il2 gene, we propose that the presence of gut commensals suppresses IL-2 production in large intestinal LPL, at least in part through post-transcriptional downregulation of Bcl11b and Ets1 genes by miR-200 family members.
• Expression of serotonin receptor HTR4 in glucagon-like peptide-1-positive enteroendocrine cells of the murine intestine

  Motoshi Okumura, Akihiro Hamada, Fumina Ohsaka, Takeshi Tsuruta, Tohru Hira, Kei Sonoyama

  Pflügers Archiv - European Journal of Physiology 472 (10) 1521 - 1532 0031-6768 2020/10 [Refereed]
   

  Serotonin (5-hydroxytryptamine [5-HT]) synthesized and released in enterochromaffin (EC) cells participates in various functions in the gastrointestinal tract by acting on a diverse range of 5-HT receptors (HTRs) expressed on smooth muscle, enteric neurons, and epithelial cells. We previously observed that genes encoding HTR2A, HTR2B, and HTR4 are expressed in murine intestinal organoids, suggesting the expression of these HTRs in intestinal epithelial cells. The present study investigated the localization of these HTRs in the murine intestine by immunofluorescence staining. HTR2A, HTR2B, and HTR4 localized in individual solitary cells in the epithelium, while HTR2C was observed in the lamina propria. In the epithelium, HTR2A, HTR2B, and HTR4 colocalized with 5-HT, and HTR4 colocalized with glucagon-like peptide 1 (GLP-1) and peptide YY (PYY). Murine intestinal organoids show a colocalization pattern that is similar to in vivo HTR2A and HTR4 with 5-HT, GLP-1, and PYY. Intraperitoneal and intragastric administration of tegaserod, an HTR4 agonist, failed to alter plasma GLP-1 levels in fasted mice. However, intragastric but not intraperitoneal administration of tegaserod reduced dietary lipid-induced increases of plasma GLP-1 levels. This action of tegaserod was inhibited by co-administration of RS39604, an HTR4 antagonist. These results suggest that murine ileal GLP-1/PYY-producing enteroendocrine (EE) cells express HTR4, while 5-HT-producing EC cells express HTR2A, HTR2B, and HTR4. In addition, the observations regarding in vivo GLP-1 secretion suggest that HTR4 signaling in ileal EE cells suppresses dietary lipid-induced GLP-1 secretion. We thus propose that EC and EE cells may interact with each other through paracrine signaling mechanisms.
• Intestinal microbiota transplantation reveals the role of microbiota in dietary regulation of RegIIIβ and RegIIIγ expression in mouse intestine.

  Udomsopagit T, Miwa A, Seki M, Shimbori E, Kadota Y, Tochio T, Sonoyama K

  Biochemical and Biophysical Research Communications 529 (1) 64 - 69 2020/08 [Refereed]
   

  RegIIIβ and RegIIIγ are antimicrobial peptides expressed in intestinal epithelial cells. Expression of these peptides is reportedly decreased by high-fat diet (HFD) and increased by indigestible oligosaccharides in mice. Clearly, these dietary regimens change the structure of intestinal microbiota. We employed an intestinal microbiota transplantation (IMT) to test whether diet-induced changes in the expression of these peptides are mediated by gut microbiota. C57BL/6J mice were fed either a normal-fat diet (NFD), a HFD, or a NFD supplemented with or without 1-kestose (KES), an indigestible oligosaccharide. Ileal RegIIIβ and RegIIIγ mRNA levels were lower in mice receiving IMT from HFD-fed mice than in those receiving NFD-fed mice and higher in mice receiving IMT from KES-supplemented mice than in those receiving the mice without KES supplementation. Western blot analysis showed that serum RegIIIβ levels changed in parallel with the ileal mRNA levels. We propose that HFD- and KES-induced changes in the ileal RegIIIβ and RegIIIγ expression and in the circulating RegIIIβ levels are mediated, at least in part, by intestinal microbiota.
• DSS大腸炎マウスにおけるアネキシン遺伝子A10欠損の影響

  塩原 幸, 田邊 宏基, 園山 慶, 内山 孝司, 山本 達朗

  日本栄養・食糧学会大会講演要旨集 (公社)日本栄養・食糧学会 74回 203 - 203 2020/04
• Application of Recombinase-Based In Vivo Expression Technology to Bifidobacterium longum subsp. longum for Identification of Genes Induced in the Gastrointestinal Tract of Mice.

  Hiroka Koguchi, Natsumi Ishigami, Mikiyasu Sakanaka, Kako Yoshida, Sayaka Hiratou, Mina Shimada, Satoru Fukiya, Kei Sonoyama, Atsushi Yokota

  Microorganisms 8 (3) 2020/03/13 [Refereed][Not invited]
   

  Bifidobacteria are one of the major components in human gut microbiota and well-known as beneficial microbes. However, clarification of commensal mechanisms of bifidobacteria in the intestines is still ongoing, especially in the presence of the gut microbiota. Here, we applied recombinase-based in vivo expression technology (R-IVET) using the bacteriophage P1 Cre/loxP system to Bifidobacterium longum subsp. longum 105-A (B. longum 105-A) to identify genes that are specifically expressed in the gastrointestinal tract of conventionally raised mice. Oral administration of the genomic DNA library of B. longum 105-A to conventionally raised mice resulted in the identification of 73 in vivo-induced genes. Four out of seven tested genes were verified in vivo-specific induction at least in the cecum by quantitative reverse transcription PCR. Although there is still room for improvement of the system, our findings can contribute to expanding our understanding of the commensal behavior of B. longum in the gut ecosystem.
• Murine intestinal organoids resemble intestinal epithelium in their microRNA profiles

  Fumina Ohsaka, Kei Sonoyama

  Bioscience, Biotechnology, and Biochemistry 82 (9) 1560 - 1567 0916-8451 2018/09/02 [Refereed]
   

  Intestinal organoids were established as an ex vivo model of the intestinal epithelium. We investigated whether organoids resemble the intestinal epithelium in their microRNA (miRNA) profiles. Total RNA samples were obtained from crypt and villus fractions in murine intestine and from cultured organoids. Microarray analysis showed that organoids largely resembled intestinal epithelial cells in their miRNA profiles. In silico prediction followed by qRT-PCR suggested that six genes are regulated by corresponding miRNAs along the crypt-villus axis, suggesting miRNA regulation of epithelial cell renewal in the intestine. However, such expression patterns of miRNAs and their target mRNAs were not reproduced during organoids maturation. This might be due to lack of luminal factors and endocrine, nervous, and immune systems in organoids and different cell populations between in vivo epithelium and organoids. Nevertheless, we propose that intestinal organoids provide a useful in vitro model to investigate miRNA expression in intestinal epithelial cells.
• Extra-adrenal glucocorticoids contribute to the postprandial increase of circulating leptin in mice

  Yuka Tomabechi, Takeshi Tsuruta, Shinichi Saito, Martin Wabitsch, Kei Sonoyama

  Journal of Cell Communication and Signaling 12 (2) 433 - 439 1873-9601 2018/06 [Refereed]
  
• 小腸腔内のセロトニンはL細胞においてセロトニン受容体HTR4を介してGLP-1分泌を抑制する

  奥村 元, 比良 徹, 園山 慶

  日本栄養・食糧学会大会講演要旨集 (公社)日本栄養・食糧学会 72回 187 - 187 2018/04
• Fructo-oligosaccharide-Induced Transient Increases in Cecal Immunoglobulin A Concentrations in Rats Are Associated with Mucosal Inflammation in Response to Increased Gut Permeability

  Tomomi Genda, Yuta Sasaki, Takashi Kondo, Shingo Hino, Naomichi Nishimura, Takamitsu Tsukahara, Kei Sonoyama, Tatsuya Morita

  JOURNAL OF NUTRITION 147 (10) 1900 - 1908 0022-3166 2017/10 [Refereed][Not invited]
   

  Background: The mechanism underlying transient increases in immunoglobulin (Ig) A concentrations in the cecal contents of rats fed fructo-oligosaccharide (FOS) is unclear. Objective: This study was designed to test whether increased IgA concentrations represent one aspect of the inflammatory response to increased permeability induced by FOS in the cecum. Methods: Seven-week-old male Wistar rats were fed a fiber-free semipurified diet (FFP) with or without supplemental FOS (60 g/kg diet) for 9 or 58 d [experiment (expt.) 1], 7 d (expt. 2), or 7 or 56 d (expt. 3). In addition to measuring IgA concentrations in cecal content, we assessed gut permeability, inflammatory responses (expt. 1), the number of IgA plasma cells in the cecal lamina propria, polymeric Ig receptor (pIgR) expression in the cecal mucosa (expt. 2), and the condition of the cecal mucus layer (expt. 3). Results: The cecal IgA concentration in the FOS-fed rats was 15-fold higher than that of the rats fed FFP for 9 d (P < 0.05). Gut permeability estimated by urinary chromium-EDTA excretion, bacterial translocation to mesenteric lymph nodes, myeloperoxidase activity, and expression of inflammatory cytokine genes in the cecal mucosa was greater in the FOS-fed rats than in the rats fed FFP for 9 d. These effects were not observed in the rats fed FOS for 58 d (expt. 1). Accompanying the higher cecal IgA concentration, pIgR protein and the number of IgA plasma cells in the cecal mucosa were higher in the FOS-fed rats than in the rats fed FFP for 7 d (expt. 2). Destruction of the mucus layer on the epithelial surface, as evidenced by Alcian blue staining in the cecal sections, was evident in the rats fed FOS for 7 d, but themucus layer appeared normal in the rats fed FOS for 56 d (expt. 3). Conclusions: These findings suggest that transient increases in cecal IgA concentrations induced by FOS in rats are associated with mucosal inflammation in response to increased gut permeability; these are presumably evoked by disruption of the cecal mucus barrier. The observed responses could contribute to the maturation of the gut immune system.
• Exosomes isolated from sera of mice fed Lactobacillus strains affect inflammatory cytokine production in macrophages in vitro

  Ayako Aoki-Yoshida, Shinichi Saito, Takeshi Tsuruta, Arisa Ohsumi, Hinako Tsunoda, Kei Sonoyama

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 489 (2) 248 - 254 0006-291X 2017/07 [Refereed][Not invited]
   

  Orally administered Lactobacillus strains, including L. plantarum No.14 and L rhamnosus GG, reportedly reduce inflammatory cytokine production in mice. The present study tested our idea that circulating exosomes mediate the action of Lactobacillus strains. The lipopolysaccharide-induced production of TNF-alpha and IL-6 in vitro was attenuated in peritoneal exudate cells (PECs) isolated from C57BL/6N mice that had been fed L. plantarum No.14. When PECs were cultured for 24 h with exosomes isolated from the serum of mice fed L plantarum No.14 or L. rhamnosus GG, accumulation of both TNF-alpha and of the corresponding mRNA was lowered. Growth in the presence of these exosomes also decreased the production of TNF-alpha and IL-6 by the murine macrophage cell line RAW264.7. In contrast, supplementation with exosome-depleted serum of mice fed L plantarum No.14 or L. rhamnosus GG failed to affect the production of TNF-alpha and IL-6 by RAW264.7 cells. When PECs and RAW264.7 cells were cultured for 24 h with PKH67-labeled exosomes isolated from murine serum, fluorescent signal was observed inside the cells, suggesting that these cells incorporate serum exosomes. We propose that the anti-inflammatory activity of orally administered L plantarum No.14 and L rhamnosus GG is mediated, at least in part, by circulating exosomes. (C) 2017 Elsevier Inc. All rights reserved.
• Organoids as an ex vivo model for studying the serotonin system in the murine small intestine and colon epithelium

  Takeshi Tsuruta, Shinichi Saito, Yosuke Osaki, Akihiro Hamada, Ayako Aoki-Yoshida, Kei Sonoyama

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 474 (1) 161 - 167 0006-291X 2016/05 [Refereed][Not invited]
   

  Intestinal organoids were recently established as an ex vivo model of the intestinal epithelium. The present study investigated the serotonin (5-hydroxytryptamine, 5-HT) system using organoids. Organoids from murine small intestinal and colonic crypts were successfully cultured. Reverse transcription-polymerase chain reaction (RT-PCR) analysis showed that small intestinal and colonic organoids express mRNAs encoding tryptophan hydroxylase-1 (TPH1) (the rate-limiting enzyme of 5-HT synthesis), serotonin reuptake transporter (SERT), 5-HT receptor (HTR)2A, HTR2B, and HTR4. SERT mRNA levels were significantly higher in the small intestine than in the colon in both the mucosal tissues and organoids, as estimated by quantitative real-time RT-PCR. Although the 5-HT concentration and levels of chromogranin A (CgA) (an enteroendocrine cell marker), TPH1, and HTR4 mRNAs were significantly higher in the colonic mucosa than the small intestinal mucosa, they were the same in small intestinal and colonic organoids. There were no significant differences in HTR2A and HTR2B mRNA levels between the small intestine and colon in either the mucosal tissues or organoids. Immunofluorescence staining showed that the number of CgA-positive cells in the colonic organoids appeared to increase upon culturing with acetate. Acetate supplementation significantly increased CgA, TPH1, and HTR4 mRNA levels in the colonic organoids. We propose that organoids are useful for investigating the 5-HT system in the intestinal epithelium, even though colonic organoids may require gut microbiota derived factors such as short-chain fatty acids. (C) 2016 Elsevier Inc. All rights reserved.
• Lactobacillus rhamnosus GG increases Toll-like receptor 3 gene expression in murine small intestine ex vivo and in vivo

  A. Aoki-Yoshida, S. Saito, S. Fukiya, R. Aoki, Y. Takayama, C. Suzuki, K. Sonoyama

  BENEFICIAL MICROBES 7 (3) 421 - 429 1876-2883 2016 [Refereed][Not invited]
   

  Administration of Lactobacillus rhamnosus GG (LGG) has been reported to be therapeutically effective against acute secretory diarrhoea resulting from the structural and functional intestinal mucosal lesions induced by rotavirus infection; however, the underlying mechanisms remain to be completely elucidated. Because Toll-like receptor 3 (TLR3) plays a key role in the innate immune responses following the recognition of rotavirus, the present study examined whether LGG influences TLR3 gene expression in murine small intestine ex vivo and in vivo. We employed cultured intestinal organoids derived from small intestinal crypts as an ex vivo tissue model. LGG supplementation increased TLR3 mRNA levels in the intestinal organoids, as estimated by quantitative real-time polymerase chain reaction. Likewise, single and 7-day consecutive daily administrations of LGG increased TLR3 mRNA levels in the small intestine of C57BL/6N mice. The mRNA levels of other TLRs were not substantially altered both ex vivo and in vivo. In addition, LGG supplementation increased the mRNA levels of an antiviral type 1 interferon, interferon-alpha (IFN-alpha), and a neutrophil chemokine, CXCL1, upon stimulation with a synthetic TLR3 ligand, poly(I:C) in the intestinal organoids. LGG administration did not alter IFN-alpha and CXCL1 mRNA levels in the small intestine in vivo. Supplementation of other bacterial strains, Bifidobacterium bifidum and Lactobacillus paracasei, failed to increase TLR3 and poly(I: C)-stimulated CXCL1 mRNA levels ex vivo. We propose that upregulation of TLR3 gene expression may play a pivotal role in the therapeutic efficacy of LGG against rotavirus-associated diarrhoea. In addition, we demonstrated that intestinal organoids may be a promising ex vivo tissue model for investigating host-pathogen interactions and the antiviral action of probiotics in the intestinal epithelium.
• Intestinal epithelial cells promote secretion of leptin and adiponectin in adipocytes

  Rino Ishihara, Yuki Mizuno, Akiho Miwa, Akihiro Hamada, Takeshi Tsuruta, Martin Wabitsch, Kei Sonoyama

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 458 (2) 362 - 368 0006-291X 2015/03 [Refereed][Not invited]
   

  Although leptin and adiponectin are the predominant adipokines, how their circulating levels are regulated is incompletely understood. The present study tested whether intestinal epithelial cells influence the expression and secretion of these adipokines by adipocytes. Leptin gene expression and secretion by cultured human primary adipocytes and Simpson-Golabi-Behmel Syndrome adipocytes increased upon coculture with human enterocytic Caco-2 cells or incubation in conditioned medium of Caco-2 cells. Although adiponectin secretion increased, its mRNA levels decreased. Tissue homogenate of the ileum (but not the jejunum, colon, or liver) of nonobese C57BL/6J mice also stimulated leptin and adiponectin secretion by cultured murine 3T3-L1 adipocytes. However, ileal homogenate of obese KK-Ay mice had no effect on leptin and adiponectin secretion. We propose that as yet unidentified humoral factors released from intestinal epithelial cells are involved in regulating circulating leptin and adiponectin levels. Decreased production of such factors may contribute to hyperphagia in KK-Ay mice. (C) 2015 Elsevier Inc. All rights reserved.
• Dual labeling with 5-bromo-2 '-deoxyuridine and 5-ethynyl-2 '-deoxyuridine for estimation of cell migration rate in the small intestinal epithelium

  Mami Asano, Tatsuro Yamamoto, Takeshi Tsuruta, Naomichi Nishimura, Kei Sonoyama

  DEVELOPMENT GROWTH & DIFFERENTIATION 57 (1) 68 - 73 0012-1592 2015/01 [Refereed][Not invited]
   

  Small intestinal epithelium is a self-renewing system in which the entire sequence of cell proliferation, differentiation, and removal is coupled to cell migration along the crypt-villus axis. We examined whether dual labeling with different thymidine analogues, 5-bromo-2'-deoxyuridine (BrdU) and 5-ethynyl-2'-deoxyuridine (EdU), can be used to estimate cell migration rates on the villi of small intestines in rats. Rats received a single intraperitoneal injection of BrdU and EdU within a time interval, and signals in tissue sections were examined by immunohistochemistry and the click reaction, respectively. We successfully observed BrdU- and EdU-positive cells on the epithelium with no cross-reaction. In addition, we observed an almost complete overlapping of BrdU- and EdU-positive cells in rats administered simultaneously with BrdU and EdU. By calculating the cell migration rate by dividing the distance between the median cell positions of the distribution of BrdU- and EdU-positive cells by the time between the injection of BrdU and EdU, we estimated approximately 9 and 5m/h for the cell migration rates on the villi in the jejunum and ileum, respectively. We propose that dual labeling with BrdU and EdU within a time interval, followed by detecting with immunohistochemistry and the click reaction, respectively, is useful to estimate accurately the cell migration rate in the intestinal epithelium in a single animal.
• Gastric emptying is involved in Lactobacillus colonisation in mouse stomach

  Yuraporn Sahasakul, Naoki Takemura, Kei Sonoyama

  BRITISH JOURNAL OF NUTRITION 112 (3) 408 - 415 0007-1145 2014/08 [Refereed][Not invited]
   

