研究者情報

マスター

アカウント（マスター）

• 氏名

  園山　慶(ソノヤマ　ケイ), ソノヤマ　ケイ

所属（マスター）

• 農学研究院　基盤研究部門　生物機能化学分野

所属（マスター）

• 農学研究院　基盤研究部門　生物機能化学分野

独自項目

syllabus

• 2021, 大学院共通授業科目（一般科目）：自然科学・応用科学, Inter-Graduate School Classes(General Subject):Natural and Applied Sciences, 修士課程, 大学院共通科目, 食品、消化管、食品機能、腸内細菌叢、生理活性
• 2021, 食品科学特論, Advanced Review of Food Science, 修士課程, 農学院, 食品、消化管、食品機能、腸内細菌叢、生理活性
• 2021, 食品科学特論演習, Advanced Seminar on Food Science, 修士課程, 農学院, 食品、消化管、食品機能、腸内細菌叢、生理活性
• 2021, 大学院共通授業科目（一般科目）：自然科学・応用科学, Inter-Graduate School Classes(General Subject):Natural and Applied Sciences, 修士課程, 大学院共通科目, 腸内細菌，腸内細菌叢，ルーメン，プロバイオティクス，乳酸菌，ビフィズス菌、腸管免疫、疾病発症 Gut microbes, Gut microbiota, Rumen, Probiotics, Lactic acid bacteria, Bifidobacteria, Gut immunity, Disease development
• 2021, 胃腸内圏微生物学特論, Advanced Gastrointestinal Microbiology, 修士課程, 農学院, 腸内細菌，腸内細菌叢，ルーメン，プロバイオティクス，乳酸菌，ビフィズス菌、腸管免疫、疾病発症 Gut microbes, Gut microbiota, Rumen, Probiotics, Lactic acid bacteria, Bifidobacteria, Gut immunity, Disease development
• 2021, 食品栄養化学概論, Introduction of Food and Nutritional Chemistry, 学士課程, 農学部, 消化管、栄養素、エネルギー代謝、運動、栄養学の歴史
• 2021, 環境と人間, Environment and People, 学士課程, 全学教育, 共生、寄生、進化、生物生産、植物、哺乳動物、昆虫、微生物、ウィルス、分子生物学、生態学、環境保全
• 2021, 一般教育演習(ﾌﾚｯｼｭﾏﾝｾﾐﾅｰ), Freshman Seminar, 学士課程, 全学教育, 食料、安全、作物生産、放射能汚染、食品微生物、酵素利用技術、毒物、ガン、アレルギー、メタボリックシンドローム、腸内細菌叢、保健機能食品
• 2021, 食品機能化学Ⅰ, Food Biochemistry I, 学士課程, 農学部, アレルギー、メタボリックシンドローム、腸内微生物、食品成分

researchmap

プロフィール情報

学位

• 博士（農学）（北海道大学）
• 農学修士（北海道大学）

プロフィール情報

• 姓

  園山, ソノヤマ

• 名

  慶, ケイ

• ID各種

  200901023144219900

対象リソース

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

業績リスト

研究キーワード

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

研究分野

• ライフサイエンス / 食品科学

経歴

• 2018年10月 - 現在 北海道大学大学院農学研究院 基盤研究部門生物機能化学分野食品機能化学研究室 教授
• 2007年04月 - 2018年09月 - 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年06月 - 2007年03月 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年 - 2007年 Associate Professor
• 1999年04月 - 2003年03月 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年 - 2003年 Research Associate
• 1992年04月 - 1999年03月 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年 - 1999年 Research Associate
• 1988年04月 - 1991年01月 ライオン株式会社生物科学研究所 研究員
• 1988年 - 1991年 Researcher

学歴

•         - 1992年   北海道大学   農学研究科   農芸化学専攻博士課程
•         - 1992年   北海道大学
•         - 1988年   北海道大学   農学研究科   農芸化学専攻修士課程
•         - 1988年   北海道大学
•         - 1986年   北海道大学   農学部   農芸化学科
•         - 1986年   北海道大学

委員歴

• 2008年04月 - 現在   日本食物繊維学会   理事   日本食物繊維学会
• 2006年11月 - 現在   日本栄養・食糧学会   参与   日本栄養・食糧学会
• 2004年10月 - 現在   日本食品免疫学会   幹事

受賞

• 2011年 日本食物繊維学会学会賞
  
• 2011年 日本食品免疫学会学会賞
  
• 2001年 Biosci. Biotechnol. Biochem. 論文賞
  
• 1999年 日本栄養・食糧学会奨励賞
  
• 1999年 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)
  

論文

• Unabsorbed Fecal Fat Content Correlates with a Reduction of Immunoglobulin a Coating of Gut Bacteria in High-Lard Diet-Fed Mice.

