In vivo bioluminescence imaging revealed the change of the time window of BDNF expression in the brain elicited by a single bout of exercise following repeated exercise
Ryo Ikegami, Takahiro Inoue, Yasuyuki Takamatsu, Taichi Nishio, Mamoru Fukuchi, Sanae Haga, Michitaka Ozaki, Hiroshi Maejima
Neuroscience Letters, 137830, 137830, Elsevier BV, 2024年05月
研究論文（学術雑誌）

Temporal dynamics of brain BDNF expression following a single bout of exercise: A bioluminescence imaging study.
Takahiro Inoue, Ryo Ikegami, Yasuyuki Takamatsu, Mamoru Fukuchi, Sanae Haga, Michitaka Ozaki, Hiroshi Maejima
Neuroscience letters, 799, 137120, 137120, 2023年03月16日, ［国際誌］
英語, 研究論文（学術雑誌）, Physical exercise increases brain-derived neurotrophic factor (BDNF) expression in the brain. However, the absence of non-invasive and repetitive monitoring of BDNF expression in the brains of living animals has limited the understanding of how BDNF expression changes after exercise. This study aimed to elucidate the temporal dynamics of BDNF expression in the brain after a single bout of exercise, using in vivo bioluminescence imaging. This study included Bdnf-Luc mice with a firefly Luciferase gene inserted at the translation start site of the mouse Bdnf gene. BDNF expression was evaluated based on the luminescence signal of the luciferase substrate administered to mice. Bioluminescence imaging was performed at 0, 1, 3, 6, 12, and 24 h after treadmill exercise (15 m/min for 1 h). Compared to the sedentary condition of each mouse, the luminescence signal increased by approximately 60 % between 1 and 3 h after exercise. The luminescence signal remained slightly increased by approximately 20 % even 6-24 h after exercise. This study is the first to demonstrate exercise-enhanced BDNF expression in the brains of living animals. These results provide evidence that a single bout of exercise transiently increases BDNF expression in the brain within a limited time window.

The usefulness of measuring n-butyric acid concentration as a new indicator of blood decomposition in forensic autopsy.
Kotaro Matoba, Manabu Murakami, Emi Fujita, Shigeki Jin, Ryosuke Ogasawara, Tomoko Matoba, Akiko Takeuchi, Sanae Haga, Michitaka Ozaki, Hideki Hyodoh
Legal medicine (Tokyo, Japan), 57, 102071, 102071, 2022年04月15日, ［国際誌］
英語, 研究論文（学術雑誌）, In forensic medicine, although various alcohols have been reported as indicators of decomposition in collected blood, no studies have examined short-chain fatty acids as indicators. In this study, the blood n-butyric acid concentration was quantified, and the association between n-butyric acid and decomposition was investigated to determine whether the detection of n-butyric acid could be a new indicator of decomposition. Among the forensic autopsies performed from 2016 to 2018 in our laboratory, the cases were divided into decomposed (n = 20) and non-decomposed (n = 20) groups based on macroscopic findings. Blood samples collected at the time of autopsy were derivatized with 3-nitrophenylhydrazine hydrochloride after solid-phase extraction. The n-butyric acid concentration was measured using liquid chromatography-tandem mass spectrometry. In addition, ethanol and n-propanol were measured using a gas chromatography-flame ionization detector. There was a significant difference (p < 0.01) in the concentrations of n-butyric acid between the decomposed and non-decomposed groups (0.343 ± 0.259 [0.030-0.973] and 0.003 ± 0.002 [0.001-0.007] mg/mL, respectively). In the decomposed group, n-butyric acid was detected at high concentrations, even in cases where n-propanol was low. These results suggest that n-butyric acid is more likely to be an indicator of blood decomposition than n-propanol.