  Lactobacilli are indigenous microbes of the stomach of rodents, with much lower numbers being present in mice fed a purified diet than in those fed a non-purified diet. We postulated that gastric emptying (GE) is responsible for the different colonisation levels of lactobacilli and tested this hypothesis in the present study. BALB/cCr Slc mice were fed either a non-purified diet or a purified diet for 2 weeks. The number of gastric tissue-associated lactobacilli was lower in mice fed the purified diet than in those fed the non-purified diet. GE, estimated by measuring the food recovered from the stomach, was higher in mice fed the purified diet than in those fed the non-purified diet and correlated negatively with the number of lactobacilli. Mice fed the non-purified diet exhibited lower GE rates even when lactobacilli were eliminated by ampicillin administration through the drinking-water, suggesting that GE is the cause but not the consequence of different Lactobacillus colonisation levels. The plasma concentrations of acylated ghrelin, a gastric hormone that promotes GE, were higher in mice fed the purified diet than in those fed the non-purified diet. There was a negative correlation between GE and the number of lactobacilli in mice fed the non-purified diet, the purified diet, and the purified diet supplemented with sugarbeet fibre (200 g/kg diet) or carboxymethyl cellulose (40 g/kg diet). We propose that a higher GE rate contributes, at least in part, to lower gastric colonisation levels of lactobacilli in mice fed a purified diet.
• Adiponectin is partially associated with exosomes in mouse serum

  Worrawalan Phoonsawat, Ayako Aoki-Yoshida, Takeshi Tsuruta, Kei Sonoyama

  BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS 448 (3) 261 - 266 0006-291X 2014/06 [Refereed][Not invited]
   

  Exosomes are membrane vesicles 30-120 nm in diameter that are released by many cell types and carry a cargo of proteins, lipids, mRNA, and microRNA. Cultured adipocytes reportedly release exosomes that may play a role in cell-to-cell communication during the development of metabolic diseases. However, the characteristics and function of exosomes released from adipocytes in vivo remain to be elucidated. Clearly, adipocyte-derived exosomes could exist in the circulation and may be associated with adipocyte-specific proteins such as adipocytokines. We isolated exosomes from serum of mice by differential centrifugation and analyzed adiponectin, leptin, and resistin in the exosome fraction. Western blotting detected adiponectin but no leptin and only trace amounts of resistin in the exosome fraction. The adiponectin signal in the exosome fraction was decreased by proteinase K treatment and completely quenched by a combination of proteinase K and Triton X-100. Quantitative ELISA showed that the exosome fraction contains considerable amounts of adiponectin, but not leptin or resistin. The concentration of adiponectin in the serum and the ratio of adiponectin to total protein in the exosome fraction were lower in obese mice than in lean mice. These results suggest that a portion of adiponectin exists as a transmembrane protein in the exosomes in mouse serum. We propose adiponectin as a marker of exosomes released from adipocytes in vivo. (C) 2014 Elsevier Inc. All rights reserved.
• Low-Methoxyl Pectin Stimulates Small Intestinal Mucin Secretion Irrespective of Goblet Cell Proliferation and Is Characterized by Jejunum Muc2 Upregulation in Rats

  Shingo Hino, Kei Sonoyama, Hiroyuki Bito, Hirokazu Kawagishi, Seiichiro Aoe, Tatsuya Morita

  JOURNAL OF NUTRITION 143 (1) 34 - 40 0022-3166 2013/01 [Refereed][Not invited]
   

  Generally, soluble fibers increase small intestinal mucin secretion by increasing the number of goblet cells in a viscosity-dependent manner. The present study aimed to examine the mechanism by which low-methoxyl pectin (LPC) affects mucin secretion in the small intestine. First, diets containing 50 g/kg of low-viscosity fiber (LPC, gum arabic, guar gum, low-molecular konjac mannan, arabinogalactan, sodium alginate) or high-molecular konjac mannan (KMH) were fed to Wistar rats for 10 d. Luminal mucin was greater in the LPC and KMH groups than in the fiber-free control group, but only the KMH group had more goblet cells in the ileum compared with the other groups. Next, Sprague-Dawley rats were fed LPC, KMH, or high-methoxyl pectin (HPC) diets 150 g/kg) for 10 d. The KMH and LPC groups, but not the HPC group, had greater luminal mucin than the control group, whereas jejunum Muc2 expression was higher only in the LPC group. Sprague-Dawley rats fed the LPC diet for 1 or 3 d had greater luminal mucin and jejunum Muc2 expression than those fed the control diet. In vitro studies using HT-29MTX cells showed that, of the various fibers studied, only LPC and HPC affected mucin secretion. Finally, Wistar rats were fed the LPC diet with or without neomycin in drinking water for 10 d; neomycin treatment did not compromise the effect of LPC on mucin secretion. We conclude that LPC does not affect the number of goblet cells but can interact directly with the epithelium and stimulate small intestinal mucin secretion. J. Nutr. 143: 34-40, 2013.
• KK/Ta Mice Administered Lactobacillus plantarum Strain No. 14 Have Lower Adiposity and Higher Insulin Sensitivity.

  Okubo T, Takemura N, Yoshida A, Sonoyama K

  Bioscience of microbiota, food and health 32 (3) 93 - 100 2186-6953 2013 [Refereed][Not invited]
  
• Different Impacts of Purified and Nonpurified Diets on Microbiota and Toll-Like Receptors in the Mouse Stomach

  Yuraporn Sahasakul, Naoki Takemura, Kei Sonoyama

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 76 (9) 1728 - 1732 0916-8451 2012/09 [Refereed][Not invited]
   

  We compared the colonization of lactobacilli in the stomachs of mice fed nonpurified and purified diets and examined to determine whether the expression of Toll-like receptor 2, which is involved in the recognition of lactobacilli, is influenced by diet. Female BALB/c mice were fed a nonpurified or a purified diet for 2 weeks. Conventional cultivation and cultivation-independent molecular biological analysis based on the 16S rRNA gene sequence revealed that the number of lactobacilli associated with the gastric tissue was significantly higher in the mice fed the nonpurified diet than in those fed the purified diet. Sequencing analysis indicated that L. gasseri and L. johnsonii were predominant Lactobacillus species associated with the gastric tissue of the mice fed the nonpurified diet. The mRNA levels of Toll-like receptor 2, but not of 9, in the gastric tissue were significantly higher in the mice fed the purified diet than in those fed the nonpurified diet. We propose that nonpurified and purified diets have different impacts on gastric microbiota, which can in turn influence the expression of Toll-like receptor 2 in the stomach.
• Small Intestinal Goblet Cell Proliferation Induced by Ingestion of Soluble and Insoluble Dietary Fiber Is Characterized by An Increase in Sialylated Mucins in Rats

  Shingo Hino, Naoki Takemura, Kei Sonoyama, Akio Morita, Hirokazu Kawagishi, Seiichiro Aoe, Tatsuya Morita

  JOURNAL OF NUTRITION 142 (8) 1429 - 1436 0022-3166 2012/08 [Refereed][Not invited]
   

  The study aimed to examine the effects of insoluble and soluble fibers on mucin sialylation and sulfation in the small intestine. First, diets containing soluble [konjac mannan (KM), psyllium, or guar gum; 50 g/kg) or insoluble (polystyrene foam, wheat bran, or cornhusk; 80 g/kg) fiber were fed to rats for 13 d. The fiber-fed groups had more goblet cells in the ileum than the fiber-free control group. High-iron diamine/alcian blue staining showed more sialylated mucin-producing cells in the fiber-fed groups than in the control, whereas sulfated mucin-producing cells were fewer (insoluble fibers) or unchanged (soluble fibers). Second, feeding KM (50 g/kg) and beet fiber (BF) (80 g/kg) diets for 7 d yielded a higher ileum Siat4C expression than the control, but Gal3ST2 and Gal3ST4 expression was comparable. Luminal mucin content correlated with sialic acid (r = 0.96; P < 0.001) or sulfate (r = 0.62; P < 0.01), but the slope of the slake acid-derived equation was greater than that of the sulfate-derived equation, indicating a preferred increase in sialylated mucins. Third, rats were fed the control diet for 10 d while receiving antibiotic treatment. Analysis of the luminal mucin showed that sialylated mucins were more vulnerable to bacterial degradation than sulfated mucins. Finally, a study of bromo-deoxyuridine incorporation in rats fed a BF diet indicated that goblet cell proliferation accompanied by increased sialylated mucin appeared to be related to accelerated ileal epithelial cell migration. We conclude that intestinal goblet cell responses to insoluble and soluble fibers are characterized by increases in sialylated mucin production. J. Nutr. 142: 1429-1436, 2012.
• Reduced Expression of Polymeric Immunoglobulin Receptors in the Intestine of Young Rats Fed a Fiber-free Diet.

  Nakamura Y, Sonoyama K

  Bioscience of microbiota, food and health 31 (3) 51 - 58 2186-6953 2012 [Refereed][Not invited]
  
• Cell Wall Polysaccharides of Candida albicans Induce Mast Cell Degranulation in the Gut.

  Sakurai A, Yamaguchi N, Sonoyama K

  Bioscience of microbiota, food and health 31 (3) 67 - 70 2186-6953 2012 [Refereed][Not invited]
  
• Gut Colonization by Candida albicans Inhibits the Induction of Humoral Immune Tolerance to Dietary Antigen in BALB/c Mice.

  Sugita R, Hata E, Miki A, Andoh R, Umeda C, Takemura N, Sonoyama K

  Bioscience of microbiota, food and health 31 (4) 77 - 84 2186-6953 2012 [Refereed][Not invited]
  
• Persistence of orally administered lactobacillus strains in the gut of infant mice.

  Ozawa K, Fujiwara R, Watanabe K, Sonoyama K

  Bioscience of microbiota, food and health 31 (4) 85 - 91 2186-6953 2012 [Refereed][Not invited]
  
• Degree of Polymerization of Inulin-Type Fructans Differentially Affects Number of Lactic Acid Bacteria, Intestinal Immune Functions, and Immunoglobulin A Secretion in the Rat Cecum

  Hiroyuki Ito, Naoki Takemura, Kei Sonoyama, Hirokazu Kawagishi, David L. Topping, Michael A. Conlon, Tatsuya Morita

  JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 59 (10) 5771 - 5778 0021-8561 2011/05 [Refereed][Not invited]
   

  This study examined the role of degree of polymerization (DP) of inulin-fructans in modulating the interaction between lactic acid bacteria and IgA cecal secretion. Rats were fed a control diet or a diet containing one of the fructans with different DP. Consuming fructans increased the cecal IgA concentrations in the order DP4 > DP8 > DP16. Cecal lactobacilli counts were higher in DP4, DP8, and DP16, whereas bifidobacteria were higher in DP8, DP16, and DP23. Cecal IgA concentrations were correlated with cecal lactobacilli counts (P < 0.01). DP4, DP8, and DP16, but not DP23, significantly increased IgA-producing plasma cells In the cecal mucosa. IFN-gamma and IL-10 production in the cecal CD4(+) T cells was enhanced solely in DP4. The results show that fructans with lower DP enhance cecal IgA secretion and increase the plasma cells and suggest that the increased lactobacilli may contribute to the stimulation of cecal IgA secretion.
• Gut colonization by Candida albicans aggravates inflammation in the gut and extra-gut tissues in mice

  Kei Sonoyama, Atsuko Miki, Ryusuke Sugita, Haruka Goto, Mayumi Nakata, Natsu Yamaguchi

  MEDICAL MYCOLOGY 49 (3) 237 - 247 1369-3786 2011/04 [Refereed][Not invited]
   

  We examined whether Candida albicans gut colonization aggravates immune diseases in mice. Chronic and latent C. albicans gut colonization was established by the intragastric inoculation of C. albicans in mice fed as part of a purified diet. Allergic diarrhea was induced by repetitive intragastric administration of ovalbumin in sensitized BALB/c mice. Contact hypersensitivity was evaluated by measuring ear swelling after topical application of 2, 4-dinitrofluorobenzene in NC/Nga mice. Arthritis was induced by intradermal injection of bovine type-II collagen emulsified with complete Freund's adjuvant in DBA/1J mice. C. albicans gut colonization increased the incidence of allergic diarrhea, which was accompanied by gut hyperpermeability, as well as increased infiltration of inflammatory cells in the colon. Contact hypersensitivity was also exacerbated by C. albicans gut colonization, as demonstrated by increased swelling, myeloperoxidase activity, and proinflammatory cytokines in ear auricles. Furthermore, C. albicans gut colonization promoted limb joint inflammation in collagen-induced arthritis, in an animal model of rheumatoid arthritis. These findings suggest that C. albicans gut colonization in mice aggravates inflammation in allergic and autoimmune diseases, not only in the gut but also in the extra-gut tissues and underscores the necessity of investigating the pathogenic role of C. albicans gut colonization in immune diseases in humans.
• Effects of Fructo-Oligosaccharide on DSS-Induced Colitis Differ in Mice Fed Nonpurified and Purified Diets

  Haruka Goto, Naoki Takemura, Toru Ogasawara, Naho Sasajima, Jun Watanabe, Hiroyuki Ito, Tatsuya Morita, Kei Sonoyama

  JOURNAL OF NUTRITION 140 (12) 2121 - 2127 0022-3166 2010/12 [Refereed][Not invited]
   

  We investigated whether feeding a purified compared with nonpurified diet supplemented with or without fructooligosaccharide (FOS 50 g/kg diet) altered the response of C57BL/6 mice to DSS induced diarrhea In Expt 1 we examined disease severity in mice receiving DSS (2% in drinking water) for 5 d In Expt 2 we measured cecal organic acid concentrations and fecal water-holding capacity (WHC) In Expts 3 and 4 we tested whether polycarbophil calcium (PC) a water-absorbing polymer altered fecal WHC and disease severity FOS exacerbated diarrhea and weight loss in mice fed the purified diet and reduced fecal bleeding in mice fed the nonpurified diet (P < 0 05) Without DSS administration cecal acetate and butyrate concentrations were higher in mice fed the nonpurified diet than in mice fed the purified diet (P < 0 05) Fecal WHC was higher in mice fed the nonpurified diet than in mice fed the purified diet (P < 0 05) One day after starting DSS administration cecal succinate concentrations were higher in mice fed the FOS-supplemented purified diet than in mice fed the other 3 diets whereas SCFA concentrations were higher in mice fed the nonpurified diet than in mice fed the purified diet (P< 0 05) PC supplementation increased fecal WHC and prevented FOS exacerbation of diarrhea in mice fed the purified diet (P < 0 05) We conclude that the effects of FOS on DSS-induced diarrhea differ in mice fed the purified and nonpurified diets The protective effect of nonpurified diet was associated with increased production of organic acids and WHC in the intestinal contents J Nutr 140 2121-2127 2010
• Inulin Prolongs Survival of Intragastrically Administered Lactobacillus plantarum No. 14 in the Gut of Mice Fed A High-Fat Diet

  Naoki Takemura, Masahiro Hagio, Satoshi Ishizuka, Hiroyuki Ito, Tatsuya Morita, Kei Sonoyama

  JOURNAL OF NUTRITION 140 (11) 1963 - 1969 0022-3166 2010/11 [Refereed][Not invited]
   

  We tested whether a high-fat diet (HFD) impairs the survival of probiotics in mice. In Expt. 1, after feeding either a HFD (62.7% energy) or a normal-fat diet (NFD; 11.1% energy) for 2 d, C57BL16 mice were i.g. administered Lactobacillus plantarum No. 14. Fecal recovery of viable L. plantarum was significantly decreased 99% by the HFD compared with the NFD. Total bile acid concentrations in the small intestine and cecum were significantly higher 11.5- and 2.2-fold of NFD, respectively) in mice fed HFD than in those fed NFD. Cholic acid and deoxycholic acid significantly reduced the viability of L. plantarum No. 14 in culture experiments. In Expt. 2, after feeding HFD for 2 d, simultaneous administration of inulin 110 mg) with L. plantarum No. 14 significantly increased (100-fold of that without inulin) the fecal recovery of viable L. plantarum. Inulin administration did not alter intestinal bile acid concentrations. In Expt. 3, after feeding HFD for 2 d, mice were i.g. administered either inulin (10 mg) or vehicle and, after 6 h, cecal contents were subjected to culture experiments. Growth of L. plantarum No. 14 was significantly higher in the cecal contents of inulin-administered mice than vehicle-administered mice. Inulin supplementation to cecal contents of vehicle-administered mice significantly enhanced the growth of L. plantarum No. 14. We propose that HFD impairs the survival of probiotics in the gut due to increased bile acid stress and that simultaneous administration of inulin prolongs the survival of probiotics in mice fed HFD. J. Nutr. 140: 1963-1969, 2010.
• 2,4-Dinitrofluorobenzene-Induced Contact Hypersensitivity Response in NC/Nga Mice Fed Fructo-Oligosaccharide

  Reiko Fujiwara, Naho Sasajima, Naoki Takemura, Keisuke Ozawa, Yuki Nagasaka, Takuma Okubo, Yuraporn Sahasakul, Jun Watanabe, Kei Sonoyama

  JOURNAL OF NUTRITIONAL SCIENCE AND VITAMINOLOGY 56 (4) 260 - 265 0301-4800 2010/08 [Refereed][Not invited]
   

  Strategies to manipulate gut microbiota in infancy have been considered to prevent the development of allergic diseases later in life. We previously demonstrated that maternal dietary supplementation with fructo-oligosaccharide (FOS) during pregnancy and lactation modulated the composition of gut microbiota and diminished the severity of spontaneously developing atopic dermatitis-like skin lesions in the offspring of NC/Nga mice. The present study tested whether dietary FOS affects contact hypersensitivity (CHS), another model for allergic skin disease, in NC/Nga mice. In experiment 1, 5-wk-old female NC/Nga mice were fed diets either with or without FOS supplementation for 3 wk and then received 2,4-dinitrofluorobenzene (DNFB) on the ear auricle 5 times at 7-d intervals. PUS supplementation reduced CHS response as demonstrated by ear swelling. Quantitative RT-PCR analysis showed that mRNA levels for interleukin (IL)-10, IL-12p40, and IL-17 in the lesional ear skin were significantly lower in mice fed FOS. In experiment 2, female NC/Nga mice were fed diets either with or without PUS during pregnancy and lactation. After weaning. offspring were led the diets supplemented with or without FOS. Three weeks after weaning, offspring received DNFB on the ear auricle 4 times at 7-d intervals. Although PUS supplementation after weaning reduced ear swelling, maternal FOS consumption was ineffective in offspring. The present data suggest that dietary FOS reduces CHS while maternal FOS consumption is ineffective in offspring of DNFB-treated NC/Nga mice.
• Lactobacillus plantarum strain No. 14 reduces adipocyte size in mice fed high-fat diet

  Naoki Takemura, Takuma Okubo, Kei Sonoyama

  EXPERIMENTAL BIOLOGY AND MEDICINE 235 (7) 849 - 856 1535-3702 2010/07 [Refereed][Not invited]
   

  Because gut microbiota has recently attracted much attention as an environmental factor involved in the development of obesity, probiotics may be useful in preventing and/or improving obesity and its related disorders. The present study aimed to investigate the effects of Lactobacillus plantarum strain No. 14 (LP14), a bacterial strain reported to decrease body fat percentage in healthy volunteers, on adipocyte size in mice. Female C57BL/6 mice were fed either normal- or high-fat diet and administered intragastrically with LP14 (1 x 10(8) colony-forming units/mouse) or vehicle daily for 11 weeks. High dietary fat intake increased body weight gain, white adipose tissue weight, mean adipocyte size and serum total cholesterol and leptin concentrations, and decreased serum adiponectin concentration. In mice fed the high-fat diet, LP14 administration significantly reduced the mean adipocyte size and tended to reduce the white adipose tissue weight and serum total cholesterol and leptin concentrations as compared with the vehicle-administered mice. All mice had undetectable serum levels of conjugated linoleic acids that reportedly exert antiobesity action. In a separate experiment, LP14 ingestion had no influence on serum triacylglycerol accumulation following olive oil administration in Triton WR1339-treated mice, suggesting that dietary fat absorption is unaffected by LP14. In conclusion, we propose that LP14 may exert a beneficial effect on the onset of diet-induced obesity by reducing the cell size of white adipose tissues, and it seems unlikely that previously reported mechanisms for other bacterial strains are involved in the action of LP14.
• Inulin-Type Fructans Stimulated the Growth of Exogenously Administered Lactobacillus plantarum No. 14 in the Mouse Gastrointestinal Tract