  Emiko Katsumata, Takeshi Tsuruta, Kei Sonoyama, Takashi Yoshida, Mio Sasaki, Mao Teraoka, Tianyang Wang, Naoki Nishino

  Molecular nutrition & food research 68 15 e2400078  2024年08月 

  SCOPE: Immunoglobulin A (IgA) selectively coats gut bacteria and contributes to regulatory functions in gastrointestinal inflammation and glucose metabolism. Excess intake of lard leads to decrease in the IgA coating of gut bacteria, although the underlying mechanisms remain unknown. This study validates how unabsorbed fat derived from a high-lard diet in the gut affects the IgA coating of bacteria, as assessed in mouse models using three types of dietary fat (lard, medium-, and long-chain triglycerides [MLCTs], and medium-chain triglycerides [MCTs]) exhibiting different digestibilities. METHODS AND RESULTS: C57BL/6J mice are maintained on diets containing lard, MLCTs, or MCTs at 7% or 30% w/w for 10 weeks (n = 6 per group). The fecal fatty acid concentration is measured to quantify unabsorbed fat content. The ratio of IgA-coated bacteria to total bacteria (IgA coating ratio) in the feces is measured by flow cytometry. Compared to lard-fed mice, MLCT- and MCT-fed mice exhibit lower fecal concentrations of palmitic acid, stearic acid, and oleic acid and higher IgA coating ratios at both 7% and 30% dietary fat, and these parameters exhibit significant negative correlations. CONCLUSION: Unabsorbed fat content in the gut may result in attenuated IgA coating of bacteria in high-lard diet-fed mice.
• 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 2023年12月
• 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年07月21日 

  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年07月 

  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 2023年04月30日
• 抗生剤およびビフィズス菌の投与がマウス大腸粘膜固有層白血球におけるmiR-200ファミリーの発現におよぼす影響

  本間 大貴, 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 77回 185 - 185 (公社)日本栄養・食糧学会 2023年03月
• 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年
• 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年
• 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年
• 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 2021年09月 [査読有り]
  
• 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 2021年04月14日 [査読有り]
   

  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 2021年01月 [査読有り]
   

  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 2020年10月 [査読有り]
   

  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年08月 [査読有り]
   

  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 74回 203 - 203 (公社)日本栄養・食糧学会 2020年04月
• 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年03月13日 [査読有り][通常論文]
   

  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 2018年09月02日 [査読有り]
   

  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 2018年06月 [査読有り]
  
• 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 72回 187 - 187 (公社)日本栄養・食糧学会 2018年04月
• 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 2017年10月 [査読有り][通常論文]
   

  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 2017年07月 [査読有り][通常論文]
   

  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.

  Tsuruta T, Saito S, Osaki Y, Hamada A, Aoki-Yoshida A, Sonoyama K

  Biochemical and biophysical research communications 474 1 161 - 167 2016年05月 [査読有り][通常論文]
   

  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 2016年 [査読有り][通常論文]
   

  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 2015年03月 [査読有り][通常論文]
   

  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.

  Asano M, Yamamoto T, Tsuruta T, Nishimura N, Sonoyama K

  Development, Growth & Differentiation 57 1 68 - 73 2015年01月 [査読有り][通常論文]
   

  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 2014年08月 [査読有り][通常論文]
   

  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 2014年06月 [査読有り][通常論文]
   

  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 2013年01月 [査読有り][通常論文]
   

  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 2013年 [査読有り][通常論文]
  
• 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 2012年09月 [査読有り][通常論文]
   

  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 2012年08月 [査読有り][通常論文]
   

  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 2012年 [査読有り][通常論文]
  
• 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 2012年 [査読有り][通常論文]
  
• 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 2012年 [査読有り][通常論文]
  
• 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 2012年 [査読有り][通常論文]
  
• 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 2011年05月 [査読有り][通常論文]
   

  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 2011年04月 [査読有り][通常論文]
   

  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 2010年12月 [査読有り][通常論文]
   

  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 2010年11月 [査読有り][通常論文]
   

  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 2010年08月 [査読有り][通常論文]
   

  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 2010年07月 [査読有り][通常論文]
   

  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 2010年02月 [査読有り][通常論文]
   

  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 2010年02月 [査読有り][通常論文]
   

  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 2010年02月 [査読有り][通常論文]
   

  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 2010年02月 [査読有り][通常論文]
   

  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 2010年01月 [査読有り][通常論文]
   

  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.
• 母マウスのフラクトオリゴ糖摂取は仔マウスの発育後の経口免疫寛容の誘導および腸管粘膜バリアに影響を及ぼさない

  吉田知世, 後藤悠, 武村直紀, 藤原麗子, 園山慶

  日本食物繊維学会誌 14 1 23 - 32 日本食物繊維学会 2010年 [査読有り][通常論文]
  
• 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 2009年10月 [査読有り][通常論文]
   

  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 2009年09月 [査読有り][通常論文]
   

  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年 [査読有り][通常論文]
  
• 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 2008年08月 [査読有り][通常論文]
   

  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 2008年07月 [査読有り][通常論文]
   

  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 2008年06月 [査読有り][通常論文]
   

  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 2006年07月 [査読有り][通常論文]
   

  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 2005年03月 [査読有り][通常論文]
   

  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 2005年01月 [査読有り][通常論文]
   