Poly(ADP-ribose) polymerase (PARP) is critically involved in liver ischemia/reperfusion-injury
S. Haga, A. Kanno, N. Morita, S. Jin, K. Matoba, T. Ozawa, M. Ozaki
The Journal of Surgical Research, 270, 124, 138, 2021年10月, ［査読有り］
英語, 研究論文（学術雑誌）

Period1 gene expression in the olfactory bulb and liver of freely moving streptozotocin-treated diabetic mouse.
Harumi Kanou, Kouki Nagasawa, Yuki Ishii, Aya Chishima, Juri Hayashi, Sanae Haga, Kenneth Sutherland, Masayori Ishikawa, Michitaka Ozaki, Hiroki Shirato, Kazuko Hamada, Toshiyuki Hamada
Biochemical and biophysical research communications, 560, 14, 20, 2021年06月30日, ［国際誌］
英語, 研究論文（学術雑誌）, Clock genes express circadian rhythms in most organs. These rhythms are organized throughout the whole body, regulated by the suprachiasmatic nucleus (SCN) in the brain. Disturbance of these clock gene expression rhythms is a risk factor for diseases such as obesity. In the present study, to explore the role of clock genes in developing diabetes, we examined the effect of streptozotocin (STZ)-induced high glucose on Period1 (Per1) gene expression rhythm in the liver and the olfactory bub (OB) in the brain. We found a drastic increase of Per1 expression in both tissues after STZ injection while blood glucose content was low. After a rapid expression peak, Per1 expression showed no rhythm. Associated with an increase of glucose content, behavior became arrhythmic. Finally, we succeeded in detecting an increase of Per1 expression in mice hair follicles on day 1 after STZ administration, before the onset of symptoms. These results show that elevated Per1 expression by STZ plays an important role in the aggravation of diabetes.

Sample preparation method with ultrafiltration for whole blood thiosulfate measurement.
Shigeki Jin, Manabu Murakami, Kotaro Matoba, Tomoko Matoba, Sanae Haga, Michitaka Ozaki, Akiko Takeuchi, Hideki Hyodoh
Legal medicine (Tokyo, Japan), 47, 101765, 101765, 2020年07月23日, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, Quantitative analysis of thiosulfate is useful for diagnosing hydrogen sulfide poisoning. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) enables more rapid and sensitive measurements than previous methodologies. As simple measurements of blood thiosulfate concentration are affected by the blood matrix, blood is used as the solvent to prepare the standard solution for calibration curve generation. Thus, a large amount of blood devoid of thiosulfate is required. We developed a preparation method by incorporating an ultrafiltration step to overcome this limitation and generate a calibration curve using a standard solution prepared with pure water. We used this improved method to investigate the stability of thiosulfate in refrigerated samples. To compare the effects of refrigeration, blood samples were prepared using the following two methods: one sample was treated with a 50-kDa exclusion ultrafiltration membrane and the other was not treated. The samples were stored at 4 °C, and then measured at 0, 3, 6, 24, 48, and 96 h. The incorporation of the ultrafiltration step in the measurement procedure enabled the quantification of thiosulfate, by plotting a calibration curve using a standard of pure water; it did not require a blood standard. Additionally, the reduction in whole blood thiosulfate concentration was within 10% during 2 days of refrigeration. Thus, the need for a large amount of blood to prepare the standard solution was resolved by the ultrafiltration step in test sample preparation. This method is useful to measure thiosulfate concentration and is not hindered by sample refrigeration for a few days.

Extracts of bilberry (Vaccinium myrtillus L.) fruits improve liver steatosis and injury in mice by preventing lipid accumulation and cell death.
Sanae Haga, Yimin, Hikari Yamaki, Shigeki Jin, Tetsuya Sogon, Naoki Morita, Michitaka Ozaki
Bioscience, Biotechnology, and Biochemistry, 83, 11, 2110, 2120, Oxford University Press (OUP), 2019年06月, ［査読有り］
英語, 研究論文（学術雑誌）,
Bilberry has been reported to have anti-oxidant and anti-inflammatory properties. We studied the effect of bilberry (Vaccinium myrtillus L.) fruits extracts (BEs) on the pathogenesis caused by lipid accumulation in fatty liver and non-alcoholic steatohepatitis (NASH). 5 μg/ml of BEs was enough to suppress lipid accumulation in the fatty liver model of the mouse hepatic AML12 cells. BEs increased cell viability and anti-oxidant capacity, presumably by activating (phosphorylating) Akt/STAT3 and inducing MnSOD/catalase. BEs also significantly reduced Rubicon and induced p62/SQSTM1, possibly contributing to reduce cellular lipids (lipophagy). When the mice were fed supplemented with BEs (5% or 10%, w/w), hepatic steatosis, injury, and hypercholesterolemia/hyperglycemia were significantly improved. Furthermore, histological and cytokine studies indicated that BEs possibly suppress hepatic inflammation (hepatitis) and fibrosis. Therefore, BEs improved liver steatosis and injury, and potentially suppress fibrosis by suppressing inflammatory response, which therefore may prevent the progression of fatty liver to NASH.