  Naoki Takemura, Kesuke Ozawa, Naoto Kimura, Jun Watanabe, Kei Sonoyama

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 74 (2) 375 - 381 0916-8451 2010/02 [Refereed][Not invited]
   

  This stud), examined the effects of the inulin-type fructans known as fructo-oligosaccharide (FOS) and inulin on the growth and persistence or Lactobacillus plantarum No. 14 (LP14) in the gut. Cultivation in medium that contained either FOS or inulin revealed that both fructans supported the growth of LP14. Fecal recover), of viable L. plantarum at 24 and 30 h following administration of LP14 were highest in mice red inulin, followed by mice red FOS and the control diet. However, neither of the fructans prolonged the appearance of LP14 in the gut. Cultivation in fecal extract medium prepared from mice red each of the test diets showed higher growth of LP14 in the mice red fructans. Histological analysis following fluorescein-stained LP14 administration showed that LP14 was largely localized to the luminal contents. These results suggest that inulin-type fructans support the growth of LP14 in the luminal contents only during their passage through the gut.
• Administration of Antibiotics during Infancy Promoted the Development of Atopic Dermatitis-Like Skin Lesions in NC/Nga Mice

  Jun Watanabe, Reiko Fujiwara, Naho Sasajima, Susumu Ito, Kei Sonoyama

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 74 (2) 358 - 363 0916-8451 2010/02 [Refereed][Not invited]
   

  This study aimed to determine whether oral antibiotic administration during infancy is associated with the spontaneous development of atopic dermatitis-like skin lesions by modulating intestinal microbiota. Female NC/Nga mice at 3 weeks of age were orally administered kanamycin or polymyxin B. Clinical symptoms, scratching behavior, and serum antibody levels were evaluated. Changes in intestinal microbiota were determined by culture-independent analysis and cultural analysis. The kanamycin-treated mice showed higher clinical scores and scratching frequency than the control mice. IgE levels were significantly higher in the kanamycin-treated mice than in the control mice. Transient changes in intestinal microbiota were observed under kanamycin treatment. Polymyxin B treatment failed to affect scratching behavior. These results suggest that oral administration of kanamycin during infancy promoted the development of atopic dermatitis-like skin lesions in NC/Nga mice and was associated with a transient change in intestinal microbiota.
• Maternal consumption of fructo-oligosaccharide diminishes the severity of skin inflammation in offspring of NC/Nga mice

  Reiko Fujiwara, Naoki Takemura, Jun Watanabe, Kei Sonoyama

  BRITISH JOURNAL OF NUTRITION 103 (4) 530 - 538 0007-1145 2010/02 [Refereed][Not invited]
   

  Strategies to manipulate the gut microbiota in infancy have been considered to prevent the development of allergic diseases later in life. We aimed to elucidate the effects of maternal dietary supplementation with a prebiotic oligosaccharide on gut microbiota and spontaneously developing atopic dermatitis-like skin lesions in the offspring of NC/Nga mice. Female NC/Nga mice were fed diets either with or without fructooligosaccharide supplementation during pregnancy and lactation. After weaning, offspring were fed the diets supplemented with or without fructo-oligosaccharide for I I weeks in an air-uncontrolled conventional room. Changes in gut microbiota were assessed by denaturing gradient gel electrophoresis of the PCR-amplified 16S rRNA gene. Skin lesions were evaluated by a clinical score and scratching behaviour. Serum antibody levels were measured by ELISA, and expression levels of cytokines and chemokines in lesional tissue were evaluated by quantitative RT-PCR. Maternal supplementation with fructo-oligosaccharide modulated the gut microbiota in sucklings. Although maternal supplementation with fructo-oligosaccharide suppressed the increase in clinical skin severity score and scratching behaviour in offspring, dietary fructooligosaccharide after weaning was less effective. The diminution of skin lesions was accompanied by lower serum concentrations of total IgGl and lower expression levels of TNF-alpha in the lesional tissue. These data suggest that maternal consumption of fructo-oligosaccharide diminishes the severity of atopic dermatitis-like skin lesions in the offspring of NC/Nga mice.
• Role of intestinal Bifidobacterium pseudolongum in dietary fructo-oligosaccharide inhibition of 2,4-dinitrofluorobenzene-induced contact hypersensitivity in mice

  Naho Sasajima, Toru Ogasawara, Naoki Takemura, Reiko Fujiwara, Jun Watanabe, Kei Sonoyama

  BRITISH JOURNAL OF NUTRITION 103 (4) 539 - 548 0007-1145 2010/02 [Refereed][Not invited]
   

  Strategies to manipulate the gut microbiota have been explored for preventing allergy development. We previously showed that dietary supplenlentation with fructo-oligosaccharide (FOS) reduced 2, 4-dinitrofluorobenzene (DNFB)-induced contact hypersensitivity (CHS) in BALB/c mice. Because the CHS response was negatively correlated with the number of faecal bifidobacteria, particularly Bifidobacterium pseudolongum, the present Study aimed to examine whether oral administration of B. pseudolongum affects CHS response. Viable B. pseudolongum was successfully isolated from mouse faeces. Female BALB/c mice were fed a synthetic diet with or without FOS supplementation, and B. pseudolongum (2 X 10(7) cells) was administered daily throughout the experimental period. Two weeks after starting the test diets, mice received DNFB on the ear auricle twice at 7-d intervals. Conventional cultivation and molecular biological analyses based on 16S rRNA gene sequences showed that administration of FOS and B. pseudolongum resulted in higher excretion of viable bifidobacteria, mainly B. psendolongum. Although dietary FOS reduced the CHS response as demonstrated by ear swelling, B. pseudolongum administration resulted in a reduction in the initial phase only of the CHS response. B. pseudolongum administration increased hapten-specific IgGI, while dietary FOS decreased IgG2a in sera. Administration of FOS and B. pseudolongum decreased interferon-gamma production and increased IL-10 production in cervical lymph node cells restimulated with hapten in vitro. We conclude that B. pseudolongum proliferation in the intestinal tract is partially responsible for the reduction in DNFB-induced CHS response by dietary Supplementation with FOS in mice, which may be mediated by the modulation of antigen-induced cytokine production.
• Comparison of gut microbiota and allergic reactions in BALB/c mice fed different cultivars of rice

  Kei Sonoyama, Toru Ogasawara, Haruka Goto, Tomoyo Yoshida, Naoki Takemura, Reiko Fujiwara, Jun Watanabe, Hiroyuki Ito, Tatsuya Morita, Yoshinari Tokunaga, Tetsuji Yanagihara

  BRITISH JOURNAL OF NUTRITION 103 (2) 218 - 226 0007-1145 2010/01 [Refereed][Not invited]
   

  Our preliminary clinical trial showed that consumption of cooked rice of a Japanese common cultivar Yukihikari improved atopic dermatitis associated with a suspected rice allergy, although the underlying mechanisms remain unclear. We hypothesised that the ameliorating effect of Yukihikari on atopic dermatitis is associated with the gut microbiota. BALB/c mice were fed a synthetic diet supplemented with uncooked and polished white rice powder prepared from one of four different cultivars: Yukihikari, rice A (common rice), rice B (brewery rice) and rice C (waxy rice). Denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments showed that the composition of faecal microbiota was different between mice fed Yukihikari and those fed rice A. Analysis of the 16S rRNA clone library and species-specific real-time PCR showed that the abundance of Akkermansia muciniphila, a mucin degrader, tended to be lower in mice fed Yukihikari. The incidence of allergic diarrhoea induced by oral administration of ovalbumin in systemically immunised mice was lower in mice fed Yukihikari, albeit with no difference in serum antibodies specific to ovalbumin. In a separate experiment, serum antibody levels specific to orally administered ovalbumin were lower in mice fed Yukihikari. Additionally, the translocation of horseradish peroxidase in isolated segments of ileum and colon tended to be lower in mice fed Yukihikari, suggesting a reduction in gut permeability in mice fed Yukihikari. These data indicate that changes in the gut microbiota of mice fed Yukihikari could be advantageous in the prevention of food allergy.
• Non-effect of maternal consumption of fructo-oligosaccharide on oral tolerance induction and gut mucosal barrier in offspring of BALB/c mice

  吉田知世, 後藤悠, 武村直紀, 藤原麗子, 園山慶

  Journal of Japanese Association for Dietary Fiber Research 日本食物繊維学会 14 (1) 23 - 32 1349-5437 2010 [Refereed][Not invited]
  
• Response of Gut Microbiota to Fasting and Hibernation in Syrian Hamsters

  Kei Sonoyama, Reiko Fujiwara, Naoki Takemura, Toru Ogasawara, Jun Watanabe, Hiroyuki Ito, Tatsuya Morita

  APPLIED AND ENVIRONMENTAL MICROBIOLOGY 75 (20) 6451 - 6456 0099-2240 2009/10 [Refereed][Not invited]
   

  Although hibernating mammals wake occasionally to eat during torpor, this period represents a state of fasting. Fasting is known to alter the gut microbiota in nonhibernating mammals; therefore, hibernation may also affect the gut microbiota. However, there are few reports of gut microbiota in hibernating mammals. The present study aimed to compare the gut microbiota in hibernating torpid Syrian hamsters with that in active counterparts by using culture-independent analyses. Hamsters were allocated to either torpid, fed active, or fasted active groups. Hibernation was successfully induced by maintaining darkness at 4 degrees C. Flow cytometry analysis of cecal bacteria showed that 96-h fasting reduced the total gut bacteria. This period of fasting also reduced the concentrations of short chain fatty acids in the cecal contents. In contrast, total bacterial numbers and concentrations of short chain fatty acids were unaffected by hibernation. Denaturing gradient gel electrophoresis of PCR-amplified 16S rRNA gene fragments indicated that fasting and hibernation modulated the cecal microbiota. Analysis of 16S rRNA clone library and species-specific real-time quantitative PCR showed that the class Clostridia predominated in both active and torpid hamsters and that populations of Akkermansia muciniphila, a mucin degrader, were increased by fasting but not by hibernation. From these results, we conclude that the gut microbiota responds differently to fasting and hibernation in Syrian hamsters.
• Soluble Fiber Viscosity Affects Both Goblet Cell Number and Small Intestine Mucin Secretion in Rats

  Hiroyuki Ito, Mitsuru Satsukawa, Eiko Arai, Kimio Sugiyama, Kei Sonoyama, Shuhachi Kiriyama, Tatsuya Morita

  JOURNAL OF NUTRITION 139 (9) 1640 - 1647 0022-3166 2009/09 [Refereed][Not invited]
   

  We examined the role of soluble fiber viscosity in small intestinal mucin secretion. Viscosities were defined as the area under the viscosity curve (VAUC). Rats were fed a control diet or diets containing konjac mannan (KM) [low, medium, or high molecular weight (LKM, MKM, HKM), respectively] at 50 g/kg diet for 10 d. Luminal mucin content and goblet cell number increased in proportion to the molecular weight of KM. Such effects with the HKM diet were nullified by the concurrent ingestion of 2 g cellulase/kg diet. Diet containing LKM, MKM, HKM, guar gums (high or low molecular weight; HGG, LGG), psyllium (PS), or pectin (PC) at 50 g/kg was fed to rats. Fibers with higher VAUC (MKM, HKM, HGG, and PS) increased goblet cell numbers, but not those with lower VAUC (LKM, LGG, and PC). Luminal mucins were greater in rats fed HKM, PC, and PS diets. Goblet cell numbers and VAUC were correlated (r = 0.98; P < 0.01). In rats fed the HKM diet, ileal Muc2 gene expression was not affected, but that of Muc3 was lower than in those fed the control diet, indicating that the increase in luminal mucins after ingestion of HKM diet occurred independently of enhanced Muc gene expression. An incorporation study of 5'-bromo-deoxyuridine (BrdU) showed the position of the uppermost-BrdU labeled cell along the villi was higher in rats fed the HKM diet than in those fed the control diet. The results suggest that soluble fibers, except PC, upregulate baseline secretion of luminal mucins by increasing goblet cell numbers in proportion to fiber VAUC. J. Nutr. 139:1640-1647, 2009.
• Elimination mechanism of Candida albicans in the colon of BALB/c mice by dietary fructo-oligosaccharide

  Dynamic Biochemistry, Process Biotechnology and Molecular Biology 3 85 - 89 2009 [Refereed][Not invited]
  
• Consumption of fructo-oligosaccharide reduces 2,4-dinitrofluorobenzene-induced contact hypersensitivity in mice

  Jun Watanabe, Naho Sasajima, Akiko Aramaki, Kei Sonoyama

  BRITISH JOURNAL OF NUTRITION 100 (2) 339 - 346 0007-1145 2008/08 [Refereed][Not invited]
   

  Strategies to manipulate the intestinal microbiota have been considered to promote immune health. The aim of the present study was to examine whether fructo-oligosaccharide, a typical prebiotic, could suppress antigen-specific skin inflammation by favourably changing the population of intestinal microbiota. Female BALB/c mice were fed a synthetic diet with or Without fructo-oligosaccharide supplementation for 3 weeks and were then epicutaneously immunised with 2,4-dinitrofluorobenzene. Afterwards, mice continued to receive their respective diets. At 5 d after immunisation, the mice were ear challenged with the hapten. Ear swelling after the challenge was significantly reduced in the mice fed the diet supplemented with fructo-oligosaccharide than in mice fed the control diet. To characterise the change in the intestinal microbiota, DNA samples isolated from fresh faeces were subjected to PCR-clenaturing gradient gel electrophoresis and real-time PCR based on 16S rDNA gene sequences. Dietary fructo-oligosaccharide altered the composition of intestinal microbiota. The numbers of bifidobacteria, but not lactobacilli, were significantly higher in mice fed the fructo-oligosaccharide-supplemented diet than in mice fed the control diet. Ear swelling was negatively correlated with the numbers of bifidobacteria in the faeces. Sequence analysis revealed that Bifidobacterium pseudolongum was the most predominant bifidobacteria in the intestine of mice fed the fructo-oligosaccharide-supplemented diet. These results suggest that consumption of fructo-oligosaccharide reduces contact hypersensitivity, which is associated with proliferation of B. pseudolongum in the intestinal tract of mice.
• Dietary alpha-linked galacto-oligosaccharide suppresses ovalbumin-induced allergic peritonitis in BALB/c mice

  Sooksan Pirapatdit, Eriko Kishino, Koki Fujita, Hiroyuki Hashimoto, Shigeharu Mori, Sanshiro Saito, Toshiro Sato, Jun Watanabe, Kei Sonoyama

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 72 (7) 1901 - 1907 0916-8451 2008/07 [Refereed][Not invited]
   

  To determine whether alpha-linked galacto-oligosaccharide (alpha-GOS) prevents allergic peritonitis, BALB/c mice were fed a synthetic diet with and without alpha-GOS supplementation for 7 d, and were then subcutaneously immunized with ovalbumin on days 0 and 7. The mice were challenged by intraperitoneal injection with ovalbumin on day 14, followed by peritoneal lavage on day 15. The total number of peritoneal exudate cells was significantly lower in the mice fed the alpha-GOS diet than in those fed the control diet. Peritoneal lavage fluid from mice fed the alpha-GOS diet not only had less potency to attract peripheral blood leukocytes and peritoneal exudate cells ex vivo, but also had lower concentrations of monocyte chemoattractant protein-1 (MCP-1) and eotaxin. Preincubation of the cells with alpha-GOS failed to affect the migration to peritoneal lavage fluid. We propose that dietary alpha-GOS reduces cell infiltration in allergic peritonitis by reducing antigen-induced elicitation of MCP-1 and eotaxin in mice.
• Assessing changes in composition of intestinal microbiota in neonatal BALB/c mice through cluster analysis of molecular markers

  Reiko Fujiwara, Jun Watanabe, Kei Sonoyama

  BRITISH JOURNAL OF NUTRITION 99 (6) 1174 - 1177 0007-1145 2008/06 [Refereed][Not invited]
   

  The present study introduced a molecular biological approach to demonstrate changes in the composition of intestinal microbiota in neonatal mice. Female BALB/c mice were fed either a control diet or a. diet supplemented with fructo-oligosaccharide (FOS) at 50 g/kg diet, and then mated to male mice. A cultivation-independent approach, denaturing gradient gel electrophoresis (DGGE) of the PCR-amplified 16S rRNA gene, was performed to characterise changes in intestinal microbial populations in pups at 0, 7, 14 and 21 d old and their dams. Comparisons of DGGE profiles were performed using the Dice similarity coefficient and the unweighted pair group method with arithmetic mean (UPGMA) cluster analysis based on numbers, positions and intensities of bands. DGGE profiles differed between dams fed control and FOS-supplemented diets. Although profiles in pups on the day of birth showed a high similarity with dams, profiles in 7-d-old pups differed from dams and showed high similarity to litter-mates. In 14- and 21-d-old pups, profiles again showed high similarity with dams. DGGE profiles in pups were divided into two large clusters of control and FOS-supplemented diet groups in the range of 0- to 21-d-old, suggesting modulation of intestinal microbiota in infants by manipulation of microbiota in dams. The present study shows a useful technique for demonstrating changes in intestinal microbiota and provides a mouse model for modulation of intestinal microbiota in neonatal life.
• Gastrointestinal Candida colonisation promotes sensitisation against food antigens by affecting the mucosal barrier in mice

  N Yamaguchi, R Sugita, A Miki, N Takemura, J Kawabata, J Watanabe, K Sonoyama

  GUT 55 (7) 954 - 960 0017-5749 2006/07 [Refereed][Not invited]
   

  Backgrounds and aims: Controversy still exists as to whether gastrointestinal colonisation by Candida albicans contributes to aggravation of atopic dermatitis. We hypothesised that Candida colonisation promotes food allergy, which is known to contribute to a pathogenic response in atopic dermatitis. We tested this using a recently established murine Candida colonisation model. Methods: Candida colonisation in the gastrointestinal tract was established by intragastric inoculation with C albicans in mice fed a synthetic diet. To investigate sensitisation against food antigen, mice were intragastrically administered with ovalbumin every other day for nine weeks, and antiovalbumin antibody titres were measured weekly. To examine gastrointestinal permeation of food antigen, plasma concentrations of ovalbumin were measured following intragastric administration of ovalbumin. Results: Ovalbumin specific IgG and IgE titres were higher in BALB/c mice with Candida colonisation than in normal mice. Gastrointestinal permeation of ovalbumin was enhanced by colonisation in BALB/c mice. Histological examination showed that colonisation promoted infiltration and degranulation of mast cells. Candida colonisation did not enhance ovalbumin permeation in mast cell deficient W/W-v mice but did in congenic littermate control +/+ mice. Reconstitution of mast cells in W/W-v mice by transplantation of bone marrow derived mast cells restored the ability to increase ovalbumin permeation in response to Candida colonisation. Conclusions: These results suggest that gastrointestinal Candida colonisation promotes sensitisation against food antigens, at least partly due to mast cell mediated hyperpermeability in the gastrointestinal mucosa of mice.
• Allergic airway eosinophilia is suppressed in ovalbumin-sensitized brown Norway rats fed raffinose and alpha-linked galactooligosaccharide