  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 2005年01月 [査読有り][通常論文]
   

  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 2004年08月 [査読有り][通常論文]
   

  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 2004年 [査読有り][通常論文]
   

  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 2004年01月 [査読有り][通常論文]
   

  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 2003年11月 [査読有り][通常論文]
   

  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 2003年09月 [査読有り][通常論文]
   

  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 2003年06月 [査読有り][通常論文]
   

  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 57回 73 - 73 (公社)日本栄養・食糧学会 2003年04月
• Isolation and structural elucidation of a peptide derived from edam cheese that inhibits β-lactoglobulin transport

  TANABE S, TESAKI S, WATANABE J, FUKUSHI E, SONOYAMA K, KAWABATA J

  Journal of Dairy Science 86 2 464 - 468 2003年02月 [査読有り][通常論文]
   

  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 2002年09月 [査読有り][通常論文]
   

  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 56回 119 - 119 (公社)日本栄養・食糧学会 2002年06月
• 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 2001年09月 [査読有り][通常論文]
   

  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 2001年09月 [査読有り][通常論文]
   

  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 2001年08月 [査読有り][通常論文]
   

  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", J. Nutr. Sci. Vitaminol. 47: 228-235 (20・・・

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

  Journal of Nutritional Science and Vitaminology 47 3 228 - 235 2001年06月 [査読有り][通常論文]
   

  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", J. Nutr. Sci. Vitaminol. 47: 228-235 (2001)*
• 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  2001年03月05日 [査読有り][通常論文]
  
• 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  2001年03月05日 [査読有り][通常論文]
  
• 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 2001年03月 [査読有り][通常論文]
   

  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 2001年02月 [査読有り][通常論文]
   

  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 2000年12月 [査読有り][通常論文]
   

  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 2000年09月 [査読有り][通常論文]
   

  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 2000年06月 [査読有り][通常論文]
   

  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 2000年03月 [査読有り][通常論文]
   

  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 2000年03月 [査読有り][通常論文]
   

  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 2000年01月 [査読有り][通常論文]
   

  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 1999年07月 [査読有り][通常論文]
   

  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 1997年11月 [査読有り][通常論文]
   

  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 1997年08月 [査読有り][通常論文]
   

  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 1997年05月 [査読有り][通常論文]
   

  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 70 0 30 - 30 社団法人日本農芸化学会 1996年03月05日 [査読有り][通常論文]
  
• 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 70 0 32 - 32 社団法人日本農芸化学会 1996年03月05日 [査読有り][通常論文]
  
• 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 1996年03月 [査読有り][通常論文]
   

  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 69 0 345 - 345 社団法人日本農芸化学会 1995年07月05日 [査読有り][通常論文]
  
• 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 1995年07月 [査読有り][通常論文]
   

  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 1995年04月 [査読有り][通常論文]
   

  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 1995年01月 [査読有り][通常論文]
   

  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 1994年08月 [査読有り][通常論文]
   

  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 1994年08月 [査読有り][通常論文]
   

  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 1994年06月 [査読有り][通常論文]
   

  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 1994年05月 [査読有り][通常論文]
   

  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 1993年12月 [査読有り][通常論文]
   

  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 1993年08月 [査読有り][通常論文]
   

  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.
• 低カゼイン食にメチオニンおよびその誘導体を補足したときのラット成長,肝脂質に及ぼす影響と吸収速度との関連

  葛西 隆則, 園山 慶, 桐山 修八

  日本農芸化学会誌 61 11 1425 - 1434 社団法人 日本農芸化学会 1987年11月 [査読有り][通常論文]
   

  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 1986年08月 [査読有り][通常論文]
  

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年07月
• 通常飼育マウス消化管における生存と定着に寄与するビフィズス菌遺伝子のINSeq法による同定

  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年
• 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年04月
• 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

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• 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年
• 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年04月
• 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

  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年
• 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年
• 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年
• 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年
• 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年04月
• 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年04月
• 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年
• 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年
• 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年
• 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年
• 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年
• 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年
• 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年
• 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年
• 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年
• 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年 [査読無し][招待有り]
  
• 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年 [査読無し][招待有り]
  
• 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年
• 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年
• 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年
• 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年 [査読無し][招待有り]
  
• 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年
• 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年
• 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年
• 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年
• 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年
• 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年
• 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

  園山 慶 The Lipid 27 172 -178 2016年 [査読無し][招待有り]
  
• 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年
• 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年
• 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年04月 [査読無し][通常論文]
  
• 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年
• 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年
• 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年 [査読無し][招待有り]
  
• 腸内微生物とアレルギー発症との関係および難消化性オリゴ糖によるその制御に関する研究

  園山 慶 ルミナコイド研究 16 (1) 1 -8 2012年 [査読無し][招待有り]
  
• 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年 [査読無し][招待有り]
  
• 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年10月 [査読無し][招待有り]
  
• 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年07月 [査読無し][招待有り]
  
• メタボリックシンドロームと腸内細菌叢

  園山 慶 腸内細菌学雑誌 = Journal of intestinal microbiology 24 (3) 193 -201 2010年07月 [査読無し][招待有り]
   