Activation of caspase-3 during Chlamydia trachomatis-induced apoptosis at a late stage.
Junji Matsuo, Sanae Haga, Kent Hashimoto, Torahiko Okubo, Takeaki Ozawa, Michitaka Ozaki, Hiroyuki Yamaguchi
Canadian Journal of Microbiology, 65, 2, 135, 143, 2019年02月, ［査読有り］, ［国際誌］
英語, 研究論文（学術雑誌）, The obligate intracellular bacterium Chlamydia trachomatis activates the host cell apoptosis pathway at a late stage of its developmental cycle. However, whether caspase-3, which is a key enzyme of apoptosis, is activated in Chlamydia-infected cells remains unknown. Here, we established HEp-2 cells stably expressing cFluc-DEVD, which is a caspase-3 substrate sequence inserted into cyclic firefly luciferase, and then monitored the dynamics of caspase-3 activity in cells infected with Chlamydia. Transfected cells without infection showed a significant increase in luciferase activity due to stimulation with staurosporine, an inducer of apoptosis. Activation was significantly blocked by addition of caspase inhibitor z-VAD-fmk. Furthermore, as expected, Chlamydia infection caused a significant increase in luciferase activation at 36-48 h postinfection with a contrastive decrease at 24 h postinfection, which is already well known. Such activation caused by the infection was much stronger when the amount of bacteria was increased. Thus, caspase-3 activation was accurately monitored by the luciferase activity in HEp-2 cells constitutively expressing the cFluc-DEVD probe. Furthermore, our data showed that C. trachomatis activates caspase-3 in host cells at a late stage of infection.

Detection of necroptosis in ligand-mediated and hypoxia-induced injury of hepatocytes using a novel optic probe-detecting receptor-interacting protein (RIP)1/RIP3 binding
Sanae Haga, Akira Kanno, Takeaki Ozawa, Naoki Morita, Mami Asano, Michitaka Ozaki
Oncology Research, 26, 3, 503, 513, Cognizant Communication Corporation, 2018年, ［査読有り］
英語, 研究論文（学術雑誌）, Liver injury is often observed in various pathological conditions including posthepatectomy state and cancer chemotherapy. It occurs mainly as a consequence of the combined necrotic and apoptotic types of cell death. In order to study liver/hepatocyte injury by the necrotic type of cell death, we studied signal-regulated necrosis (necroptosis) by developing a new optic probe for detecting receptor-interacting protein kinase 1 (RIP)/RIP3 binding, an essential process for necroptosis induction. In the mouse hepatocyte cell line, TIB-73 cells, TNF-a/cycloheximide (T/C) induced RIP1/3 binding only when caspase activity was suppressed by the caspase-specific inhibitor z-VAD-fmk (zVAD). T/C/zVAD-induced RIP1/3 binding was inhibited by necrostatin-1 (Nec-1), an allosteric inhibitor of RIP1. The reduced cell survival by T/C/zVAD was improved by Nec-1. These facts indicate that T/C induces necroptosis of hepatocytes when the apoptotic pathway is inhibited/unavailable. FasL also induced cell death, which was only partially inhibited by zVAD, indicating the possible involvement of necroptosis rather than apoptosis. FasL activated caspase 3 and, similarly, induced RIP1/3 binding when the caspases were inactivated. Interestingly, FasL-induced RIP1/3 binding was significantly suppressed by the antioxidants Trolox and N-acetyl cysteine (NAC), suggesting the involvement of reactive oxygen species (ROS) in FasL-induced necroptotic cellular processes. H2O2, by itself, induced RIP1/3 binding that was suppressed by Nec-1, but not by zVAD. Hypoxia induced RIP1/3 binding after reoxygenation, which was suppressed by Nec-1 or by the antioxidants. Cell death induced by hypoxia/ reoxygenation (H/R) was also improved by Nec-1. Similar to H2O2, H/R did not require caspase inhibition for RIP1/3 binding, suggesting the involvement of a caspase-independent mechanism for non-ligand-induced and/or redox-mediated necroptosis. These data indicate that ROS can induce necroptosis and mediate the FasL- and hypoxia-induced necroptosis via a molecular mechanism that differs from a conventional caspase-dependent pathway. In conclusion, necroptosis is potentially involved in liver/hepatocyte injury induced by oxidative stress and FasL in the absence of apoptosis.