  K Sonoyama, H Watanabe, J Watanabe, N Yamaguchi, A Yamashita, H Hashimoto, E Kishino, K Fujita, M Okada, S Mori, S Kitahata, J Kawabata

  JOURNAL OF NUTRITION 135 (3) 538 - 543 0022-3166 2005/03 [Refereed][Not invited]
   

  We recently found that dietary raffinose suppressed allergic airway eosinophilia in ovalbumin-sensitized Brown Norway rats. Using this model in the present study, we compared the efficacy of other oligosaccharides with that of raffinose. Brown Norway rats were immunized s.c. with ovalbumin on d 0 and exposed to aerosolized ovalbumin on d 20; broncho-alveolar lavage fluid was obtained on d 21. In Expt. 1, rats were fed a control diet or diets supplemented with different oligosaccharides (50 g/kg diet, raffinose, a-linked galactooligosaccharide, fructooligosaccharide, and xylooligosaccharide). The number of eosinophils in the fluid was significantly lower in rats fed raffinose and a-linked galactooligosaccharide diets than in those fed the control diet. Dietary fructooligosaccharide and xylooligosaccharide did not affect airway eosinophilia. In Expt. 2, i.p. administration of raffinose and a-linked galactooligosaccharide, but not fructooligosaccharide and xylooligosaccharide, suppressed airway eosinophilia in rats fed the control diet. In Expt. 3, suppression of airway eosinophilia by dietary a-linked galactooligosaccharide occurred in cecectomized rats administered neomycin. Reduced levels of interleukin (IL)-4 and IL-5 mRNA in lung tissue were associated with the suppression of airway eosinophilia. We propose that indigestible oligosaccharides differ in their suppressive effect on allergic airway eosinophilia in ovalbumin-sensitized Brown Norway rats and that the effect appears not to be mediated by intestinal microflora.
• Gastric colonization of Candida albicans differs in mice fed commercial and purified diets

  N Yamaguchi, K Sonoyama, H Kikuchi, T Nagura, T Aritsuka, J Kawabata

  JOURNAL OF NUTRITION 135 (1) 109 - 115 0022-3166 2005/01 [Refereed][Not invited]
   

  It has been difficult to produce persistent colonization by Candida albicans in the gastrointestinal tract of adult mice without the use of antibiotics and immunosuppressants. We hypothesized that diet influences the colonization of C. albicans and tested the hypothesis. BALB/c mice fed either a commercial rodent diet or a nutritionally adequate mixture of purified ingredients were inoculated i.g. with C. albicans (5 x 10(7) cells). Gastrointestinal colonization was examined by fecal culture, tissue culture. and histology. Mice fed the purified diet had a high fecal recovery of C. albicans [5-6log(10) colony forming units (cfu)/g feces] throughout the experimental period (6 wk), and the major site of colonization was the stomach. C. albicans was undetectable in the feces of mice fed the commercial diet 2 wk after inoculation. Immunosuppressants induced systemic dissemination of C. albicans in mice fed the purified diet. The number of lactobacilli and the concentration of organic acids in the stomach were significantly lower in mice fed the purified diet than in those fed the commercial diet. In vitro culture experiments revealed that acetic and lactic acids suppressed the growth of C. albicans. These results suggest drat a reduction in lactobacilli in the stomach of mice fed the purified diet contributed to sustained gastric candidiasis. We therefore propose a novel model of sustained gastric candidiasis by a single i.g. inoculation of C. albicans in healthy adult mice fed a purified diet.
• Oral administration of freeze-dried kefir reduces intestinal permeation of and oral sensitization to ovalbumin in mice

  C Umeda, K Sonoyama, N Yamaguchi, R Saito, K Akashi, H Motoshima, J Kawabata

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 69 (1) 249 - 251 0916-8451 2005/01 [Refereed][Not invited]
   

  An increase in plasma ovalbumin concentrations after intragastric administration of ovalbumin was suppressed by concomitant freeze-dried kefir in BALB/c mice. Serum levels of ovalbumin-specific immunoglobulin G and proliferation of splenic mononuclear cells in mice immunized orally with ovalbumin were suppressed by feeding freeze-dried kefir. We propose that kefir reduces intestinal permeation of food antigen, which contributes to suppression of oral sensitization.
• Reduction of allergic airway eosinophilia by dietary raffinose in Brown Norway rats

  H Watanabe, K Sonoyama, J Watanabe, N Yamaguchi, H Kikuchi, T Nagura, T Aritsuka, K Fukumoto, T Kasai

  BRITISH JOURNAL OF NUTRITION 92 (2) 247 - 255 0007-1145 2004/08 [Refereed][Not invited]
   

  Oral administration of raffinose, a naturally occurring indigestible oligosaccharide, has reportedly ameliorated atopic dermatitis in human subjects although the mechanism is unknown. The present study investigated the effect of dietary raffinose on allergen-induced airway eosinophilia in ovalbumin-sensitised Brown Norway rats as an atopic disease model. Brown Norway rats were immunised by subcutaneous injection with ovalbumin on day 0 and fed either a control diet or the diet supplemented with raffinose (50 g/kg diet). The rats were exposed to aerosolised ovalbumin on day 20, and broncho-alveolar lavage fluid was obtained on the next day. The number of eosinophils in the fluid was significantly lower in the rats fed the raffinose diet than in those fed the control diet. Dietary raffinose significantly reduced IL-4 and IL-5 mRNA levels in lung tissue and tended to lower ovalbumin-specific Ig E levels. Suppression of eosinophilia by dietary raffinose was still observed in caecectomised and neomycin-administered rats, suggesting little contribution by the colonic bacteria to the effect of raffinose. Intraperitoneal administration of raffinose also suppressed eosinophilia. Significant concentrations of raffinose were detected in portal venous and abdominal arterial plasma after the intragastric administration of raffinose. Overall, the findings suggest that dietary raffinose ameliorates allergic airway eosinophilia at least partly via post-absorptive mechanisms in Brown Norway rats.
• The production of hypoallergenic wheat flour and the analysis of its allergy suppressive effects

  J Watanabe, S Tanabe, M Watanabe, H Shinmoto, K Sonoyama

  BIOFACTORS 22 (1-4) 295 - 297 0951-6433 2004 [Refereed][Not invited]
   

  We propose a novel method to produce hypoallergenic wheat flour suitable for patients allergic to wheat by using enzymatic fragmentation with cellulase and actinase. The hypoallergenic flour displayed potent inhibitory activity against allergen absorption and actively suppressed allergic reactions, probably inducing oral tolerance. The results suggest that hypoallergenic wheat flour has allergy-suppressive effects without inducing side effects.
• Isolation of tryptophan as an inhibitor of ovalbumin permeation and analysis of its suppressive effect on oral sensitization

  J Watanabe, K Fukumoto, E Fukushi, K Sonoyama, J Kawabata

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 68 (1) 59 - 65 0916-8451 2004/01 [Refereed][Not invited]
   

  Tryptophan was isolated from rat feces as an active compound against ovalbumin permeation in an in vitro Caco-2 cell model. Tryptophan dose-dependently inhibited ovalbumin permeation with accompanying increase in transepithelial electric resistance, and its inhibitory activity reached a plateau at 10 mm. Brown Norway rats were sensitized by intragastric administration of ovalbumin together with or without tryptophan. Antibody levels specific to ovalbumin in the sera and proliferative responses of spleen mononuclear cells to ovalbumin were significantly lower in rats administered ovalbumin plus tryptophan than those administered ovalbumin alone. These results suggest that tryptophan suppresses oral sensitization to ovalbumin, probably via suppression of ovalbumin absorption from the intestinal tract.
• Effect of enzymatic modification of dietary wheat flour for reducing its allergenicity on oral sensitization to and intestinal absorption of ovalbumin

  R Saito, N Yamaguchi, K Sonoyama, J Kawabata

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 67 (11) 2483 - 2485 0916-8451 2003/11 [Refereed][Not invited]
   

  Increase in plasma immunoglobulin G specific to orally administered ovalbumin in Brown Norway rats was retarded by feeding enzyme-treated wheat flour when compared with untreated flour. Because plasma ovalbumin concentrations after feeding ovalbumin tended to be lower in mice fed enzyme-treated flour than in those fed untreated flour, suppression of ovalbumin absorption may be relevant to retarded sensitization observed in rats.
• Activin A induces phosphorylation of Smad2 but not complex formation of Smad2 with Smad4 in human colon cancer cell line HT-29

  P Pholnukulkit, K Sonoyama, J Kawabata

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 67 (9) 2042 - 2044 0916-8451 2003/09 [Refereed][Not invited]
   

  Western blotting coupled with immunoprecipitation showed that activin A treatment induced phosphorylation of Smad2 but not complex formation of Smad2/4 in human colon cancer-derived HT-29 cells. Because HT-29 cells expressed neither Smad4 mRNA nor Smad4 protein, it is suggested that deletion of Smad4 leads to a defect of formation of Smad2/4 complex upon activin A stimulation in HT-29 cells.
• Upregulation of activin A gene by butyrate in human colon cancer cell lines

  K Sonoyama, P Pholnukulkit, M Toyoda, S Rutatip, T Kasai

  AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY 284 (6) G989 - G995 0193-1857 2003/06 [Refereed][Not invited]
   

  Activin A has been reported to play a role in the progression of colorectal cancer. Because dietary fiber protects against colorectal cancer, we hypothesized that butyrate, a fermentation product of dietary fiber, may affect the expression of activin A in colon cancer cells. Semiquantitative RT-PCR demonstrated that the activin A gene was upregulated by sodium butyrate in the human colon cancer cell lines HT-29 and Caco-2 in a concentration- and time-dependent manner. However, the activin A gene did not respond to sodium butyrate in the human normal colonic cell line FHC, rat normal intestinal epithelial cell (IEC) line IEC-6, and the explant of rat colon. Flow cytometry and agarose gel electrophoresis of genomic DNA revealed that cell cycle arrest and apoptosis were induced by sodium butyrate but not exogenous activin A in HT-29 cells, indicating that activin A could not act as an autocrine factor in colon cancer cells. By assuming that activin A promotes colorectal cancer spread as a paracrine factor, our findings suggest that butyrate could act as a tumor promoter in some circumstances.
• 腸管カンジダ症モデルの作成と解析

  山口 奈津, 園山 慶, 名倉 泰三, 仙波 美博, 川端 潤

  日本栄養・食糧学会大会講演要旨集 (公社)日本栄養・食糧学会 57回 73 - 73 2003/04
• Isolation and structural elucidation of a peptide derived from Edam cheese that inhibits beta-lactoglobulin transport

  S Tanabe, S Tesaki, J Watanabe, E Fukushi, K Sonoyama, J Kawabata

  JOURNAL OF DAIRY SCIENCE 86 (2) 464 - 468 0022-0302 2003/02 [Refereed][Not invited]
   

  A peptide that inhibits beta-lactoglobulin absorption in an in vitro Caco-2 cell model was isolated from Edam cheese. By H-1-NMR and N-terminal amino acid analyses, the active compound was identified as Asp-Lys-Ile-His-Pro-Phe. The sequence of the hexapeptide is the same as the amino acid sequence of residues 47-52 of beta-casein. The hexapeptide shows remarkable inhibition of beta-lactoglobulin transport at a concentration of 10(-7) M. The possibility exists that this peptide can be applied practically to the prevention of milk-sensitive allergy.
• An active compound against allergen absorption in hypoallergenic wheat flour produced by enzymatic modification

  S Tesaki, J Watanabe, S Tanabe, K Sonoyama, E Fukushi, J Kawabata, M Watanabe

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 66 (9) 1930 - 1935 0916-8451 2002/09 [Refereed][Not invited]
   

  Hypoallergenic wheat flour produced by modification with cellulase and actinase showed inhibitory activity against ovalbumin permeation in an in vitro model by using the Caco-2 cell monolayer. The activity was found in the cellulase preparation used for producing the flour. An active compound was isolated by HPLC and identified as Trp-Ser-Asn-Ser-Gly-Asn-Phe-Val-Gly-Gly-Lys by H-1-NMR data and Edman degradation. The undecapeptide, some oligopeptides with the N-terminal sequences and Trp ethyl ester showed activity at 10(-7) m, acetyl Trp being active at 10(-2) M. These data suggest that the Trp residue without a free carboxyl group would be required for the inhibitory activity of ovalbumin absorption through the intestinal tract.
• ラフィノースが肥満細胞機能に及ぼす影響

  山口 奈津, 葛西 隆則, 園山 慶

  日本栄養・食糧学会大会講演要旨集 (公社)日本栄養・食糧学会 56回 119 - 119 2002/06
• IgE-reactive 60 kDa glycoprotein occurring in wheat flour

  J Watanabe, S Tanabe, K Sonoyama, M Kuroda, M Watanabe

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 65 (9) 2102 - 2105 0916-8451 2001/09 [Refereed][Not invited]
   

  A new IgE-reactive glycoprotein with a molecular size of 60 kDa was isolated from wheat flour. The N-terminal amino acid sequence of the protein was LDPDESEXVTRYFRIR. The 8th amino acid residue would have been Asn to which the peroxidase-type glycochain was attached. The IgE-binding activity of the glycoprotein was rendered negligible by the enzymatic treatment applied for hypoallergenic flour production.
• Cholesterol-lowering effects of maitake (Grifola frondosa) fiber, shiitake (Lentinus edodes) fiber, and enokitake (Flammulina velutipes) fiber in rats

  M Fukushima, T Ohashi, Y Fujiwara, K Sonoyama, M Nakano

  EXPERIMENTAL BIOLOGY AND MEDICINE 226 (8) 758 - 765 0037-9727 2001/09 [Refereed][Not invited]
   

  The effects of mushroom fibers on serum cholesterol and hepatic low-density lipoprotein (LDL) receptor mRNA in rats were investigated. Rats were fed a cholesterol-free diet with 50 g/kg cellulose powder (CP), 50 g/kg maitake (Grifola frondosa) fiber (MAF), 50 g/kg shiitake (Lentinus edodes) fiber (SF), or 50 g/kg enokitake (Flammulina velutipes) fiber (EF) for 4 weeks. There were no significant differences in the body weight, food intake, liver weight, cecum weight, and cecum pH among the groups. Cecal acetic acid, butyric acid, and total short-chain fatty acid (SCFA) concentrations in the SF and EF groups were significantly higher than those in the other groups. The serum total cholesterol concentration in the CP group was significantly higher than that in the MAF and EF groups. The very LDL (VLDL) + intermediate-density lipoprotein (IDL) + LDL-cholesterol concentration in the CP group was significantly higher than that in the MAF, SF, and EF groups, whereas the high-density lipoprotein (HDL)-cholesterol concentration in the EF group was significantly lower than that in the other groups at the end of the 4-week feeding period. The hepatic LDL receptor mRNA level in the EF group was significantly higher than that in the CP group. The fecal cholesterol excretion in the MAF, SF, and EF groups was significantly higher than that in the CP group. The results of this study demonstrate that MAF and EF lowered the serum total cholesterol level by enhancement of fecal cholesterol excretion, and in particular, by enhancement of hepatic LDL receptor mRNA in EF group.
• Consumption of hypoallergenic flour prevents gluten-induced airway inflammation in brown Norway rats

  J Watanabe, S Tanabe, M Watanabe, T Kasai, K Sonoyama

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 65 (8) 1729 - 1735 0916-8451 2001/08 [Refereed][Not invited]
   

  Brown Norway rats were immunized with gluten, and then fed a diet containing hypoallergenic flour or an amino acid mixture. The rats were then made to inhale a solubilized gluten to induce gluten-specific bronchial asthma. The antibody levels in the serum of rats were measured by ELISA, and cell counts were done on cytospin preparations of bronchoalveolar lavage fluid. Body weight was decreased after allergen challenge in rats fed the amino acid mixture but not in rats fed the hypoallergenic flour. Antibody levels in the serum were significantly lower in rats fed hypoallergenic flour than in those fed the amino acid mixture. Differential cell counts in the bronchoalveolar lavage fluid showed that the numbers of eosinophils, lymphocytes, and neutrophils were significantly lower in rats fed the hypoallergenic flour than in those fed the amino acid mixture. These results suggest that hypoallergenic flour actively suppresses the allergic reactions; probably by inducing oral tolerance.
• Investigation of gene expressions related to cholesterol metabolism in rats fed diets enriched in n-6 or n-3 fatty acid with a cholesterol after long-term feeding using quantitative-competitive RT-PCR analysis

  M Fukushima, K Shimada, T Ohashi, H Saitoh, K Sonoyama, M Sekikawa, M Nakano

  JOURNAL OF NUTRITIONAL SCIENCE AND VITAMINOLOGY 47 (3) 228 - 235 0301-4800 2001/06 [Refereed][Not invited]
   

  We have developed a method to quantitate hepatic apolipoprotein (apo) B, LDL receptor, 3-hyddroxy-3-methylglutary coenzyme A reductase (HMG-CoA reductase) and cholesterol 7 alpha -hydroxylase mRNA expression in rats fed a cholesterol-enriched diet after long-term feeding using competitive RT-RCR. Rats (8 wk of age) fed a conventional diet were shifted to diets containing 10%, perilla oil (PEO, oleic acid + linoleic acid + alpha -linolenic acid), borage oil (BRO, oleic acid + linoleic acid + gamma -linolenic acid), evening primrose oil (EPO, linoleic acid + gamma -linolenic acid), mixed oil (MIO, oleic acid+linoleic acid+ gamma -linolenic acid+ alpha -linolenic acid), or palm oil (PLO, palmitic acid +oleic acid+ linoleic acid) with 0.5% cholesterol for 15 wk. There were no significant differences in the food intake and body weight gain among the groups. The liver weight in the PEO and PLO groups was significantly higher than other groups. The serum total cholesterol and very low density lipoprotein (VLDL) + intermediate density lipoprotein (IDL)+ low density lipoprotein (LDL)-cholesterol concentrations were consistently higher in PLO group than in the other groups. The serum high density lipoprotein cholesterol concentration was significantly lower in the PEO group than in the other groups. The liver cholesterol concentration group was significantly higher in the PEO than in the other groups. There were no significant differences in the hepatic LDL receptor mRNA level among the groups. Hepatic apo B. HMG-CoA reductase and cholesterol 7 alpha -hydroxylase mRNA levels were not affected by the experimental conditions. However, hepatic cholesterol 7 alpha -hydroxylase mRNA level in the PEO and MIO groups tended to be higher than in the other groups. The fecal cholesterol extraction was significantly higher in the MIO and PLO groups than in the PEO and EPO groups and the total bile acid extraction was significantly higher in the PEO and MIO groups than in the PLO group. The results of this study demonstrated that both n-6 fatty acid and n-3 fatty acids such as gamma -linolenic acid and alpha -linolenic acid lowered serum total cholesterol and VLDL+IDL+LDL-cholesterol concentrations of rats in the presence of excess cholesterol in the diet compared with dietary saturated fatty acid.
• 低アレルゲン化小麦粉によるアレルゲンの腸管透過抑制効果