  肥満およびそれに関連した糖尿病などのメタボリックシンドロームの増加は世界的な問題となっている．最近の研究により，腸内細菌叢がこれらの疾患に関与する環境要因のひとつであることが明らかになってきた．無菌マウスを用いた研究は，腸内細菌が宿主における食餌からのエネルギー獲得および脂質・エネルギー代謝に影響することにより体脂肪蓄積に重要な役割を果たしていることを示唆した．また，肥満個体と正常体重個体との間で腸内細菌叢が異なることが，実験動物およびヒトにおいて観察された．さらに，腸内のグラム陰性細菌由来のリポ多糖が体内に移行して代謝性エンドトキシン血症を生じ，それが白色脂肪組織における軽度炎症，さらには全身性のインスリン抵抗性に寄与することが示された．これらの知見は腸内細菌叢が肥満およびメタボリックシンドロームの予防・治療の標的となりうることを示唆する．実際，プレバイオティクスおよびプロバイオティクスがこれらの疾患を予防・改善することが報告されている．本総説では，この数年で急速に進展した腸内細菌叢と肥満およびメタボリックシンドロームとの関係に関する研究を紹介する．
• 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年05月 [査読無し][招待有り]
  
• α-ガラクトシル基を含むオリゴ糖の機能と応用

  橋本 博之, 園山 慶 化学と生物 43 (7) 431 -438 2005年07月 [査読無し][招待有り]
  
• 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年
• 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年
• α-ガラクトシル基を含むオリゴ糖の合成とその機能

  橋本 博之, 山下 亜希子, 藤田 孝輝, 岡田 正通, 森 茂治, 園山 慶, 北畑 寿美雄 Journal of applied glycoscience 51 (2) 169 -176 2004年04月20日 [査読無し][通常論文]
  
• 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年
• 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年
• 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年
• 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年
• 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年11月01日 [査読無し][通常論文]
  
• 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年11月 [査読無し][招待有り]
  
• 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年
• 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年
• 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年08月25日 [査読無し][通常論文]
  
• 難治性の小麦アレルギーに挑む アレルゲンの同定から対応食品の開発まで

  田辺 創一, 渡辺 純, 園山 慶, 渡辺 道子 化学と生物 39 (7) 440 -447 2001年07月25日 [査読無し][通常論文]
  
• 消化管におけるアポリポタンパク質の発現調節に関する研究

  園山 慶 日本栄養・食糧学会誌 : Nippon eiyo shokuryo gakkaishi = Journal of Japanese Society of Nutrition and Food Science 53 (2) 65 -70 2000年04月 [査読無し][招待有り]
   

  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年06月 [査読無し][招待有り]
  
• 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年07月 [査読無し][招待有り]
  

書籍等出版物

• 花粉症等アレルギー疾患予防食品の開発

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

  (担当:共編者(共編著者)範囲: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年
• 腸内細菌・口腔内細菌と全身疾患

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

  (担当:分担執筆範囲:オリゴ糖)
  シーエムシー出版 2015年
• 食品機能性成分の吸収・代謝機構

  (担当:分担執筆範囲:オリゴ糖)
  シーエムシー出版 2013年
• Handbook of diet, nutrition and the skin

  (担当:分担執筆範囲:Fructo-oligosaccharides and skin inflammation)
  Wageningen Academic Publishers 2012年
• ルミナコイド研究のフロンティア

  (担当:分担執筆範囲: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年

講演・口頭発表等

• Exosome, a possible mediator of crosstalk between gut microbiota and host  [招待講演]

  In Symposium 37 Exosome & microRNA, 7th International Conference on Food Factors 2019年12月 口頭発表（招待・特別）
• 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  [招待講演]

  第104回日本栄養・食糧学会関東支部大会シンポジウム 2019年09月 口頭発表（招待・特別）
• 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  [招待講演]

  日本農芸化学会2019年度大会シンポジウム 2019年03月 口頭発表（招待・特別）
• 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  [招待講演]

  第75回日本栄養・食糧学会中部支部大会特別講演 2018年12月 口頭発表（招待・特別）
• 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  [招待講演]

  2018年度 日本乳酸菌学会泊まり込みセミナー 2018年05月 公開講演，セミナー，チュートリアル，講習，講義等
• 腸内細菌叢の健康機能を媒介する細胞外小胞  [招待講演]

  日本栄養・食糧学会第72回大会シンポジウム 2018年05月 口頭発表（招待・特別）
• 腸管オルガノイドを用いたmicroRNAの解析  [招待講演]

  日本農芸化学会2018年度大会シンポジウム 2018年03月 口頭発表（招待・特別）
• アレルギーの危険因子としての消化管常在真菌  [招待講演]

  日本アレルギー学会第66回学術大会シンポジウム 2017年06月 口頭発表（招待・特別）
• 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  [招待講演]

  日本栄養・食糧学会第70回大会シンポジウム 2016年05月 口頭発表（招待・特別）
• 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  [招待講演]