Photo-activatable akt probe: A new tool to study the akt-dependent physiopathology of cancer cells
Sanae Haga, Takeaki Ozawa, Naoki Morita, Mami Asano, Shigeki Jin, Yimin, Michitaka Ozaki
Oncology Research, 26, 3, 467, 472, Cognizant Communication Corporation, 2018年, ［査読有り］
英語, 研究論文（学術雑誌）, Akt is commonly overexpressed and activated in cancer cells and plays a pivotal role in cell survival, protection, and chemoresistance. Therefore, Akt is one of the target molecules in understanding characters of cancer cells and developing anticancer drugs. Here we examined whether a newly developed photo-activatable Akt (PA-Akt) probe, based on a light-inducible protein interaction module of plant cryptochrome2 (CRY2) and cryptochrome-interacting basic helix–loop–helix (CIB1), can regulate Akt-associated cell functions. By illuminating blue light to the cells stably transfected with PA-Akt probe, CRY2-Akt (a fusion protein of CRY2 and Akt) underwent a structural change and interacted with Myr-CIBN (myristoylated N-terminal portion of CIB1), anchoring it at the cell membrane. Western blot analysis revealed that S473 and T308 of the Akt of probe-Akt were sequentially phosphorylated by intermittent and continuous light illumination. Endogenous Akt and GSK-3b, one of the main downstream signals of Akt, were also phosphorylated, depending on light intensity. These facts indicate that photo-activation of probe-Akt can activate endogenous Akt and its downstream signals. The photo-activated Akt conferred protection against nutritional deprivation and H2O2 stresses to the cells significantly. Using the newly developed PA-Akt probe, endogenous Akt was activated easily, transiently, and repeatedly. This probe will be a unique tool in studying Akt-associated specific cellular functions in cancer cells and developing anticancer drugs.

Evaluation and regulation cellular/organ functions by optic technology.
Michitaka Ozaki, Sanae Haga, Takeaki Ozawa, Naoki Morita, Toshiyuki Hamada
Organ Biology, 24, 2, 87, 91, 一般社団法人 日本臓器保存生物医学会, 2017年, ［査読有り］
英語, 研究論文（学術雑誌）, To develop an effective organ/cell preservation method and to monitor post-transplant graft function continuously and non-invasively, an innovative optic technology to visualize cell/organ function seems to be useful. We have developed some optic probes to visualize regulated cell death (apoptosis, necroptosis, pyroptosis), redox states and cellular stresses, pH and cellular antigens in deeper lesions of the organ. In the hepatic ischemia/reperfusion model of mice, we successfully imaged liver oxidative stress (by redox-sensitive GFP) and apoptosis (by caspase-3 activity) non-invasively and chronologically in a single mouse. We also developed a unique tool to visualize intracellular pH and succeeded in imaging dynamic changes of pH in a mouse posterior limb ischemia/reperfusion model. We are also developing the new devices for tissue/organ imaging. We are trying to monitor the optic signals chronologically and track the lesions in the body by developing light sensor and multiple CCD camera system, which can be used in endoscopic examination and surgical operation. It is still on a way, but this kind of technology will definitely provide a new avenue toward effective and non-invasive surgical therapy in the future.