  渡辺道子, 渡辺純, 田辺創一, 福士江里, 園山慶, 川端潤

  日本農芸化学会誌 75 283  0002-1407 2001/03/05 [Refereed][Not invited]
  
• エダムチーズに含まれるアレルゲン腸管透過抑制ペプチド

  田辺創一, 渡辺純, 福士江里, 園山慶, 川端潤, 渡辺道子

  日本農芸化学会誌 75 283  0002-1407 2001/03/05 [Refereed][Not invited]
  
• Effects of diets enriched in n-6 or n-3 fatty acids on cholesterol metabolism in older rats chronically fed a cholesterol-enriched diet

  M Fukushima, T Ohhashi, S Ohno, H Saitoh, K Sonoyama, K Shimada, M Sekikawa, M Nakano

  LIPIDS 36 (3) 261 - 266 0024-4201 2001/03 [Refereed][Not invited]
   

  Hypocholesterolemic effects in older animals after long-term feeding are unknown. Therefore, aged rats (24 wk Of age) fed a conventional diet were shifted to diets containing 10% perilla oil [PEO; oleic acid + linoleic acid + alpha -linolenic acid; n-6/n-3, 0.3; polyunsaturated fatty acid/saturated fatty acid (P/S), 9.6], borage oil [oleic acid + linoleic acid + alpha -linolenic acid; n-6/n-3, 15.1; P/S, 5.3], evening primrose oil (EPO; linoleic acid + gamma -linolenic acid; P/S, 10.5), mixed oil (MIO; oleic acid + linoleic acid + gamma -linolenic acid + alpha -linolenic acid; n-6/n-3, 1.7; P/S, 6.7), or palm oil (PLO; palmitic acid + oleic acid + linoleic acid; n-6/n-3, 25.3; P/S, 0.2) with 0.5% cholesterol for 15 wk in this experiment. There were no significant differences in the food intake and body weight gain among the groups. The liver weight in the PEO (n-6/n-3, 0.3) group was significantly higher than those of other groups in aged rats. The serum total cholesterol and very low density lipoprotein (VLDL) + intermediate density lipoprotein (IDL) + low density lipoprotein (LDL)-cholesterol concentrations of the PLO (25.3) group were consistently higher than those in the other groups. The serum high density lipoprotein cholesterol concentrations of the PEO (0.3) and EPO groups were significantly lower than in the other groups at the end of the 15-wk feeding period. The liver cholesterol concentration of the PLO (25.3) group was significantly higher than those of other groups. There were no significant differences in the hepatic LDL receptor mRNA level among the groups. Hepatic apolipoprotein (apo) B mRNA levels were not affected by the experimental conditions. The fecal neutral steroid excretion of the PLO (25.3) group tended to be low compared to the ether groups. The results of this study demonstrate that both n-6 fatty acid and n-3 fatty acids such as gamma -linolenic acid and cx-linolenic acid inhibit the increase of serum total cholesterol and VLDL + IDL + LDL-cholesterol concentrations of aged rats in the presence of excess cholesterol in the diet compared with dietary saturated fatty acid.
• Low density lipoprotein receptor mRNA in rat liver is affected by resistant starch of beans

  M Fukushima, T Ohashi, M Kojima, K Ohba, H Shimizu, K Sonoyama, M Nakano

  LIPIDS 36 (2) 129 - 134 0024-4201 2001/02 [Refereed][Not invited]
   

  The effects of resistant starches of beans on serum cholesterol and hepatic low density lipoprotein (LDL) receptor mRNA in rats were investigated. Rats were fed a cholesterol-free diet with 150 g/kg corn starch (CS), 150 g/kg adzuki (Vigna angularis) starch (AS), 150 g/kg kintoki (Phaseolus vulgaris, variety) starch (KS), or 150 g/kg tebou (P. vulgaris, variety) starch (TS) for 4 wk. There were no significant differences in body weight among groups through the experimental period. The liver weight in the CS group was 1.1-1.2 times higher than that in the AS, KS, and TS groups. The cecum weight in the TS was 1.4 times higher than that in the CS group, and the cecal pH in the CS group was significantly higher than in the other groups. The serum total cholesterol, very low density lipoprotein + intermediate density lipoprotein + LDL-cholesterol and high density lipoprotein (HDL)-cholesterol concentrations in the bean starch groups were significantly lower than those in the CS group through the feeding period. The total cholesterol/HDL-cholesterol ratio in the bean starch groups was also significantly lower than that in the CS group at the end of the 4-wk feeding period. The hepatic cholesterol concentration in the TS group was significantly higher than in the CS group at the end of the 4-wk feeding period. The relative quantity of hepatic apo B mRNA in the AS group was 1.2 times higher than that in the CS group, and the hepatic LDL receptor mRNA levels in the AS and TS groups were 1.8-2.0 times higher than that in the CS group. The results of this study demonstrate that AS, KS, and TS lowered the serum total cholesterol level by enhancing the hepatic LDL receptor mRNA level.
• Novel method for producing hypoallergenic wheat flour by enzymatic fragmentation of the constituent allergens and its application to food processing

  M Watanabe, J Watanabe, K Sonoyama, S Tanabe

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 64 (12) 2663 - 2667 0916-8451 2000/12 [Refereed][Not invited]
   

  A novel method is proposed to produce hypoallergenic wheat flour suitable for patients allergic to wheat. Wheat flour was mixed with a cellulase solution, and the mixture was incubated at 50 degreesC for 1 h to hydrolyze the carbohydrate allergens. The hydrolysate was further incubated with actinase at 40 degreesC for 1 h while gently stirring to decompose the proteinaceous allergens. The product was evaluated for its allergenicity by an enzyme-linked immunosorbent assay, the results of which suggested negative allergenicity in most cases. The product changed to a batter state that was difficult to process by the usual methods. Gelatinization of the starch in the product and the addition of a surfactant were beneficial for food processing.
• Hepatic LDL receptor mRNA in rats is increased by dietary mushroom (Agaricus bisporus) fiber and sugar beet fiber

  M Fukushima, M Nakano, Y Morii, T Ohashi, Y Fujiwara, K Sonoyama

  JOURNAL OF NUTRITION 130 (9) 2151 - 2156 0022-3166 2000/09 [Refereed][Not invited]
   

  Plasma cholesterol concentration is reduced by feeding some dietary fibers and mushroom fruit body, but the mechanism is not fully understood. We examined the effects of mushroom (Agaricus bisporus) fiber and sugar beet fiber on serum cholesterol and hepatic LDL receptor mRNA in rats. Rats were fed a cholesterol-free diet with 50 g/kg cellulose powder (CP), 50 g/kg mushroom (Agaricus bisporus) fiber (MSF) or 50 g/kg sugar beet fiber (BF) for 4 wk. There were no significant differences in the body weight, food intake and cecum weight among the groups. The relative liver weight in the CP group was significantly greater than that in the MSF and BF groups. The cecal pH in the CP and MSF groups was significantly higher than that in the BF group. Cecal acetic acid, butyric acid and total short-chain fatty acid (SCFA) concentrations in the BF group were significantly higher than those in the other groups. The serum total cholesterol, VLDL + intermediate density lipoprotein (IDL) + IDL cholesterol concentrations in the CP group were significantly greater than those in the MSF and BF groups. The HDL cholesterol concentration in the MSF group was significantly lower than that in the CP group. The hepatic LDL receptor mRNA level in the MSF and BF groups was significantly higher than that in the CP group. The results of this study demonstrate that mushroom fiber and sugar beet fiber lowered the serum total cholesterol level by enhancement of the hepatic LDL receptor mRNA.
• Non-effect of hexamethonium, a ganglionic blocker, on the response of ileal apolipoprotein A-IV mRNA following a massive small bowel resection in rats

  K Sonoyama, R Fujiwara, T Kasai

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 64 (6) 1282 - 1284 0916-8451 2000/06 [Refereed][Not invited]
   

  An intravenous infusion of hexamethonium, a ganglionic blocker, did not affect the increase in the apolipoprotein A-IV mRNA level in the residual ileum following a massive small bowel resection in unrestrained conscious rats. The result suggests that upregulation of the apolipoprotein A-IV gene in the residual ileum is not mediated by a neural pathway, including the nicotinic synapse route.
• Intravenous infusion of hexamethonium and atropine but not propranolol diminishes apolipoprotein A-IV gene expression in rat ileum

  K Sonoyama, K Tajima, R Fujiwara, T Kasai

  JOURNAL OF NUTRITION 130 (3) 637 - 641 0022-3166 2000/03 [Refereed][Not invited]
   

  To clarify the role of neural factors in the regulation of apolipoprotein (apo) A-IV expression in the small intestine, we investigated the effect of neural blockers on mRNA levels of apo A-IV in rat small intestine. Either ganglionic blocker (hexamethonium), cholinergic blocker (atropine) or beta-adrenergic blocker (propranolol) was infused intravenously to unrestrained conscious rats for 8 h, and then total RNA was isolated from the small intestine and analyzed using Northern hybridization. Apo A-IV mRNA levels in the ileum were significantly lower in hexamethonium- or atropine-infused rats than in saline- (control) or propranolol-infused rats. Immunoblot analysis showed no difference in plasma apo A-IV concentrations between hexamethonium- and saline-infused groups. The lower mRNA levels of apo A-IV in the ileum of hexamethonium-infused rats were observed even in bile-drained rats, indicating that the lower expression was not due to any changes in bile availability. The ileal apo A-IV mRNA levels were significantly higher in rats infused with lipid emulsion into the ileum than in rats infused with glucose-saline, and the concomitant infusion of intravenous hexamethonium did not affect the higher levels of apo A-IV mRNA. These results suggest that the basal expression of the ileal A-IV gene is at least partially regulated in a site-specific manner by cholinergic neurons.
• Peptide YY stimulates the expression of apolipoprotein A-IV gene in Caco-2 intestinal cells

  K Sonoyama, K Suzuki, T Kasai

  PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE 223 (3) 270 - 275 0037-9727 2000/03 [Refereed][Not invited]
   

  The effect of peptide YY, a gastrointestinal hormone, on the expression of the apolipoprotein A-IV gene in the intestinal epithelial cell line Caco-2 was examined by semiquantitative RT-PCR followed by Southern hybridization with an inner oligonucleotide probe. Apolipoprotein A-IV mRNA levels were increased in response to peptide YY in a dose- and time-dependent fashion, Western blotting revealed that the exogenous peptide YY increased the intracellular concentration of apolipoprotein A-IV, in contrast, apolipoprotein A-I, B, and C-III mRNA did not respond to peptide YY. Differentiated Caco-2 cells expressed Y1- but not Y2- and YS-receptor subtype mRNA, The present results suggest that peptide YY modulates apolipoprotein A-IV gene expression, likely via the Y1-receptor subtype in intestinal epithelial cells.
• Gene expression of activin, activin receptors, and follistatin in intestinal epithelial cells

  K Sonoyama, S Rutatip, T Kasai

  AMERICAN JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER PHYSIOLOGY 278 (1) G89 - G97 0193-1857 2000/01 [Refereed][Not invited]
   

  Gene expression of activin, activin receptors, and follistatin was investigated in vivo and in vitro using semiquantitative RT-PCR in intestinal epithelial cells. Rat jejunum and the intestinal epithelial cell line IEC-6 expressed mRNA encoding the beta A-subunit of activin, alpha-subunit of inhibin, activin receptors IB and IIA, and follistatin. An epithelial cell isolation study focused along the crypt-villus axis in rat jejunum showed that beta A mRNA levels were eight- to tenfold higher in villus cells than in crypt cells. Immunohistochemistry revealed the expression of activin A in upper villus cells. The human intestinal cell line Caco-2 was used as a differentiation model of enterocytes. Four- to fivefold induction of beta A mRNA was observed in postconfluent Caco-2 cells grown on filter but not in those cells grown on plastic. In contrast, follistatin mRNA was seen to be reduced after reaching confluence. Exogenous activin A dose-dependently suppressed the proliferation and stimulated the expression of apolipoprotein A-TV gene, a differentiation marker, in IEC-B cells. These results suggest that the activin system is involved in the regulation of such cellular functions as proliferation and differentiation in intestinal epithelial cells.
• Apolipoprotein B mRNA editing in the liver and small intestine of rats fed on beet fiber, soy protein, and fish oil

  R Miyakawa, K Sonoyama, T Kasai

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 63 (7) 1315 - 1317 0916-8451 1999/07 [Refereed][Not invited]
   

  Apolipoprotein B mRNA editing was investigated in the liver and small intestine of rats fed on beet fiber, soy protein, or fish oil as plasma cholesterol-reducing agents. The diets had no influence on the editing in both the liver and intestine, despite their cholesterol-lowering action. The results suggest that apo B mRNA editing is not involved in the cholesterol-lowering effect of these diets.
• Different responses of apolipoprotein A-I, A-IV, and B gene expression during intestinal adaptation to a massive small bowel resection in rats

  K Sonoyama, Y Aoyama

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 61 (11) 1810 - 1813 0916-8451 1997/11 [Refereed][Not invited]
   

  Gene expression of apolipoproteins (ape) A-I, A-IV, and B, the predominant protein components of chylomicrons, was investigated in the residual ileum after a massive small bowel resection in rats. A Northern blot analysis showed that the ape A-IV mRNA level, but not the ape A-I and B mRNA levels, in the ileum was significantly higher in the resected rats than in the sham-operated rats 24h and 2 wk post-surgery. RT-PCR coupled with a primer extension assay revealed that the ape B-48 mRNA/apo B-100 mRNA ratio, i.e., ape B mRNA editing, in the ielum was unchanged by the resection. It is thus concluded that, among the major intestinal apolipoproteins, ape A-IV is the only one whose gene expression is influenced by loss of the proximal intestine.
• Changes in the number and apoptosis of epithelial cells in the colorectum of wheat bran-fed rats soon after administering 1,2-dimethylhydrazine

  S Ishizuka, K Sonoyama, T Kasai

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 61 (8) 1337 - 1341 0916-8451 1997/08 [Refereed][Not invited]
   

  We investigated the effect of dietary wheat bran (Wb) on colonic tumorigenesis soon after a single administration of 1,2-dimethylhydrazine (DMH). Rats that had been fed on either a fiber-free diet or a 20% Wb diet were injected with 1,2-dimethylhydrazine (20 mg/kg body weight). At 6 h, 12 h, 1 d, 3 d, or 7 d after the injection, the colorectum was excised for histological analyses, The number of crypt cells more rapidly recovered in the 20% Wb group than in the fiber-free group after its temporary reduction by injection of DMH. At 6h after the DMH treatment, the apoptotic cells were significantly greater in number in the fiber-free group than in the 20% Wb group, In contrast, those in distal colon were significantly fewer in the fiber-free group than in the 20% Wb group at 7 d after the treatment, These results suggest that the ingestion of Wb affected the turnover of colonic epithelial cells and would thereby bring about a protective effect against DMH-induced tumorigenesis.
• Stimulating effect of ileal pancreaticobiliary secretion on ileal apolipoprotein A-IV mRNA expression in fasted rats

  K Sonoyama, R Fujiwara, Y Aoyama

  BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY 61 (5) 887 - 889 0916-8451 1997/05 [Refereed][Not invited]
   

  The effect of pancreaticobiliary secretion on the intestinal expression of the apo A-IV gene was examined in fasted rats. Pancreaticobiliary diversion, but not biliary diversion alone, into the ileum increased the ileal apo A-IV mRNA expression by 24 h post-operation. Jejunal apo A-IV mRNA was reduced by biliary exclusion. The data suggest that the biliary constituent plays an important role in the apo A-IV gene expression in the entire length of the small intestine, and that up-regulation of the apo A-IV gene requires exocrine pancreatic in addition to biliary secretion.
• 小麦ふすま摂取ラット大腸における1,2-dimethylhydrazine投与後の細胞死と増殖の経時的変化 : 動物

  石塚 敏, 園山 慶, 葛西 隆則

  日本農藝化學會誌 社団法人日本農芸化学会 70 (0) 30 - 30 0002-1407 1996/03/05 [Refereed][Not invited]
  
• ラット小腸部分切除モデルを用いた小腸apo B mRNA editing protein遺伝子発現の解析 : 動物

  園山 慶, 仁木 良哉

  日本農藝化學會誌 社団法人日本農芸化学会 70 (0) 32 - 32 0002-1407 1996/03/05 [Refereed][Not invited]
  
• Upregulation of apolipoprotein A-IV gene expression in residual ileum after massive small bowel resection requires the biliary secretion in rats

  K Sonoyama, Y Nakamura, S Kiriyama, R Niki

  PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE 211 (3) 273 - 280 0037-9727 1996/03 [Refereed][Not invited]
   

  Small bower resection results in adaptation of residual intestine, but little is known about the postresectional response of apolipoproteins synthesized mainly in the small intestine, We have investigated the postresectional response of apolipoprotein A-I and A-IV gene expression in residual ileum and assessed the mechanism of response, particularly the role of biliary secretion, Time course of changes in apolipoprotein A-I and A-IV mRNA levels was examined by Northern blotting in the residual terminal ileum for 24 hr after 85% jejunoileal resection in fasted rats, Localization of these mRNAs was studied using in site hybridization histochemistry. Effect of biliary diversion on the postresectional response of mRNAs and proteins was estimated by Northern blotting and immunoblotting, respectively, Apolipoprotein A-IV mRNA began increasing at 1 hr postresection, achieved a maximum by 12 hr (5-fold increase) and remained stable to 24 hr, while apolipoprotein A-I mRNA did not change, Apolipoprotein A-IV mRNA accumulated predominantly in the upper part of ileal villi and increased its intensity postresection, and apolipoprotein A-I mRNA was detected in the villus base to tip. The postresectional increase in apolipoprotein A-IV mRNA and protein was completely abolished by concurrent biliary diversion, The results suggest that the enterocytes in the ileal villi rapidly adapt to massive small bowel resection by increase in apolipoprotein A-IV gene expression which is mediated by biliary constituents but not luminal nutrients.
• ラット小腸広範囲切除後の残存回腸アポリポ蛋白A-I及びA-IV遺伝子発現の組織学的解析 : 動物

  園山 慶, 中村 吉孝, 桐山 修八, 仁木 良哉

  日本農藝化學會誌 社団法人日本農芸化学会 69 (0) 345 - 345 0002-1407 1995/07/05 [Refereed][Not invited]
  
• EXPRESSION OF APOLIPOPROTEIN-A-I MESSENGER-RNA IN LIVER AND INTESTINE OF CECECTOMIZED RATS FED BEET FIBER

  K SONOYAMA, S KIRIYAMA, R NIKI

  JOURNAL OF NUTRITIONAL BIOCHEMISTRY 6 (7) 380 - 384 0955-2863 1995/07 [Refereed][Not invited]
   