  日本農芸化学会2016年度大会シンポジウム 2016年03月 口頭発表（招待・特別）
• Communication tools mediating the health-promoting action of prebiotics and probiotics  [招待講演]

  In Symposium 11 Immune Regulation by Gut Microbiota, Probiotics, and Prebiotics, 12th Asian Congress of Nutrition 2015年05月 口頭発表（招待・特別）
• 肥満，食餌および腸内細菌叢の関係におけるAkkermansia muciniphilaの意義  [招待講演]

  第35回日本肥満学会 シンポジウム 2014年10月 口頭発表（招待・特別）
• メタボリック・シンドロームとプロバイオティックス  [招待講演]

  日本臨床栄養学会 特別セミナー in 札幌 2014年07月 公開講演，セミナー，チュートリアル，講習，講義等
• 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  [招待講演]

  日本栄養・食糧学会第68回大会シンポジウム 2014年06月 口頭発表（招待・特別）
• 腸内細菌叢とメタボリックシンドローム  [招待講演]

  日本細菌学会第86回総会シンポジウム 2013年03月 口頭発表（招待・特別）
• 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  [招待講演]

  日本食物繊維学会第16回学術集会シンポジウム 2011年11月 口頭発表（招待・特別）
• Prevention of murine contact hypersensitivity by prebiotics  [招待講演]

  International Symposium at the 7th Annual Meeting of Japanese Association for Food Immunology 2011年10月 口頭発表（招待・特別）
• Prevention of allergic disease by use of prebiotics – animal studies  [招待講演]

  International Symposium at Annual Meeting of Korean Society of Food Science and Technology 2010年06月 口頭発表（招待・特別）
• Prevention of allergic disease by prebiotics – animal studies  [招待講演]

  日本農芸化学会2010年度大会シンポジウム 2010年03月 口頭発表（招待・特別）
• 難消化性糖類のアレルギー抑制作用  [招待講演]

  第63回日本栄養・食糧学会大会シンポジウム 2009年05月 口頭発表（招待・特別）
• 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  [招待講演]

  日本食物繊維学会第13回学術集会シンポジウム 2008年11月 口頭発表（招待・特別）
• 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  [招待講演]

  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年11月 口頭発表（招待・特別）
• 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  [招待講演]

  日本栄養・食糧学会中部支部大会シンポジウム 2007年07月 口頭発表（招待・特別）
• Is gastrointestinal colonization by Candida albicans a risk factor for food allergy?  [招待講演]

  Second International Biomicrocosmos Workshop for the Enhancement of Gastro-Intestinal Biosphere Research 2006年02月 口頭発表（招待・特別）
• 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  [招待講演]

  日本食品免疫学会第1回学術大会シンポジウム 2005年11月 口頭発表（招待・特別）
• 食物繊維による腸管免疫の修飾  [招待講演]

  Hindgut Club 第３回セミナー 1997年12月 口頭発表（招待・特別）
• 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  [招待講演]

  第19回日本臨床栄養学会総会シンポジウム 1997年11月 口頭発表（招待・特別）

所属学協会

• 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   

共同研究・競争的資金等の研究課題

• 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

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  研究期間 : 2020年04月 -2023年03月 

  代表者 : 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。
• 食品-腸内細菌-宿主の連関を媒介する循環血中のmicroRNA

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  研究期間 : 2017年04月 -2020年03月 

  代表者 : 園山 慶

   

  本研究では、難消化性オリゴ糖摂取がマウスの結腸粘膜固有層リンパ球における一群のmicroRNA（miRNA）発現を増加させることを見出した。それらのmiRNAの多くは、通常マウスにおいて無菌マウスに比して高発現するmiRNAと一致した。これらの知見は腸内細菌が宿主の腸管免疫の恒常性に影響を及ぼすときにmiRNAによる遺伝子サイレンシングが関与することを示唆する。また、これらのmiRNAのmimicをヒトT細胞リンパ球細胞株Jurkatに導入し、標的遺伝子を探索するとともにT細胞分化への影響を解析する実験系を確立した。
• 腸内細菌叢存在下でのビフィズス菌の腸内活動に重要な遺伝子の同定と機能検証

  日本学術振興会：科学研究費助成事業 基盤研究(B)

  研究期間 : 2016年04月 -2019年03月 

  代表者 : 吹谷 智, 小椋 義俊, 園山 慶

   

  健康に有用な腸内細菌として知られるビフィズス菌が腸内でどのように活動しているのかについて，分子レベルでの機構を明らかにするために，まず通常飼育マウス腸内でヒト由来ビフィズス菌の長期定着が可能な飼育試験系を確立した．この系を用いて，腸内特異的に発現する遺伝子を薬剤耐性の変化により検出するR-IVET法による解析を行い，腸内特異的に発現する遺伝子を同定した．さらに，48,000株からなるビフィズス菌のトランスポゾン変異株集団を構築して無菌マウスに投与し，各変異株の腸内での増減をINSeq法により包括的に評価した結果，腸内での生存と定着に重要と考えられる442個の遺伝子を同定することができた．
• 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）