Relevance of FXR-p62/SQSTM1 pathway for survival and protection of mouse hepatocytes and liver, especially with steatosis
Sanae Haga, Yimin, Michitaka Ozaki
BMC GASTROENTEROLOGY, 17, 1, 9, BIOMED CENTRAL LTD, 2017年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Background: Liver injury and regeneration involve complicated processes and are affected by various physiopathological conditions. Surgically, severe liver injury after surgical resection often leads to fatal liver failure, especially with some underlying pathological conditions such as steatosis. Therefore, protection from the injury of hepatocytes and liver is a serious concern in various clinical settings.
Methods: We studied the effects of the farnesoid X receptor (FXR) on cell survival and steatosis in mouse hepatocytes (AML12 mouse liver cells) and investigated their molecular mechanisms. We next studied whether or not FXR improves liver injury, regeneration and steatosis in a mouse model of partial hepatectomy (PH) with steatosis.
Results: An FXR-specific agonist, GW4064, induced expressions of the p62/SQSTM1 gene and protein in AML12 mouse liver cells. Because we previously reported p62/SQSTM1 as a key molecule for antioxidation and cell survival in hepatocytes, we next examined the activation of nuclear factor erythroid 2-related factor-2 (Nrf2) and induction of the antioxidant molecules by GW4064. GW4064 activated Nrf2 and subsequently induced antioxidant molecules (Nrf2, catalase, HO-1, and thioredoxin). We also examined expressions of pro-survival and cell protective molecules associated with p62/SQSTM1. Expectedly, GW4064 induced phosphorylation of Akt, expression of the anti-apoptotic molecules (Bcl-xL and Bcl-2), and reduced harmful hepatic molecules (Fas ligand and Fas). GW4064 promoted hepatocyte survival, which was cancelled by p62/SQSTM1 siRNA. These findings suggest the potential relevance of the FXR-p62/SQSTM1 pathway for the survival and protection of hepatocytes. Furthermore, GW4064 induced the expression of small heterodimer partners (SHP) and suppressed liver X receptor (LXR)-induced steatosis in hepatocytes, expecting the in vivo protective effect of FXR on liver injury especially with steatosis. In the hepatectomy model of db/db mice with fatty liver, pre-treatment by GW4064 significantly reduced post-PH liver injury (serum levels of LDH, AST & ALT and histological study) and improved steatosis. The key molecules, p62/SQSTM1, Nrf2 and SHP were upregulated in fatty liver tissue by GW4064 treatment.
Conclusions: The present study is the first to demonstrate the relevance of FXR-p62/SQSTM1 and-SHP in the protection against injury of hepatocytes and post-PH liver, especially with steatosis.

Inflammatory responses increase secretion of MD-1 protein
Richard Thomas Jennings, Erdenezaya Odkhuu, Akina Nakashima, Naoko Morita, Toshihiko Kobayashi, Ikuko Yamai, Miyako Tanaka, Takayoshi Suganami, Sanae Haga, Michitaka Ozaki, Yasuharu Watanabe, Yoshinori Nagai, Kiyoshi Takatsu, Takane Kikuchi-Ueda, Isao Ichimonji, Yoshihiro Ogawa, Hidekazu Takagi, Tatsuya Yamazaki, Kensuke Miyake, Sachiko Akashi-Takamura
INTERNATIONAL IMMUNOLOGY, 28, 10, 503, 512, OXFORD UNIV PRESS, 2016年10月, ［査読有り］
英語, 研究論文（学術雑誌）, Novel antibodies detect the increased level of soluble MD-1 during inflammation.Radioprotective 105 (RP105) is a type I transmembrane protein, which associates with a glycoprotein, MD-1. Monoclonal antibody (mAb)-mediated ligation of RP105/MD-1 robustly activates B cells. RP105/MD-1 is structurally similar to Toll-like receptor 4 (TLR4)/MD-2. B-cell responses to TLR2 and TLR4/MD-2 ligands are impaired in the absence of RP105 or MD-1. In addition to RP105/MD-1, MD-1 alone is secreted. The structure of MD-1 shows that MD-1 has a hydrophobic cavity that directly binds to phospholipids. Little is known, however, about a ligand for MD-1 and the role of MD-1 in vivo. To study the role of RP105/MD-1 and MD-1 alone, specific mAbs against MD-1 are needed. Here, we report the establishment and characterization of two anti-MD-1 mAbs (JR2G9, JR7G1). JR2G9 detects soluble MD-1, whereas JR7G1 binds both soluble MD-1 and the cell surface RP105/MD-1 complex. With these mAbs, soluble MD-1 was detected in the serum and urine. The MD-1 concentration was altered by infection, diet and reperfusion injury. Serum MD-1 was rapidly elevated by TLR ligand injection in mice. The quantitative PCR and supernatant-precipitated data indicate that macrophages are one of the sources of serum soluble MD-1. These results suggest that soluble MD-1 is a valuable biomarker for inflammatory diseases.