  Sham-operated and cecectomized rats were fed a cholesterol-free diet with no added fiber (fiber-free) for 10 days, followed by the diet of 15% beet fiber for 10 days. The plasma cholesterol concentrations in rats fed the fiber-free diet were not significantly different between sham-operated and cecectomized groups. Plasma cholesterol concentrations in sham-operated rats were decreased by feeding the beet fiber diet, whereas those in cecectomized rats did not change. Final plasma total cholesterol concentrations in cecectomized rats were significantly higher than in sham-operated rats. This difference was due mainly to lower HDL cholesterol concentrations. The cecectomized rats also tended to have higher apolipoprotein A-I concentrations in plasma. Northern blot analysis revealed that the relative concentrations of ileal apolipoprotein A-I mRNA were the same in the two groups, while hepatic apolipoprotein A-I mRNA levels were significantly higher in cecectomized rats than in sham-operated rats. These data demonstrate that the cecectomy abolished the hepatic apolipoprotein A-I mRNA-lowering effect of dietary beet fiber, and it is suggested that the cecum plays an important role in the regulation of hepatic apolipoprotein A-I expression which seems to be responsible for the hypocholesterolemic effect of dietary beet fiber.
• ADAPTIVE RESPONSE OF APOLIPOPROTEIN-A-I AND A-IV MESSENGER-RNA IN RESIDUAL ILEUM AFTER MASSIVE SMALL-BOWEL RESECTION IN RATS

  K SONOYAMA, S KIRIYAMA, R NIKI

  JOURNAL OF NUTRITIONAL SCIENCE AND VITAMINOLOGY 41 (2) 253 - 264 0301-4800 1995/04 [Refereed][Not invited]
   

  The response of apolipoproteins, which are synthesized mainly in the small intestine, after small bowel resection has not been documented. In this study, we investigated the effect of small bowel resection on the expression of apolipoprotein mRNA in the residual ileum in rats. Wistar rats underwent either an 85% jejunoileal resection or a sham-operation. Plasma concentrations of total and HDL-cholesterol, apolipoprotein A-I and A-IV were measured on days 0, 3, 6, 9, 12, and 16 (surgery on day 1). The abundances of apolipoprotein mRNA in the residual ileum and liver on day 16 were determined. Plasma levels of total and HDL-cholesterol and apolipoprotein A-I and A-IV in resected rats were significantly lower than in sham-operated rats on days 3 and 6. Resected rats showed a significant increase in ileal apolipoprotein A-I (1.2-fold) and A-IV (3.2-fold) mRNA compared with sham-operated animals. Hepatic apolipoprotein mRNA were the same between two groups. These data suggest that the residual ileum adapts to jejunoileal resection by selective increases in apolipoprotein A-I and A-IV expression at a pretranslational stage. The recoveries of apolipoprotein A-I and A-IV in plasma appear to depend, at least in part, on the increased expression of these apolipoproteins in the residual ileum.
• APOLIPOPROTEIN MESSENGER-RNA IN LIVER AND INTESTINE OF RATS IS AFFECTED BY DIETARY BEET FIBER OR CHOLESTYRAMINE

  K SONOYAMA, H NISHIKAWA, S KIRIYAMA, R NIKI

  JOURNAL OF NUTRITION 125 (1) 13 - 19 0022-3166 1995/01 [Refereed][Not invited]
   

  Rats were fed a cholesterol-free diet with no added fiber (fiber-free) or with 15 g/100 g beet fiber or 5 g/100 g cholestyramine for 14 d. Final plasma total cholesterol concentrations were significantly lower in rats fed beet fiber than in those fed fiber-free or cholestyramine diets. This difference was due mainly to lower HDL cholesterol concentrations. The group fed beet fiber also tended (P < 0.1) to have lower apolipoprotein A-I concentration in plasma. Northern blot analysis revealed that the relative concentrations of jejunal apolipoprotein A-I and A-IV mRNA were the same in all groups, whereas ileal apolipoprotein A-I and A-IV mRNA levels were significantly lower in rats fed beet fiber or cholestyramine than in those fed the fiber-free diet. Hepatic apolipoprotein E mRNA concentrations were the same in all groups, but apolipoprotein A-I mRNA levels were significantly lower in rats fed beet fiber than in those fed the other diets. Apolipoprotein AIV mRNA tended (P < 0.1) to be lower in rats fed the beet fiber diet. These data suggest that the hypocholesterolemic effect of dietary beet fiber is associated with diminished expression of the hepatic apolipoprotein A-I gene.
• FOOD-DEPRIVATION INCREASES APOPTOTIC CELL COUNTS INDUCED BY 1,2-DIMETHYLHYDRAZINE IN RAT DESCENDING COLONIC AND RECTAL CRYPTS

  S ISHIZUKA, K SONOYAMA, R NIKI

  JOURNAL OF NUTRITIONAL SCIENCE AND VITAMINOLOGY 40 (4) 363 - 369 0301-4800 1994/08 [Refereed][Not invited]
   

  The frequency of apoptosis after treatment with 1,2-dimethylhydrazine (DMH) was counted in the descending colonic and rectal crypts of food-deprived and fed rats. Food-deprived or fed rats were subcutaneously injected with DMH (100 mg/kg body weight). Six hours after the injection, apoptotic cells were observed in crypt regions by light microscopy. The incidence of DMH-induced apoptosis in food-deprived rats was significantly higher than in fed rats. The incidence appeared to be higher in descending colon than in rectum. PAS staining revealed that DMH treatment lowered mucin secretion in crypts, which was substantially lowered by food deprivation. The effect of food deprivation on apoptosis induced by DMH may be due to the decrease in mucus barrier against DMH.
• BILE DIVERSION LOWERS APOLIPOPROTEIN-A-I AND A-IV MESSENGER-RNA LEVELS IN RAT ILEUM

  K SONOYAMA, H NISHIKAWA, S KIRIYAMA, R NIKI

  JOURNAL OF NUTRITIONAL SCIENCE AND VITAMINOLOGY 40 (4) 343 - 352 0301-4800 1994/08 [Refereed][Not invited]
   

  We recently reported that cholestyramine (a bile acid sequestrant) lowered ileal apolipoprotein A-I mRNA level in rats. To obtain further information about this phenomenon, in this study, we investigated whether bile diversion lowers apolipoprotein A-I mRNA level in the ileum of rats. Bile-diverted rats were fed a diet with no added Na taurocholate (control diet) or with 0.4% Na taurocholate for 7 days. Sham-operated rats were also fed the control diet for the same period. Northern blot analysis revealed that the relative concentrations of jejunal apolipoprotein A-I and A-IV mRNA at the end of experimental period did not differ between groups while ileal apolipoprotein A-I and A-IV mRNA concentrations were significantly lower in bile-diverted rats fed the control and Na taurocholate-containing diets than in sham-operated rats. Plasma total and HDL cholesterol concentrations were the same in all groups. Relative concentration of apolipoprotein A-I in plasma also did not change. These results suggest that the bile plays an important role in ileal apolipoprotein gene expression at the pretranslational stage, but it is still unclear whether the effector is the bile acid or not. The unchanged concentration of plasma apolipoprotein A-I may have resulted from the constant secretion independent of synthesis in the intestine or the larger contribution from the liver which is another principal site for apolipoprotein A-I expression.
• EFFECT OF DIETARY-PROTEIN LEVEL ON INTESTINAL AMINOPEPTIDASE ACTIVITY AND MESSENGER-RNA LEVEL IN RATS

  K SONOYAMA, S KIRIYAMA, R NIKI

  JOURNAL OF NUTRITIONAL BIOCHEMISTRY 5 (6) 291 - 297 0955-2863 1994/06 [Refereed][Not invited]
   

  To investigate the mechanism of adaptive response of intestinal aminopeptidase on the dietary protein level, we determined the aminopeptidase activity and mRNA level in the intestinal mucosa of rats with different dietary protein levels. In the first experiment, the specific activity of aminopeptidase in the homogenate of intestinal mucosa from rats fed a 60% casein diet for 7 days was significantly higher than that from rats fed a 20% casein diet. A larger difference between the two groups was observed in the ileum than in the jejunum. In contrast to aminopeptidase, the specific activity of sucrase in the ileum of rats fed a 60% casein diet was significantly lower than that of rats fed a 20% casein diet. The activities of sucrase in the jejunum and alkaline phosphatase in the jejunum and ileum did not change between the two groups. Northern blot analysis of intestinal RNA using 1.1 kb aminopeptidase N cDNA as a probe did not show a significant difference in the level of aminopeptidase N mRNA between two dietary groups. In the next experiment the aminopeptidase activity and mRNA level in the ileal mucosa were determined in rats fed 0, 10, 20, 40, and 60% casein diets for 7 days. While the specific activity of aminopeptidase increased in response to the dietary protein level, sucrase and alkaline phosphatase activities did not change. However we did not observe any change in the level of ileal aminopeptidase N mRNA in all groups, and there was no significant correlation between the specific activity and the mRNA abundance. These data indicate that the catalytic activity of intestinal aminopeptidase responds to the dietary protein level, but the level of mRNA is preserved even under the protein malnourished condition. The data also suggest that translational and/or posttranslational regulations for the expression of intestinal aminopeptidase are necessary for the rapid response to changes in intraluminal nutrients.
• CHOLESTYRAMINE AND A FAT-FREE DIET LOWER APOLIPOPROTEIN A-IV MESSENGER-RNA IN JEJUNUM AND CHOLESTYRAMINE LOWERS APOLIPOPROTEIN-A-I MESSENGER-RNA IN ILEUM OF RATS

  K SONOYAMA, H NISHIKAWA, S KIRIYAMA, R NIKI

  JOURNAL OF NUTRITION 124 (5) 621 - 627 0022-3166 1994/05 [Refereed][Not invited]
   

  To investigate the effect of bile acids or dietary lipid on the expression of intestinal apolipoproteins, the mRNA levels of apolipoprotein A-I and A-IV in the intestine from rats fed a diet containing cholestyramine or a fat-free diet were compared with those from rats fed a control diet containing 25% casein and 5% corn oil. Plasma total cholesterol concentration was lower after 16 h in rats fed a diet containing cholestyramine or a fat-free diet than in rats fed a control diet. In rats fed the fat-free diet, HDL cholesterol concentration also was lower than in those fed the control diet. The pool of bile acid in intestinal contents was significantly lower in rats fed cholestyramine than in both other groups. The relative abundance of jejunal apolipoprotein A-I mRNA did not differ between groups. Jejunal apolipoprotein A-IV mRNA abundance was significantly lower than in controls in rats fed the fat-free and cholestyramine-containing diets. Abundance of apolipoprotein A-I mRNA in ileal mucosa was comparable to controls in rats fed a fat-free diet but was significantly lower in rats fed cholestyramine. Ileal apolipoprotein A-IV mRNA tended to be lower in rats fed cholestyramine and a fat-free diet than in controls. We propose that decreased absorption of dietary lipid may modulate changes in jejunal apolipoprotein A-IV mRNA level and low levels of bile acids in the lumen may modulate changes in ileal apolipoprotein A-I mRNA level.
• CHOLESTYRAMINE AND BILE DIVERSION LOWER THE AMINOPEPTIDASE ACTIVITY IN THE INTESTINAL BRUSH-BORDER MEMBRANE OF RATS

  K SONOYAMA, S KIRIYAMA, R NIKI

  JOURNAL OF NUTRITIONAL SCIENCE AND VITAMINOLOGY 39 (6) 617 - 625 0301-4800 1993/12 [Refereed][Not invited]
   

  To examine the effect of bile acids on the activity of intestinal aminopeptidase in vivo, we measured the activity of aminopeptidase in the intestinal mucosa from rats fed the diet containing cholestyramine which sequesters luminal bile acids (experiment 1) and from bile diverted rats (experiment 2). After 32 h fasting, rats were refed for 16 h either of a standard diet (25% casein diets), the same diet containing cholestyramine, or the fat-free diet in experiment 1. In the intestinal washing, the content of total bile acids was markedly decreased with feeding cholestyramine and activities of trypsin and chymotrypsin were also lowered with cholestyramine. Cholestyramine feeding decreased the specific activity of aminopeptidase in the homogenate of intestinal mucosa but increased the specific activities of sucrase and alkaline phosphatase. All these parameters were not modified by the fat-free diet. In experiment 2, bile diverted and sham operated rats were refed the standard diet for 16 h with prior 32 h fasting. Bile diversion, like cholestyramine feeding, lowered the content of total bile acids, the activities of pancreatic hydrolases in the intestinal washings, and the specific activity of aminopeptidase in the intestinal mucosa. The specific activity of sucrase in the intestinal mucosa was higher in bile diverted rats but the activity of alkaline phosphatase was not changed. These data indicate that the decreased abundance of intraluminal bile acid affects the activity of intestinal aminopeptidase not through the decreased absorption of dietary lipid. We propose that the intraluminal bile acids may be important for maintaining the activity of aminopeptidase while the degradation of sucrase by the pancreatic proteinases may be accelerated by the bile acids.
• FACILE PREPARATION OF RAT INTESTINAL-MUCOSA FOR ASSAY OF MUCOSAL ENZYME-ACTIVITY

  T KASAI, T TANAKA, S KIRIYAMA, K SONOYAMA

  JOURNAL OF NUTRITIONAL SCIENCE AND VITAMINOLOGY 39 (4) 399 - 403 0301-4800 1993/08 [Refereed][Not invited]
   

  Intestinal mucosa of rats was prepared by squeezing the frozen and thawed intestine. The method was much easier compared to the conventional method in which intestine was cut longitudinally or everted and mucosa was scraped. Jejunal mucosal weight prepared by the two methods was not different, but ileal mucosal weight prepared by squeezing was significantly heavier than that by scraped. No significant difference was observed between activity of some brush border enzymes (arylamidase, aminopeptidase, alkaline phosphatase, and sucrase) in mucosa prepared by squeezing and that by conventional scraping method in jejunum. Activity of brush border aminopeptidase and sucrase in ileal mucosa prepared by squeezing was significantly higher than that prepared by scraping.
• METHIONINE AND SOME METHIONINE DERIVATIVES ON GROWTH AND LIVER LIPIDS IN RATS FED A LOW CASEIN DIET - CORRELATION OF SUPPLEMENTAL EFFECTS WITH ABSORPTION RATES

  T KASAI, K SONOYAMA, S KIRIYAMA

  NIPPON NOGEIKAGAKU KAISHI-JOURNAL OF THE JAPAN SOCIETY FOR BIOSCIENCE BIOTECHNOLOGY AND AGROCHEMISTRY 社団法人 日本農芸化学会 61 (11) 1425 - 1434 0002-1407 1987/11 [Refereed][Not invited]
   

  Rats were fed an 8% casein diet supplemented with L-, D-methionine, LL- or DD-dimethionine. There were six groups in Experiment 1: 8% casein, 8% casein+0. 2% L-, D-methionine, LL-, DD-dimethionine or oligo-L-methionine (a mixture of hexamer and heptamer). There were seven groups in Experiment 2:8% casein, 8% casein+0.06, 0.08 or 1. 0% LL- or DD-dimethionine. In our previous experiment, fatty liver developed in rats fed an 8% casein diet supplemented with 0.2% L-methionine, but not in rats fed 8% casein with oligo-L-methionine. Since oligo-L-methionine hardly dissolves in water and is hydrol...
• DIFFERENCES IN ABSORPTION RATES OF 4 DIASTEREOISOMERS OF DIMETHIONINE BY RAT SMALL-INTESTINE

  T KASAI, K NISHINO, K SONOYAMA, S KIRIYAMA

  AGRICULTURAL AND BIOLOGICAL CHEMISTRY 50 (8) 2033 - 2037 0002-1369 1986/08 [Refereed][Not invited]
  

Books etc

• 花粉症等アレルギー疾患予防食品の開発

  (Contributorオリゴ糖の抗アレルギー効果)
  シーエムシー出版 2021
• 腸内細菌―宿主のクロストークと食事要因

  (Joint editor腸内細菌の健康機能を媒介する細胞外小胞―エクソソーム)
  2019
• 腸内細菌・口腔内細菌と全身疾患

  (Contributorメタボリックシンドローム)
  シーエムシー出版 2015
• 食物アレルギーの現状とリスク低減化食品素材の開発

  (Contributorオリゴ糖)
  シーエムシー出版 2015
• 食品機能性成分の吸収・代謝機構

  (Contributorオリゴ糖)
  シーエムシー出版 2013
• Handbook of diet, nutrition and the skin

  (ContributorFructo-oligosaccharides and skin inflammation)
  Wageningen Academic Publishers 2012
• ルミナコイド研究のフロンティア

  (Contributorルミナコイドによる腸内細菌叢の修飾とアレルギー予防)
  建帛社 2010

Conference Activities & Talks

• Exosome, a possible mediator of crosstalk between gut microbiota and host  [Invited]

  In Symposium 37 Exosome & microRNA, 7th International Conference on Food Factors  2019/12
• マウスの腸管オルガノイドを用いた腸上皮細胞機能の解析  [Invited]

  第104回日本栄養・食糧学会関東支部大会シンポジウム  2019/09
• 腸内細菌と宿主のクロストークを媒介するエクソソーム  [Invited]

  日本農芸化学会2019年度大会シンポジウム  2019/03
• プレ・プロバイオティクス研究から腸内細菌叢と宿主のクロストークを理解する  [Invited]

  第75回日本栄養・食糧学会中部支部大会特別講演  2018/12
• 腸内細菌叢と宿主の新しいクロストークシステムの発見をめざして  [Invited]

  2018年度 日本乳酸菌学会泊まり込みセミナー  2018/05
• 腸内細菌叢の健康機能を媒介する細胞外小胞  [Invited]

  日本栄養・食糧学会第72回大会シンポジウム  2018/05
• 腸管オルガノイドを用いたmicroRNAの解析  [Invited]

  日本農芸化学会2018年度大会シンポジウム  2018/03
• アレルギーの危険因子としての消化管常在真菌  [Invited]

  日本アレルギー学会第66回学術大会シンポジウム  2017/06
• 腸内細菌叢の情報を宿主に伝える血中エキソソームのmiRNA  [Invited]

  日本栄養・食糧学会第70回大会シンポジウム  2016/05
• 腸管オルガノイドを用いた腸粘膜上皮セロトニンシステムの解析  [Invited]

  日本農芸化学会2016年度大会シンポジウム  2016/03
• Communication tools mediating the health-promoting action of prebiotics and probiotics  [Invited]

  In Symposium 11 Immune Regulation by Gut Microbiota, Probiotics, and Prebiotics, 12th Asian Congress of Nutrition  2015/05
• 肥満，食餌および腸内細菌叢の関係におけるAkkermansia muciniphilaの意義  [Invited]

  第35回日本肥満学会 シンポジウム  2014/10
• メタボリック・シンドロームとプロバイオティックス  [Invited]

  日本臨床栄養学会 特別セミナー in 札幌  2014/07
• 腸粘膜バリアの維持・変容・破綻における腸内ムチン分解細菌の意義と難消化性糖類による修飾  [Invited]

  日本栄養・食糧学会第68回大会シンポジウム  2014/06
• 腸内細菌叢とメタボリックシンドローム  [Invited]

  日本細菌学会第86回総会シンポジウム  2013/03
• 高脂肪食がプロバイオティクスに与える消化管内ストレスとプレバイオティクスによるストレス対処  [Invited]

  日本食物繊維学会第16回学術集会シンポジウム  2011/11
• Prevention of murine contact hypersensitivity by prebiotics  [Invited]

  International Symposium at the 7th Annual Meeting of Japanese Association for Food Immunology  2011/10
• Prevention of allergic disease by use of prebiotics – animal studies  [Invited]

  International Symposium at Annual Meeting of Korean Society of Food Science and Technology  2010/06
• Prevention of allergic disease by prebiotics – animal studies  [Invited]

  日本農芸化学会2010年度大会シンポジウム  2010/03
• 難消化性糖類のアレルギー抑制作用  [Invited]

  第63回日本栄養・食糧学会大会シンポジウム  2009/05
• 難消化性糖類による腸内細菌叢の制御を介したアレルギー疾患予防の試み  [Invited]

  日本食物繊維学会第13回学術集会シンポジウム  2008/11
• 難消化性糖類による腸内細菌叢の制御を介したアレルギー疾患予防の試み  [Invited]

  日本農芸化学会北海道支部合同学術講演会シンポジウム  2008/11
• 食物アレルギーの増悪因子としての常在真菌Candida albicansと食餌によるその制御  [Invited]

  日本栄養・食糧学会中部支部大会シンポジウム  2007/07
• Is gastrointestinal colonization by Candida albicans a risk factor for food allergy?  [Invited]

  Second International Biomicrocosmos Workshop for the Enhancement of Gastro-Intestinal Biosphere Research  2006/02
• 食物アレルギー研究のための新しい消化管カンジダ症マウス  [Invited]

  日本食品免疫学会第1回学術大会シンポジウム  2005/11
• 食物繊維による腸管免疫の修飾  [Invited]

  Hindgut Club 第３回セミナー  1997/12
• 食品化学からみた食物繊維の作用機構－ビート食物繊維を例として  [Invited]

  第19回日本臨床栄養学会総会シンポジウム  1997/11

MISC

• 腸内細菌はマウスの大腸粘膜固有層白血球におけるmiR-200/BCL11B・ETS-1軸を介してIL-2産生を抑制する

  逢坂 文那, 唐津 勇吾, 門田 吉弘, 栃尾 巧, 園山 慶  日本栄養・食糧学会大会講演要旨集  75回-  111  -111  2021/07
• Identification of bifidobacterial genes that contribute to survival and colonization in the gastrointestinal tract of conventional mice using insertion-sequencing.