  研究期間 : 2014年04月 -2017年03月 

  代表者 : 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）

  研究期間 : 2013年04月 -2015年03月 

  代表者 : 園山 慶
• 体脂肪蓄積を制御するシンバイオティクス開発の基盤解析

  文部科学省：科学研究費補助金(基盤研究(B))

  研究期間 : 2011年 -2012年 

  代表者 : 園山 慶

   

  我々はこれまでに、マウスを用いて乳酸菌株Lactobacillus plantarum No.14の抗肥満効果を報告したが、腸内細菌が宿主の脂肪組織に影響をおよぼす細胞分子機序は不明である。本研究では、No.14株が2型糖尿病モデルマウスの肥満、脂肪組織炎症、およびインスリン抵抗性を抑制することを示すとともに、動物実験と培養細胞を用いて腸管から脂肪組織への情報伝達について解析し、細胞外小胞であるエキソソームが細胞間コミュニケーションを担う可能性を示唆した。
• 冬眠動物における腸内細菌叢と腸粘膜バリア:新しい共生様式の解明

  文部科学省：科学研究費補助金(挑戦的萌芽研究)

  研究期間 : 2011年 -2011年 

  代表者 : 園山 慶

   

  冬眠動物における腸内細菌の制御と腸粘膜バリアについて解析した。シリアンハムスターの冬眠個体では小腸管腔内の分泌抗体濃度が高く、感染防御に貢献している可能性が考えられた。実際、冬眠ハムスターでは腸間膜リンパ節への細菌の移行は認められなかった。また免疫組織におけるリンパ球プロファイルの解析の結果、冬眠期においても消化管粘膜における感染防御機能および免疫制御機能が維持されていると推察された。さらに、冬眠ハムスターの小腸粘膜上皮における細胞更新が著しく遅滞しており、それにより細胞寿命の延長にともなう細胞老化が生じていることが推察された。
• 食品による消化管微生物環境の制御を介した免疫疾患予防の基盤解析

  文部科学省：科学研究費補助金(基盤研究(B))

  研究期間 : 2007年 -2008年 

  代表者 : 園山 慶, 渡辺 純

   

  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)

  研究期間 : 2007年 -2008年 

  代表者 : 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.
• 経口免疫寛容誘導を介した食物アレルギー抑制機能を有する食品開発のための基盤解析

  文部科学省：科学研究費補助金(若手研究(B))

  研究期間 : 2006年 -2008年 

  代表者 : 渡辺 純, 園山 慶, 伊藤 進, 田口 秀典

   

  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。
• 食餌由来オリゴ糖による免疫応答・炎症反応制御の細胞・分子基盤

  文部科学省：科学研究費補助金(基盤研究(B))

  研究期間 : 2004年 -2006年 

  代表者 : 園山 慶, 福島 道広

   

  α-結合ガラクトオリゴ糖(α-GOS)の摂取がアレルギー反応を抑制する機構を明らかにするため、以下の検討を行った。1.α-GOSによるI型アレルギーの抑制に、α-ガラクトシル基をもつ糖脂質により活性化されることが知られているNKT細胞が関与すると想定したConA誘導肝炎およびエンドトキシンショックのマウスモデルを用いて検討した結果、α-GOSは抑制効果を示さなかった。また、マウスの脾細胞をex vivoで培養し、LPS、抗原、ならびにNKT細胞の内因性リガンドおよび外因性リガンドで刺激するとIFN-γ産生が増加し、また抗CD1d抗体の添加により抑制されたので、このときのIFN-γ産生にはCD1d拘束性のNKT細胞活性化が寄与することが示唆されたが、α-GOSを培地に添加してもIFN-γ産生は変化しなかった。以上より、NKT細胞活性化にα-GOSは影響しないことが示唆された。2.α-GOSは炎症性細胞浸潤を抑制するので、その機構について検討した。感作マウスの腹腔に抗原を投与した後の腹腔洗浄液に含まれる細胞を計数することにより、in vivoにおける細胞浸潤を評価した結果、α-GOS摂取により細胞浸潤が減少することが示された。また、腹腔浸潤細胞あるいは末梢血白血球はin vitroにおいて腹腔洗浄液上清に対して走化性を示し、これはα-GOS摂取マウスの腹腔洗浄液上清で低下した。し...
• 雑豆類・きのこの難消化性澱粉・食物繊維投与によるラットの脂質代謝とその機構解析

  文部科学省：科学研究費補助金(基盤研究(C))

  研究期間 : 2003年 -2004年 

  代表者 : 福島 道広, 園山 慶, 関川 三男, 小嶋 道之, 島田 謙一郎

   