Development for the measurement of serum thiosulfate using LC-MS/MS in forensic diagnosis of H2S poisoning
Shigeki Jin, Hideki Hyodoh, Kotaro Matoba, Fei Feng, Akira Hayakawa, Katsuhiro Okuda, Keiko Shimizu, Sanae Haga, Michitaka Ozaki, Koichi Terazawa
LEGAL MEDICINE, 22, 18, 22, ELSEVIER IRELAND LTD, 2016年09月, ［査読有り］
英語, 研究論文（学術雑誌）, Thiosulfate measurement is crucial to diagnosis of hydrogen sulfide (H2S) poisoning in forensic toxicology. Although GC-MS method is currently regarded as a standard thiosulfate measurement, it requires complicated sample preparation prior to analysis. This study presents a simple, rapid, and highly sensitive method for the quantitative analysis of serum thiosulfate by using liquid chromatography-tandem mass spectrometry (LC-MS/MS). This method is based on selected reaction monitoring and has high sensitivity with a lower quantification limit of 0.5 mu M. Precision and accuracy of this method meet the basic requirements for quantitative analysis (intra- and inter-day tests have a relative standard deviation of <= 10.4%;range of analytical recovery is 94.3-102.6%). On the measurements of serum thiosulfate by our developed method, a thiosulfate concentration as 57.5 mu M was detected clearly in the H2S poisoning case comparing to the non poisoning case in which only a trace amount of thiosulfate was observed. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

Long-term ex vivo and in vivo monitoring of tumor progression by using dual luciferases
Naoki Morita, Sanae Haga, Yoshihiro Ohmiya, Michitaka Ozaki
ANALYTICAL BIOCHEMISTRY, 497, 24, 26, ACADEMIC PRESS INC ELSEVIER SCIENCE, 2016年03月, ［査読有り］
英語, 研究論文（学術雑誌）, We propose a new concept of tumor progression monitoring using dual luciferases in living animals to reduce stress for small animals and the cost of luciferin. The secreted Cypridina luciferase (CLuc) was used as an ex vivo indicator to continuously monitor tumor progression. On the other hand, the non-secreted firefly luciferase was used as an in vivo indicator to analyze the spatial distribution of the tumor at suitable time points indicated by CLuc. Thus, the new monitoring systems that use dual luciferases are available, allowing long-term bioluminescence imaging under minimal stress for the experimental animals. (C) 2015 Elsevier Inc. All rights reserved.

p62/SQSTM1を基軸とした新たな肝臓・肝細胞保護・機能維持メカニズム
尾崎 倫孝, 芳賀 早苗
Organ Biology, 23, 2, 168, 172, 一般社団法人 日本臓器保存生物医学会, 2016年
日本語, We have been investigating the mechanism of liver regeneration and injury for liver surgery and transplantation. We firstly studied, in normal liver, the central role of IL-6/Jak/STAT3 in post-PH mitotic response and protective mechanism from hepatocyte injury, and the essential role of PDK1/Akt in post-PH cell growth in case where mitotic response is impaired. In fatty liver, reduced expression of p62/SQSTM1 plays a crucial role in post-hepatectomy acute injury and delayed regeneration with enhanced oxidative stress- and death signal-mediated injury. Reduced expression of p62/SQSTM1 lead to FasL/Fas overexpression and suppressed antioxidant genes through Nrf-2 inactivation, which along with the hypo-responsiveness of Akt, caused post-hepatectomy necrotic/apoptotic liver injury and delayed regeneration. Our study importantly suggests the potential role of p62/SQSTM1 in protecting the liver from all kinds of injury, providing pro-survival property to hepatocytes/liver. Here, we report the pivotal roles of p62/SQSTM1 as a key player for cell/organ preservation, and the potential application for liver preservation and transplantation.