  山口颯人, 倉持碧海, 後藤恭宏, 小椋義俊, 園山慶, 林哲也, 横田篤, 吹谷智  日本乳酸菌学会誌  32-  (2)  2021
• 腸内細菌移植後の腸内細菌叢は1-ケストース摂取による小腸RegIII β発現を増加させる

  Teranart Udomsopagit, 新堀 絵美子, 門田 吉弘, 栃尾 巧, 園山 慶  日本栄養・食糧学会大会講演要旨集  74回-  202  -202  2020/04
• 大腸粘膜固有層単核球のmicroRNAの発現に影響を与える腸内細菌因子の探索

  逢坂 文那, 門田 吉弘, 栃尾 巧, 園山 慶  日本栄養・食糧学会大会講演要旨集  74回-  285  -285  2020/04
• 高脂肪食の摂取にともなう腸管IgAの腸内細菌に対する反応性の低下と肥満病態の関係性

  鶴田剛司, 園山慶, 西野直樹  日本栄養・食糧学会大会講演要旨集  74th-  2020
• 1-ケストース摂取により変化した大腸粘膜固有層単核球のmicroRNAの標的mRNAの解析

  逢坂 文那, 門田 吉弘, 栃尾 巧, 園山 慶  日本栄養・食糧学会大会講演要旨集  73回-  214  -214  2019/04
• Reactivity of secretory immunoglobulin A against gut bacteria is attenuated by high-fat diet consumption

  鶴田剛司, MUHOMAH Teresia Aluoch, 園山慶, 西野直樹  岡山実験動物研究会報  (35)  2019
• 組換え型REG3βの投与は食餌誘導性肥満マウスの末梢組織炎症およびインスリン抵抗性を緩和する

  鶴田剛司, 西野直樹, 園山慶  日本栄養・食糧学会大会講演要旨集  73rd-  2019
• 腸内細菌と宿主のクロストークを媒介するエクソソーム

  園山慶, 青木(吉田)綾子, 鶴田剛司  日本農芸化学会大会講演要旨集(Web)  2019-  2019
• 高脂肪食またはコール酸摂取マウス腸内における胆汁酸組成および細菌叢構成の動態解析による胆汁酸仮説の検証

  坂本佳奈子, 相磯知里, 松浦史佳, 吹谷智, 園山慶, 小椋義俊, 林哲也, 横田篤  日本農芸化学会大会講演要旨集(Web)  2019-  2019
• 1-ケストースを摂取させたマウス結腸におけるセロトニン含量とBifidobacteriaとの関係

  立岡 三沙, 逢坂 文那, Teranart Udomsopagit, 門田 吉弘, 栃尾 巧, 園山 慶  日本栄養・食糧学会大会講演要旨集  72回-  272  -272  2018/04
• 1-ケストースの摂取はマウス大腸粘膜固有層単核球のmiRNAプロファイルを変化させる

  逢坂 文那, 門田 吉弘, 栃尾 巧, 園山 慶  日本栄養・食糧学会大会講演要旨集  72回-  272  -272  2018/04
• マクロファージが分泌する細胞外小胞は乳酸菌の抗肥満効果を媒介する

  伊藤洋平, 鶴田剛司, 西野直樹, MUHOMAH Terry, 園山慶  岡山実験動物研究会報  (34)  2018
• 腸内細菌の健康機能を媒介する細胞外小胞

  園山慶, 鶴田剛司, 青木(吉田)綾子  日本栄養・食糧学会大会講演要旨集  72nd-  2018
• 乳酸菌を貪食したマクロファージが放出する細胞外小胞の脂肪細胞および骨格筋細胞への取り込み

  鶴田剛司, 伊藤洋平, 西野直樹, MUHOMAH Teresia Aluoch, 勝又恵実子, 園山慶  岡山実験動物研究会報  (34)  2018
• Lactobacillus plantarum No.14株を貪食したマクロファージが放出する細胞外小胞は内臓脂肪組織の脂肪蓄積抑制を媒介する

  伊藤洋平, 鶴田剛司, 西野直樹, 園山慶  日本栄養・食糧学会大会講演要旨集  72nd-  2018
• Lactobacillus plantarum No.14株を貪食したマクロファージが放出する細胞外小胞はマウスの内臓脂肪蓄積を抑制する

  伊藤洋平, 鶴田剛司, 西野直樹, 園山慶  日本栄養・食糧学会中国・四国支部大会講演要旨集  51st-  2018
• 分泌型免疫グロブリンAの腸内細菌への結合活性は高脂肪食の摂取により弱まる

  MUHOMAH Teresia Aluoch, 西野直樹, 園山慶, 桑高佳穂, 鶴田剛司  日本栄養・食糧学会中国・四国支部大会講演要旨集  51st-  2018
• REG3βが食餌誘導性肥満およびインスリン抵抗性に及ぼす影響

  磯田康太朗, 鶴田剛司, 西野直樹, 園山慶  日本栄養・食糧学会大会講演要旨集  72nd-  2018
• 組換え型REG3βの投与は高脂肪食摂取マウスにおける代謝異常を抑制する

  磯田康太朗, 鶴田剛司, 西野直樹, 園山慶  日本栄養・食糧学会中国・四国支部大会講演要旨集  51st-  2018
• ヒト由来ビフィズス菌Bifidobacterium longum105-Aが通常飼育マウス腸内に定着可能な飼育試験系の確立

  嶋田みな, 川瀬陽平, 園山慶, 小椋義俊, 林哲也, 横田篤, 吹谷智  日本乳酸菌学会誌  29-  (2)  2018
• アレルギーの危険因子としての消化管常在真菌

  園山 慶  臨床免疫・アレルギー科  69-  489  -493  2018  [Not refereed][Invited]
  
• プロバイオティクスの健康機能を媒介する循環血中のエクソソーム — 腸内細菌叢と宿主の新たなクロストークシステムか？

  園山 慶  化学と生物  56-  5  -6  2018  [Not refereed][Invited]
  
• Lactobacillus plantarum No.14株を貪食したマクロファージが放出する細胞外小胞は細胞種選択的に細胞の栄養代謝に影響を及ぼす

  鶴田剛司, 伊藤洋平, MUHOMAH Terry, 西野直樹, 園山慶  日本栄養・食糧学会大会講演要旨集  71st-  2017
• 高脂肪食摂取によるマウス腸内の胆汁酸量の増加が腸内細菌叢構成に与える影響

  相磯知里, 松浦史佳, 吹谷智, 渡辺真通, 園山慶, 小椋義俊, 林哲也, 横田篤  日本乳酸菌学会誌  28-  (2)  2017
• コール酸・ケノデオキシコール酸・デオキシコール酸各投与マウスにおける盲腸内胆汁酸組成および菌叢構成の変化

  松浦史佳, 相磯知里, 吹谷智, 渡辺真通, 園山慶, 小椋義俊, 林哲也, 横田篤  日本乳酸菌学会誌  28-  (2)  2017
• 腸内細菌叢とアレルギー

  園山 慶  アレルギーの臨床  37-  243  -247  2017  [Not refereed][Invited]
  
• マウスの小腸粘膜上皮におけるペプチドYY産生細胞はセロトニン受容体を発現する

  濱田晃弘, 尾崎洋輔, 斉藤伸一, 鶴田剛司, 青木(吉田)綾子, 園山慶  日本栄養・食糧学会大会講演要旨集  70th-  2016
• Lactobacillus plantarum No.14株を貪食したマクロファージが放出する細胞外小胞は白色脂肪細胞の脂肪蓄積を抑制する

  鶴田剛司, 伊藤洋平, 西野直樹, MUHOMAH Terry, 園山慶  日本栄養・食糧学会中国・四国支部大会講演要旨集  49th-  2016
• 短鎖脂肪酸がマウス結腸オルガノイドのセロトニンシステムにおよぼす影響

  尾崎洋輔, 斉藤伸一, 濱田晃弘, 鶴田剛司, 園山慶  日本栄養・食糧学会大会講演要旨集  70th-  2016
• 短鎖脂肪酸がマウス結腸オルガノイドの腸内分泌細胞分化に及ぼす影響

  尾崎洋輔, 斉藤伸一, 濱田晃弘, 鶴田剛司, 園山慶  日本農芸化学会大会講演要旨集(Web)  2016-  2016
• 腸内細菌叢の情報を宿主に伝える血中エキソソームのmiRNA

  園山慶, 鶴田剛司, 青木(吉田)綾子  日本栄養・食糧学会大会講演要旨集  70th-  2016
• 短期間の高脂肪食摂取によってマウスの腹腔内脂肪におけるリポ多糖結合タンパクの発現が増加する

  関真実, 三輪秋穂, 鶴田剛司, 園山慶  日本農芸化学会大会講演要旨集(Web)  2016-  2016
• プロバイオティクスを用いた肥満・メタボリックシンドロームの予防

  SONOYAMA Kei  The Lipid  27-  172  -178  2016  [Not refereed][Invited]
  
• マウス小腸陰窩オルガノイドにおけるセロトニン自己分泌刺激

  濱田晃弘, 鶴田剛司, 青木(吉田)綾子, 園山慶  日本農芸化学会大会講演要旨集(Web)  2015-  2015
• R-spondinおよびNogginの発現細胞株の作成とマウス腸管オルガノイドへの応用

  尾崎洋輔, 鶴田剛司, 斉藤伸一, 園山慶  日本農芸化学会北海道支部講演会講演要旨  2015-  2015
• フルクタン摂取時の盲腸ムチン濃度上昇へのムチン分解細菌Akkermansia muciniphilaの関与

  伊藤 謙, 源田 知美, 近藤 位旨, 日野 真吾, 園山 慶, 森田 達也  日本栄養・食糧学会大会講演要旨集  68回-  161  -161  2014/04  [Not refereed][Not invited]
  
• プロバイオティクスの炎症抑制作用を媒介するエキソソーム

  青木(吉田)綾子, 大角有里沙, 鶴田剛司, 園山慶  日本農芸化学会大会講演要旨集(Web)  2014-  2014
• 血清エキソソームはフラクトオリゴ糖の炎症抑制作用を媒介するか?

  角田妃菜子, 大角有里沙, 三輪明穂, 青木(吉田)綾子, 鶴田剛司, 園山慶  日本栄養・食糧学会大会講演要旨集  68th-  2014
• 肥満克服のターゲットとしての腸内細菌叢、「肥満克服のサイエンスー代謝の変容と脂肪細胞の制御から、最新治療まで」

  園山 慶  実験医学  32-  392  -398  2014  [Not refereed][Invited]
  
• Relationship between Gut Microbiota and Allergic Diseases and Its Modulation by Indigestible Oligosaccharides (2011's JDF Award for Excellence in Research)

  園山 慶  ルミナコイド研究  16-  (1)  1  -8  2012  [Not refereed][Invited]
  
• 肥満、「腸管と免疫・栄養ー腸内細菌から疾患を理解する」

  園山 慶  臨床栄養（臨時増刊）  120-  730  -735  2012  [Not refereed][Invited]
  
• 肥満の環境要因としての腸内細菌叢 : 肥満を予防・改善できるプロバイオティクスはあるか?

  園山 慶  化学と生物  49-  (10)  664  -665  2011/10  [Not refereed][Invited]
  
• 消化管常在真菌 Candida albicans はアレルギーのリスクファクターか? : 経口抗原に対する感作を促進. 炎症の増悪にも関わり

  園山 慶  化学と生物  49-  (7)  445  -447  2011/07  [Not refereed][Invited]
  
• Metabolic Syndrome and Gut Microbiota

  SONOYAMA Kei  Journal of Intestinal Microbiology  24-  (3)  193  -201  2010/07  [Not refereed][Invited]
   

  Increasing prevalence of obesity and associated metabolic disorders, such as diabetes and metabolic syndrome, is of global concern. Recent studies have focused on gut microbiota as an environmental factor that contributes to obesity and metabolic syndrome. Experimental studies using germ-free mice suggest that the gut microbiota affects nutrient acquisition and energy regulation in the host. The composition of gut microbiota has also been shown to differ in lean versus obese mice and humans. Furthermore, lipopolysaccharide derived from intestinal Gram-negative bacteria may act as a triggeri...
• 難消化性オリゴ糖によるアレルギーの予防・改善 (特集 アレルギー疾患に対する機能性食品)

  園山 慶  アレルギ-・免疫  15-  (5)  636  -643  2008/05  [Not refereed][Invited]
  
• Functions and Applications of α-Linked Galactooligosaccharide

  HASHIMOTO Hiroyuki, SONOYAMA Kei  KAGAKU TO SEIBUTSU  43-  (7)  431  -438  2005/07  [Not refereed][Invited]
  
• 肥満細胞は消化管カンジダ症による食物アレルゲンの透過こう進に関与する

  杉田竜介, 山口奈津, 三木篤子, 川端潤, 園山慶  日本農芸化学会北海道支部・日本土壌肥料学会北海道支部・日本生物工学会北日本支部・日本応用糖質科学会北海道支部・北海道農芸化学協会合同学術講演会講演要旨  2005-  2005
• 消化管カンジダ症がNC/Ngaマウスのアトピー性皮膚炎様症状に及ぼす影響

  山口奈津, 杉田竜介, 川端潤, 園山慶  日本農芸化学会大会講演要旨集  2005-  2005
• Enzymatic Synthesis of α-Linked Galactooligosaccharide (α-GOS) and Its Functions

  HASHIMOTO Hiroyuki, YAMASHITA Akiko, FUJITA Koki, OKADA Masamichi, MORI Shigeharu, SONOYAMA Kei, KITAHATA Sumio  Journal of applied glycoscience  51-  (2)  169  -176  2004/04/20  [Not refereed][Not invited]
  
• ひ細胞の抗原刺激によるサイトカイン産生にオリゴ糖が及ぼす影響

  斎藤亮, 園山慶, 山口奈津, 山下亜希子, 橋本博之, 森茂治, 岡田正通, 藤田孝輝, 菊地恵理子  日本農芸化学会大会講演要旨集  2004-  2004
• ケフィアによるオバルブミン(OVA)の腸管吸収抑制効果

  園山慶, 梅田智里, 山口奈津, 明石啓子, 元島英雅, 川端潤  日本栄養・食糧学会総会講演要旨集  58th-  2004
• 腸管カンジダ症モデルの作成と解析

  山口奈津, 園山慶, 名倉泰三, 仙波美博, 川端潤  日本栄養・食糧学会総会講演要旨集  57th-  2003
• 腸内カンジダ増殖が抗原の腸管吸収に及ぼす影響

  山口奈津, 園山慶, 名倉泰三, 仙波美博, 川端潤  日本農芸化学会大会講演要旨集  2003-  2003
• 酵素分解による小麦粉の低アレルゲン化

  渡辺 道子, 渡辺 純, 園山 慶, 田辺 創一  日本農芸化学会誌  76-  (11)  1090  -1091  2002/11/01  [Not refereed][Not invited]
  
• 大腸ガンは酪酸に対して耐性を獲得するのか? : ガン進展を抑制するのか, はたまた促進するのか

  園山 慶  化学と生物  40-  (11)  702  -703  2002/11  [Not refereed][Invited]
  
• ラフィノースが肥満細胞機能に及ぼす影響

  山口奈津, 葛西隆則, 園山慶  日本栄養・食糧学会総会講演要旨集  56th-  2002
• 食餌ラフィノースのアレルギー性ぜん息改善作用における腸内細菌の意義

  渡辺斉志, 渡辺純, 福本京香, 山口奈津, 葛西隆則, 園山慶  日本農芸化学会大会講演要旨集  2002-  2002
• 7. 定量的RT-PCR解析を用いたコレステロール負荷ラットにおけるn-6およびn-3脂肪酸食の長期投与がコレステロール代謝に関連するmRNA発現に及ぼす影響

  福島 道広, 島田 謙一郎, 大橋 哲, 斉藤 秀俊, 園山 慶, 関川 三男, 中野 益男  ビタミン  75-  (8)  445  -446  2001/08/25  [Not refereed][Not invited]
  
• Food Science Challenges Intractable Wheat Allergy

  TANABE Soichi, WATANABE Jun, SONOYAMA Kei, WATANABE Michiko  KAGAKU TO SEIBUTSU  39-  (7)  440  -447  2001/07/25  [Not refereed][Not invited]
  
• Regulation of Apolipoprotein Expression in Intestine

  SONOYAMA Kei  Nippon Eiyo Shokuryo Gakkaishi  53-  (2)  65  -70  2000/04  [Not refereed][Invited]
   

  Apolipoproteins play a central role in transport and metabolism of lipids in animals. We investigated the regulatory mechanisms responsible for expression of apolipoproteins in the small intestine. Animal experiments using surgical procedures suggested that the biliary component is a luminal factor regulating apolipoprotein A-IV expression in the intestine. In addition, peptide YY stimulated the apolipoprotein A-IV expression in Caco-2 intestinal cells in a dose- and time-dependent manner, suggesting that peptide YY is a humoral factor regulating the expression of apolipoprotein A-IV. Furth...
• 食品化学からみた食物繊維の作用機構--ビ-ト食物繊維を例として (第19回日本臨床栄養学会総会) -- (シンポジウム(2)食物繊維の栄養機能と臨床への応用)

  園山 慶  日本臨床栄養学会雑誌  20-  (2)  35  -39  1998/06  [Not refereed][Invited]
  