  豆はその中に含まれている食物繊維、澱粉、ポリフェノールなどの成分が生理機能として有用であることが知られているが、その生理的機構については未だ明らかにされていない。また、きのこに含まれている食物繊維の健康機能性についても明らかにされていない。そこで本研究は、豆類としてアズキ、キントキ及びテボウのレジスタントスターチ及びきのこ食物繊維の脂質代謝に及ぼすメカニズムを明らかにした。その結果は以下の通りです。1.アズキ、キントキ及びテボウの皮からの食物繊維の機能性については血清コレステロール濃度に影響を及ぼさなかった。次にアズキ、キントキ及びテボウの餡を製造する過程で生成されるレジスタントスターチ及びその未消化物には血清コレステロール低下作用がみられ、LDL-receptor及びSR-B1の増加による肝臓へのLDL及びHDLの取り込みの増加及びCholesterol7α-hydroxylase活性の上昇による糞便中への胆汁酸類の排泄促進等に起因している可能性を明らかにした。2.アズキの水溶性(ポリフェノール)画分には,アセトアミノフェンのラット慢性肝毒性に対して,過酸化脂質の抑制及び抗酸化酵素(カタラーゼ,グルタチオン還元酵素)活性の増加がみられ,抗酸化活性能を有していることを明らかにした.3.きのこ食物繊維にも血清コレステロール低下作用が認められ、LDL-receptorの増加による...
• 食事成分による大腸ガン細胞の増殖抑制・細胞死誘導におけるアクチビンAの役割の解析

  文部科学省：科学研究費補助金(基盤研究(C))

  研究期間 : 2001年 -2002年 

  代表者 : 園山 慶

   

  1.食物繊維による大腸ガン抑制作用の一部は食物繊維の腸内発酵産物である酪酸により説明されている。本研究では、酪酸が結腸ガン細胞の増殖停止及びアポトーシスを誘導する際に増殖因子アクチビンAがメディエーターとなるか否かを明らかにすることを目的として、以下の検討を行った。2.ヒト結腸ガン細胞株HT-29を酪酸添加培地で培養し、細胞周期をフローサイトメーターで分析した結果、G1期停止及びアポトーシスが誘導されることが示された。また遺伝子発現を半定量的RT-PCRで分析した結果、アクチビンA mRNAレベルが酪酸により時間ならびに濃度依存的に著しく増加した。トリコスタチンAによっても発現が増加したことから、酪酸の影響はヒストンアセチラーゼの阻害によることが示唆された。しかしながら各種正常細胞株ならびにラット結腸組織断片では、アクチビンA mRNAレベルは酪酸により変化しなかったので、酪酸によるアクチビンA遺伝子発現の刺激はガン細胞に特異的な現象であると推察された。3.外因性のアクチビンAがHT-29細胞の増殖に及ぽす影響をBrdU取り込みならびにフローサイトメーターにより分析したが、変化は見られなかった。そこで、アクチビンAの細胞内情報伝達を担うsmadタンパクのリン酸化を免疫沈降及びウェスタン分析により調べた。アクチビンAはsmad2のリン酸化を誘導したが、smad2/3/4複合体の...
• アレルギーの予防・治療を意図した食品創製のための基盤解析

  文部科学省：科学研究費補助金(基盤研究(B))

  研究期間 : 2001年 -2002年 

  代表者 : 葛西 隆則, 渡辺 純, 園山 慶

   

  1.小麦粉中のアレルゲンを網羅的に酵素分解することにより調製した低アレルゲン化小麦粉ならびに難消化性オリゴ糖ラフィノースのアレルギー改善機構を明らかにすることを目的として、以下の検討を行った。2.Brown Norway(BN)ラットを用いてグルテン特異的アレルギー性気道好酸球性炎症モデルを確立し、低アレルゲン化小麦粉の気道炎症予防効果を明らかにした。また低アレルゲン化小麦粉によりグルテン特異的な全身性の免疫応答が抑制されたので、低アレルゲン化小麦粉は経口免疫寛容を誘導することによりアレルギーを予防することが示唆された。そこでBALB/cマウスを用いて免疫細胞養子移入実験を行ったところ、低アレルゲン化小麦粉を摂取したマウスをドナーとした場合にレシピエントのグルテン特異的血清抗体価が低く抑えられた。したがって低アレルゲン化小麦粉による経口免疫寛容誘導には抑制性T細胞が出現するアクティブサプレッションが関与することが示唆された。また、低アレルゲン化小麦粉は卵白アルブミン(OVA)の腸管透過を抑制するペプチドを含むことが明らかとなり、BNラットにおけるOVAの経口感作を低アレルゲン化小麦粉が抑制することが示された。3.OVAを抗原としたBNラットの気道好酸球性炎症モデルにおいて、食餌ラフィノースは気道炎症を改善し、この効果は盲腸を切除してネオマイシンを毎日経口投与した動物においても...
• 小腸上皮細胞におけるアポリポタンパクA-IV遺伝子発現の神経内分泌制御の解析

  文部科学省：科学研究費補助金(基盤研究(C))

  研究期間 : 1999年 -2000年 

  代表者 : 園山 慶

   

  筆者らのこれまでの研究により、小腸におけるアポリポタンパク(アポ)A-IV遺伝子の発現が管腔因子(胆汁成分)及び体液性因子(消化管ホルモンPYY)による調節を受けることが示唆されている。本研究では、アポA-IV遺伝子の発現調節に関わる神経性因子について検討するとともに、内分泌因子について更に広範に検索した。各種の神経伝達ブロッカーをラットに投与する実験を行い、神経節遮断薬、コリン作動性神経拮抗薬、及びNO合成酵素阻害薬が回腸アポA-IV mRNAレベルを低下させたことから、回腸におけるアポA-IV遺伝子の基礎的な発現は少なくとも一個の神経節を有するコリン作動性神経及びNO作動性神経の支配を受けることが示唆された。また、β-アドレナリン作動性神経拮抗薬、ドパミン作動性神経拮抗薬、セロトニン作動性神経拮抗薬、アデノシン作動性神経拮抗薬、及びATP作動性神経拮抗薬は影響を及ぼさなかった。更に、食餌脂質や小腸切除後の適応における回腸アポA-IV遺伝子発現の増加に神経節遮断薬が影響を及ぼさなかったことから、これらに神経系は関与しないと考えられた。小腸上皮細胞のモデル系としてヒト結腸ガン由来細胞株Caco-2を用いて、さまざまな消化管ホルモンならびに神経ペプチドがアポA-IV遺伝子の発現に及ぼす影響を解析した結果、PYYは濃度及び時間依存的にアポA-IV mRNAレベルを増加させ、またV...
• 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))

  研究期間 : 1997年 -1998年 

  代表者 : 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...
• 消化管機能の生理・生化学的解析

  文部科学省：科学研究費補助金(基盤研究(A))

  研究期間 : 1997年 -1998年 

  代表者 : 青山 頼孝, 石塚 敏, 園山 慶, 川端 潤, 葛西 隆則

   

  1. オロト酸の摂取によって肝脂質の増加はカゼインをタンパク質源として用いると全卵タンパク質、大豆タンパク質、小麦グルテンをタンパク質源とするときに比較して著しく顕著であることを見出した。カゼインをタンパク質源とする食餌にオロト酸を添加すると肝脂質が著しく蓄積し、肝スーパーオキシドジスムターゼ活性は低下した。事実ラットの血清オルニチンカルバモイルトランスフェラーゼ活性は有意に増加した。このことは肝障害をひきおこしていることが推定された。さらに肝1,2-ジアシルグリセロールレベルはオロト酸摂取時において約10倍に増加した。以上の結果、オロト酸摂取時はセカンドメッセンジャーを増加させ、肝スーパーオキシドの増加が肝障害を引き起こしている可能性が示唆された。2. アセチルCoAカルボキシラーゼ阻害物質としてトウガラシよりE、E-およびE-9 -octadeca-10,12-dienoic asidを見出した。この阻害活性は脂肪数より約50倍高かった。3. 大腸ガン活発剤である1,2-ジメチルヒドラジン(DMH)誘発前ガン病変はテンサイ食物繊維の摂取によって抑制することをすでに見出した。γ線による前ガン病変に対してもテンサイ食物繊維は影響はないが、抗アシアロ GM1投与下では前ガン病変の増加を有意に抑制した。4. DMH投与後、結腸上皮にアポトーシス、p53の誘導を観察した。さらに、紫...
• 食物繊維の大腸ガン抑制効果における大腸上皮細胞アポトーシスの役割

  文部科学省：科学研究費補助金(基盤研究(C))

  研究期間 : 1997年 -1998年 

  代表者 : 葛西 隆則, 石塚 敏, 園山 慶

   

  食物繊維がもつ大腸ガン抑制効果の分子機構について、遺伝子損傷ストレス後のアポトーシスに焦点をあて解析した。ラットに対する化学発ガン剤ジメチルヒドラジン(DMH)の投与により結腸上皮細胞にアポトーシスが誘導され、ビート食物繊維がその頻度を増加させたことから、遺伝子損傷細胞のアポトーシスによる除去効率の上昇が食物繊維の大腸ガン抑制作用に関連することが示唆された。ラット結腸におけるDMH投与による遺伝子損傷ストレス後の遺伝子発現を半定量的RT-PCRにより解析したところ、組織学的にアポトーシスが検出されるのに先立って、ガン抑制遺伝子産物p53の標的遺伝子であるサイクリン依存性キナーゼインヒビターp21及びアポトーシス誘発タンパクBaxの遺伝子発現が増加することが確認され、更に、TGF-βスーパーファミリーの一員であるアクチビンのβAサブユニット遺伝子の発現も増加することが新たに見出された。このような現象をより詳細に解析可能なin vitroモデルを確立するためにラット小腸上皮由来正常細胞株IEC-6に遺伝子損傷ストレスとして紫外線を照射し、その後の細胞生存率と遺伝子発現を解析した。その結果、紫外線照射により時間及び照射エネルギー量依存的に生存率の低下が認められ、それはアポトーシスによることがDNAのアガロースゲル電気泳動により示された。また、ラット結腸で観察されたと同様にアクチビン...
• 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))

  研究期間 : 1996年 -1996年 

  代表者 : 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))

  研究期間 : 1995年 -1995年 

  代表者 : 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...