• 食物繊維による血漿コレステロール低下作用のメカニズムを探る 各種臓器でのアポリポ蛋白遺伝子の発現と相関

  園山 慶  化学と生物  33-  (7)  418  -419  1995/07  [Not refereed][Invited]
  

Awards & Honors

• 2011 日本食物繊維学会学会賞
  
• 2011 日本食品免疫学会学会賞
  
• 2001 Biosci. Biotechnol. Biochem. 論文賞
  
• 1999 日本栄養・食糧学会奨励賞
  
• 1999 May, 1999 Young Scientist Award (The Japan Society of Nutrition and Food Science)March, 2001 Paper Award in Biosci. Biotechnol. Biochem. (Japan Society for Bioscience, Biotechnology and Agrochemistry)
  

Research Grants & Projects

• 腸管免疫のホメオスタシスに寄与するmicroRNAと食事/腸内細菌叢の要因

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2020/04 -2023/03 

  Author : 園山 慶

   

  本研究では、「miRNAによる遺伝子サイレンシングを介したT細胞分化の調節は、腸内細菌叢が腸管免疫のホメオスタシスに寄与する分子基盤である」という仮説を立て、以下のように検証を進めた。 1) 腸内細菌叢の存在がマウス大腸の粘膜固有層白血球におけるmiR-200ファミリーメンバーの発現を増加させ、それらの標的遺伝子のサイレンシングを介して腸管免疫の調節に寄与するという前年の実績に基づき、このことをより直接的に証明した。すなわち、培養細胞にmiR-200ファミリーメンバーを遺伝子導入した際の標的遺伝子のサイレンシングを明らかにした。 2) 難消化性オリゴ糖を摂取させたマウスの大腸粘膜固有層白血球において、miR-200ファミリーメンバーの発現が増加し、このことに腸内に常在するビフィズス菌が寄与することを示唆する知見を得た。すなわち、腸内細菌叢のメタ16S解析の結果、ビフィズス菌の存在量とmiR-200ファミリーメンバーの発現との関連が示された。 以上のように、本年度の研究により、腸内細菌叢の構成を変化させるような食品が腸管免疫に影響を及ぼす際に、microRNAによる遺伝子サイレンシングが重要な役割を果たすことを明らかにした。
• Circulating microRNA mediating food-gut microbiota-host linkage

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2017/04 -2020/03 

  Author : Sonoyama Kei

   

  We found that indigestible oligosaccharides increase the expression of a group of microRNAs (miRNAs) in colonic lamina propria lymphocytes of mice. Many of these miRNAs were consistent with miRNAs whose expression levels were higher in conventional mice than in germ-free mice. These findings suggest that miRNA-induced gene silencing is involved in the gut microbiota influence on gut immune homeostasis. Additionally, by transfecting the miRNA mimics to a human T-cell line Jurkat, we established an experimental system to search for target genes and to examine the effect on T-cell differentiation.
• Identification and functional analysis of genes important for the activity of bifidobacteria in the intestine harboring intestinal microbiota

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

  Date (from‐to) : 2016/04 -2019/03 

  Author : Fukiya Satoru

   

  Bifidobacteria are well-known for their beneficial effects on human health, but the molecular mechanisms how bifidobacteria are colonizing and surviving in the intestine are not fully understood especially in the presence of the intestinal microbiota. To clarify the molecular mechanisms, the breeding system of the conventional mice that allows long-term colonization of the human-derived bifidobacteria was established. Using this system, bifidobacterial genes specifically expressed in the intestine were identified by R-IVET analysis. Furthermore, a transposon mutant library comprising 48,000 mutants was constructed and administered to the germ-free mice. Increase and decrease of each mutant in the intestine were comprehensively evaluated by the INSeq analysis. Finally, 442 genes were identified that were important to the colonization and survival in the intestine.
• プレ/プロバイオティクスの健康機能を生体内で媒介する細胞分子基盤の解析

  文部科学省：科学研究費補助金（基盤研究B）

  Date (from‐to) : 2014/04 -2017/03 

  Author : 園山 慶
• 冬眠動物における腸粘膜上皮の細胞更新遅滞現象

  文部科学省：科学研究費補助金（挑戦的萌芽研究）

  Date (from‐to) : 2013/04 -2015/03 

  Author : 園山 慶
• Studies on the development of anti-obesity synbiotics

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2011 -2012 

  Author : SONOYAMA Kei

   

  We previously showed that Lactobacillus plantarum No.14 reduces obesity in mice, whereas the cellular and molecular mechanisms remain unknown. The present study demonstrated that L. plantarum No.14 reduces obesity, adiopose tissue inflammation, and insulin resistance in type 2 deiabetic mice. In addition, we showed that exosomes isolated from the serum of mice fed L. plantarum No.14 reduce the production of inflammatory cytokines in peritoneal exudate cells and murine macrophage cell line RAW264.7. The results suggest that exosomes play a role in cell-to-cell communication in terms of anti-inflammatory action of L. plantarum No.14.
• Gut microbiota and gut mucosal barrier in hibernating animals

  Japan Society for the Promotion of Science：Grants-in-Aid for Scientific Research

  Date (from‐to) : 2011 -2011 

  Author : SONOYAMA Kei, TSUBOTA Toshio, TAKIGUCHI Mitsuyoshi

   

  This study investigated the control of gut microbiota and gut mucosal barrier in hibernating animals. In hibernating Syrian hamsters, we observed higher concentrations of secretory antibodies in the small intestinal contents, suggesting its contribution to protection against infection. Indeed, no bacteria were observed in the mesenteric lymph node of hibernating hamsters. Analysis of lymphocyte profiles in the immune tissues indicated that ability to protect against infection and to regulate immune function may be maintained in the gut of hibernating hamsters. Furthermore, we found that cell renewal of intestinal epithelium is markedly retarded in hibernating hamsters.
• Studies on food prevention of immune diseases through modulationof gut microbiota

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(B))

  Date (from‐to) : 2007 -2008 

  Author : Kei SONOYAMA, Jun WATANABE

   

  腸内細菌は免疫系に影響するので、難消化性糖類などの食品によって腸内細菌叢を修飾することにより、アレルギーなどの免疫疾患を予防・治療できるかもしれない。このことを動物実験によって調べた結果、授乳期の仔マウスの腸内細菌叢を修飾することによって発育後のアレルギー性皮膚炎を改善できること、米品種が腸内細菌叢を修飾するとともに食物アレルギーにも影響を及ぼすこと、消化管内のカンジダ菌がアレルギーの危険因子となりうることを示すことができた
• 冬眠動物から探る新しい腸管粘膜バリア・粘膜免疫機構と食餌によるその制御

  文部科学省：科学研究費補助金(萌芽研究)

  Date (from‐to) : 2007 -2008 

  Author : 園山 慶, 渡辺 純, 間野 勉, 富沢 昌章, 釣賀 一二三

   

  冬眠動物における腸内細菌と宿主の腸管粘膜バリアおよび免疫会系との関係について、以下の三点を検討した。1. シリアンハムスターの冬眠個体、覚醒期の自由摂食個体および絶食個体の盲腸内細菌叢を16SrRNA遺伝子のクローンライブラリを構築して解析した結果、冬眠個体および自由摂食個体ではClostridiaが優占するが、絶食個体ではムチン分解菌であるAkkermansia muciniphilaが著しく増加することを明らかにした。また冬眠個体では覚醒期自由摂食個体と比して総菌数は変化しないにもかかわらず盲腸内容物中の短鎖脂肪酸濃度が低下していたので、腸内細菌も低温のために休眠状態となっていることが示唆された。2. シリアンハムスターの冬眠個体において、腸管粘膜の肥厚が生じることを平成19年度の研究で示したが、その機構について粘膜上皮の細胞回転に着目して免疫組織化学的に解析した結果、細胞回転の著しい遅延が生じていることが明らかとなり、このことが粘膜肥厚に関与することが示唆された。3. 平成19年度の研究において取得した抗ヒグマIgA抗体を用いて、夏季の有害鳥獣駆除により捕獲された野生エゾヒグマの腸管の免疫組織化学的解析を行い、IgA産生細胞の組織分布様式を明らかにし、このことからヒグマの腸管パイエル板が免疫応答を行う二次リンパ組織であることを示唆した.
• Design of immune tolerance inducing peptides and analysis of their allergy-suppressive effects

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(若手研究(B))

  Date (from‐to) : 2006 -2008 

  Author : Jun WATANABE, Kei SONOYAMA, Susumu ITO, Hidenori TAGUCHI

   

  免疫寛容誘導物質の作製においては、低アレルゲン化小麦粉水溶性画分が免疫寛容誘導活性をもつことを明らかとし、活性本体のペプチドの単離を試みた。腸内細菌とアレルギー発症との関わりを調べ、Candida albicans の定着や幼少期の抗生物質投与による菌叢変化がアレルギーの増悪に関わること、難消化性オリゴ糖の摂取は菌叢変化を介してアレルギー発症に抑制的に作用することを示した。
• Cellular and molecular mechanisms for suppression of allergic response by dietary indigestible oligosaccharides

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(B))

  Date (from‐to) : 2004 -2006 

  Author : Kei SONOYAMA, 福島 道広

   

  We aimed to clarify cellular and molecular mechanisms for suppression of allergic response by α-galactooligosaccharides (α-GOS).1. Dietary α-GOS did not reduce Con A-induced hepatitis and endotoxin shock in mice. IFN-γ production by mouse splenocytes ex vivo was stimulated by LPS, antigen, and endogenouse and exogenous ligands for NKT cells. Because the IFN-γ production was inhibited by anti-CD1d antibody, CD1d-restricted activation of NKT cells is probably responsible for the IFN-γ production. However, supplementation of γ-GOS did not affect the IFN-γ production. These findings suggest tha...
• Effects of bean resistant starches and mushroom fibers on lipid metabolism in rats.

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(C))

  Date (from‐to) : 2003 -2004 

  Author : Michihiro FUKUSHIMA, 園山 慶, 関川 三男, 小嶋 道之, 島田 謙一郎

   

  1.The hypocholesterolemic effects in rats fed the enzyme-resistant fractions of adzuki, kintoki and tebou beans were evident compared with rats fed a cellulose diet. The enzyme-resistant fractions elevated fecal acidic steroid excretions, and reduced the serum total cholesterol, HDL-cholesterol, and VLDL + IDL + LDL-cholesterol concentrations. It appears that the cholesterol-lowering effect of enzyme-resistant fractions is due to the enhanced cholesterol 7α-hydroxylase mRNA level in the adzuk, kintoki and tebou groups, SR-B1 mRNA level in the adzuki group, and fecal steroid excretion accele...
• Role of activin A in induction of growth arrest and apoptosis by butyrate in colon cancer cells

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(C))

  Date (from‐to) : 2001 -2002 

  Author : Kei SONOYAMA

   

  1. Colorectal tumor-suppressing effect of dietary fiber has been reportedly associated with its fermentation product butyrate. The present study investigated whether activin A mediates butyrate-induced growth arrest and apoptosis in colon cancer cells.2. Butyrate induced G1 arrest and apoptosis in human colon cancer cell line HT-29. Butyrate also stimulated activin A gene in time-and dose-dependent manner. Because trichostain A also stimulated activin A gene, the effect of butyrate is possibly associated with the inhibition of histone acetylase. Stimulation of activin A gene by butyrate see...
• Studies on allergy-preventive effect of hypoallergenic wheat flour and indigestible oligosaccharide raffinose

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(B))

  Date (from‐to) : 2001 -2002 

  Author : Takanori KASAI, 渡辺 純, 園山 慶

   

  1. The present study investigated the mechanism by which hypoallergenic wheat flour and indigestible oligosaccharide raffinose prevent allergy.2. Hypoallergenic wheat flour prevented gluten-specific allergic airway eosinophilia in Brown Norway rats. Hypoallergenic wheat flour suppressed systemic immuneresponse against gluten, suggesting oral tolerance. Transfer of splenocytes from donor mice fed hypoallergenic wheat flour to recipient mice induced suppression of gluten-specific antibody levels in recipient mice. The results suggest that oral tolerance induced by hypoallergenic wheat flour i...
• Neuroendocrine regulation of expression of apolipoprotein A-IV gene in intestinal epithelial cells

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(C))

  Date (from‐to) : 1999 -2000 

  Author : Kei SONOYAMA

   

  The present study investigated the role of neural and endocrine factors in the regulation of expression of apolipoprotein A-IV (A-IV) gene in intestinal epithelial cells. Intravenous infusion of ganglionic blocker, cholinergic antagonist, and NOS inhibitor suppressed the expression of A-IV mRNA in rat ileum, suggesting regulation of A-IV gene by cholinergic and NOergic neurons. A-TV mRNA was not affected byβ-adrenergic, dopaminergic, serotonergic, ATRergic antagonists. Since ganglionic blocker did not suppress the increase in A-IV mRNA level by dietary lipid and in adaptive ileum after mass...
• アポリポタンパク質A-IV遺伝子発現を制御するラット小腸由来体液性因子の探索

  文部科学省：科学研究費補助金(奨励研究(A))

  Date (from‐to) : 1997 -1998 

  Author : 園山 慶

   

  小腸上皮細胞に発現するアポリポタンパク(アポ)A-IVは、脂質代謝ならびに食欲・消化管機能調節に関与するので、このものの発現調節機構を明らかにすることは脂質代謝、摂食行動、消化管機能の調節機構を解明することにもつながる。本研究では、小腸上皮細胞におけるアポA-IV遺伝子発現に関与する小腸由来体液性因子について主として細胞培養系を用いて解析した。とりわけ筆者のこれまでのラットを用いた解析により消化管ホルモンであるペプチドYY(PYY)がアポA-IV発現に関与することが予想されたので、このものがアポA-IV遺伝子発現に及ぼす影響を解析した。小腸上皮細胞様に分化する事が知られているヒト結腸ガン由来細胞株Caco-2をモデルとして用い、PYYがアポA-IVmRNAレベルに及ぼす影響について、半定量的なRT-PCR及びサザン解析により検討した。その結果、分化したCaco-2細胞において、PYYは用量ならびに時間依存性にアポA-IVmRNAレベルを増加させた。一方、小腸上皮細胞において発現することが知られている、アポA-I、B、及びC-III mRNAには変化が見られなかった。更に、PYYは細胞膜表面の特異的な受容体を介してその生理機能を発揮するが、本研究においてCaco-2細胞におけるPYY受容体のmRNA発現について解析したところ、Y1受容体のmRNAはPYY添加後、一過性の増加を示...
• Physiological and Biochemical Study of Gastrointestinal Functions.

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(A))

  Date (from‐to) : 1997 -1998 

  Author : Yoritaka AOYAMA, 石塚 敏, 園山 慶, 川端 潤, 葛西 隆則

   

  (1) When rats were fed an orotic acid diet, liver lipid level of rats fed a diet containing casein was significantly higher than that of a diet containing egg protein or soybean protein. The addition of an orotic acid to a diet containing casein caused to decrease liver superoxide dismutase activity, to increase serum ornithine carbamoyl transferase activity, to increase the hepatic concentration of 1,2-diacylglycerol. Liver damage by administration of an orotic acid might be due to the increased concentration of 1,2-diaclglycerol and decreased activity of hepatic superoxide dismutase.(2) 9...
• Role of genotoxic stress-induced apoptosis in the protective effect of dietary fiber against tumorigenesis in the large bowel

  Ministry of Education, Culture, Sports, Science and Technology：Grants-in-Aid for Scientific Research(基盤研究(C))

  Date (from‐to) : 1997 -1998 

  Author : Takanori KASAI, 石塚 敏, 園山 慶

   

  The present study investigated the role of genotoxic stress-induced apoptosis in the protective effect of dietary fiber against tumorigenesis in the large bowel. Dietary beet fiber increased the apoptotic frequency in rat colonic epithelium at 6 h after a subcutaneous injection of carcinogen 1 , 2-dimethylhydrazine (DM11). The finding suggests that increased efficiency of elimination of damaged cells may be associated with the anti-tumorigenic effect of dietary beet fiber. Semi-quantitative RT-PCR demonstrated that increase in the expression of cyclin-dependent kinase inhibitor p21 and apop...
• ラット小腸アポリポタンパクBm RNA editingに関する栄養生理学的解析

  文部科学省：科学研究費補助金(奨励研究(A))

  Date (from‐to) : 1996 -1996 

  Author : 園山 慶

   

  小腸に発現するアポリポタンパク(アポ)Bはカイロマイクロンの主要タンパクで食餌脂質の輸送に不可欠な因子であり、一方冠状動脈心疾患の発症との相関が見いだされている。血漿アポBは、B-48とB-100の2つの分子種からなり、これらは単一のアポB遺伝子からmRNA editingという機構により合成される。本研究では食餌による血漿コレステロール低下作用におけるアポB遺伝子発現動態の関与を明らかにすることを目的とし、更に小腸におけるアポBの遺伝子発現制御機構を解明する一環として、小腸切除後の残存回腸における応答を観察した。血漿コレステロール低下作用を有するビ-ト繊維を摂取させたラットの小腸及び肝臓アポ BmRNAをNorthern blottingにより分析したところ、無繊維飼料摂取ラットに比して低下傾向が見られた。アポB mRNAのediting siteを含む275bpのcDNAをRT-PCRにより取得し、それを鋳型にしてprimer extension/dideoxy termination assayを行うことによりアポB mRNA editingを比較したところ、ビ-ト繊維は肝臓及び小腸におけるアポ BmRNA editingに影響を及ぼさなかった。現在、血漿コレステロール低下作用を示す他のregimen(大豆タンパク質及び魚油添加飼料)が、アポB mRNAレベル及びアポB...
• アポリポプロテインA-I及びA-IV遺伝子発現調節機構に関する栄養生理学的解析

  文部科学省：科学研究費補助金(奨励研究(A))

  Date (from‐to) : 1995 -1995 

  Author : 園山 慶

   

  本研究ではこれまでに筆者が見いだした現象をもとに、小腸広範囲切除後に残存回腸のアポA-IV遺伝子発現を正に制御する胆汁成分を同定すること、並びに食物繊維の腸内発酵に由来して肝臓のアポA-I及びA-IVの遺伝子発現を制御する因子を同定することを目的として、以下の検討を行った。まず、intactな小腸を有するラットに対しても胆汁が回腸アポA-IV遺伝子発現を亢進させるか否かを検討した。即ち内因性の胆汁が空腸を迂回して回腸に流入するモデル、及び回腸にカテーテルを挿入して外因性に胆汁を注入するモデルを作成し、回腸アポA-IVmRNAを分析した。その結果、いずれのモデルにおいてもアポA-VmRNAレベルの顕著な増加が見られた。また後者のモデルを用いて胆汁酸溶液を注入したところ、アポA-IVmRNAに大きな変化は見られなかった。これらより、intactな小腸を有する動物においても回腸に胆汁が多量に流入することによりアポA-IV遺伝子発現の顕著な亢進が生じ、それは少なくとも胆汁酸吸収増加によっているのではないことが明らかになった。現在、その他の胆汁成分としてコレステロール、脂肪酸、リン脂質の影響を同様なモデルにより検討中である。また、小腸上皮細胞並びに肝細胞のin vitro系として各々Caco-2並びにHepG2細胞株の培養を行い、アポA-I及びA-IVmRNA、及び培養液中に分泌された...

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