Mitochondrial delivery of Coenzyme Q(10) via systemic administration using a MITO-Porter prevents ischemia/reperfusion injury in the mouse liver
Yuma Yamada, Kohei Nakamura, Jiro Abe, Mamoru Hyodo, Sanae Haga, Michitaka Ozaki, Hideyoshi Harashima
JOURNAL OF CONTROLLED RELEASE, 213, 86, 95, ELSEVIER SCIENCE BV, 2015年09月, ［査読有り］
英語, 研究論文（学術雑誌）, We herein report on a mitochondrial therapeutic effect based on the delivery of coenzyme Q(10) (CoQ(10)), an anti-oxidant, to in vivo mitochondria using a MITO-Porter, a liposome-based mitochondrial delivery system that functions via membrane fusion. To evaluate the effects, we used a mouse liver ischemia/reperfusion injury (I/R injury) model, in which mitochondrial reactive oxygen species are overexpressed. We packaged CoQ(10) in the lipid phase of a MITO-Porter and optimized the mitochondrial fusogenic activities to produce the CoQ(10)-MITO-Porter. A histological observation of the carriers in the liver by confocal laser scanning microscopy was done and the accumulation of the carrier labeled with a radio isotope in the liver confirmed that the CoQ(10)-MITO-Porter was delivered to liver mitochondria via systemic injection. These analytical results permitted us to optimize the compositions of the CoQ(10)-MITO-Porter so as to permit it to efficiently accumulate in mouse liver mitochondria. Finally, we applied the optimized CoQ(10) -MITO-Porter to mice via tail vein injection, and hepatic I/R injury was then induced, followed by measuring serum alanine aminotransferase (ALT) levels, a marker of liver injury. We confirmed that the use of the CoQ(10)-MITO-Porter resulted in a significant decrease in serum ALT levels, indicating that in vivo mitochondrial delivery of the CoQ(10) via MITO-Porter prevents I/R injury in mice livers. This provides a demonstration of the potential use of such a delivery system in mitochondrial therapies. (C) 2015 Elsevier B.V. All rights reserved.

虚血再灌流臓器障害に関する基礎研究の進歩 基礎研究・移植再生 光イメージングによる虚血再灌流障害の動的・質的解析　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 野田 なつみ, 小澤 岳昌, 森田 直樹
日本外科学会定期学術集会抄録集, 115回, SY, 6, (一社)日本外科学会, 2015年04月
日本語

Growth Arrest and DNA Damage-Inducible 34 Regulates Liver Regeneration in Hepatic Steatosis in Mice
Yuka Inaba, Tomoko Furutani, Kumi Kimura, Hitoshi Watanabe, Sanae Haga, Yoshiaki Kido, Michihiro Matsumoto, Yasuhiko Yamamoto, Kenichi Harada, Shuichi Kaneko, Seiichi Oyadomari, Michitaka Ozaki, Masato Kasuga, Hiroshi Inoue
HEPATOLOGY, 61, 4, 1343, 1356, WILEY-BLACKWELL, 2015年04月, ［査読有り］
英語, 研究論文（学術雑誌）, The liver has robust regenerative potential in response to damage, but hepatic steatosis (HS) weakens this potential. We found that the enhanced integrated stress response (ISR) mediated by phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF2) impairs regeneration in HS and that growth arrest and DNA damage-inducible 34 (Gadd34)-dependent suppression of ISR plays a crucial role in fatty liver regeneration. Although mice fed a high-fat diet for 2 weeks developed moderate fatty liver with no increase in eIF2 phosphorylation before 70% hepatectomy, they showed impaired liver regeneration as a result of reduced proliferation and increased death of hepatocytes with increased phosphorylation of eIF2 and ISR. An increased ISR through Gadd34 knockdown induced C/EBP homologous protein (CHOP)-dependent apoptosis and receptor-interacting protein kinase 3-dependent necrosis, resulting in increased hepatocyte death during fatty liver regeneration. Furthermore, Gadd34 knockdown and increased phosphorylation of eIF2 decreased cyclin D1 protein and reduced hepatocyte proliferation. In contrast, enhancement of Gadd34 suppressed phosphorylation of eIF2 and reduced CHOP expression and hepatocyte apoptosis without affecting hepatocyte proliferation, clearly improving fatty liver regeneration. In more severe fatty liver of leptin receptor-deficient db/db mice, forced expression of hepatic Gadd34 also promoted hepatic regeneration after hepatectomy. Conclusion: Gadd34-mediated regulation of ISR acts as a physiological defense mechanism against impaired liver regeneration resulting from steatosis and is thus a possible therapeutic target for impaired regeneration in HS. (Hepatology 2015;61:1343-1356)

p62/SQSTM1 Plays a Protective Role in Oxidative Injury of Steatotic Liver in a Mouse Hepatectomy Model
Sanae Haga, Takeaki Ozawa, Yuma Yamada, Naoki Morita, Izuru Nagashima, Hiroshi Inoue, Yuka Inaba, Natsumi Noda, Riichiro Abe, Kazuo Umezawa, Michitaka Ozaki
ANTIOXIDANTS & REDOX SIGNALING, 21, 18, 2515, 2530, MARY ANN LIEBERT, INC, 2014年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Aims: Liver injury and regeneration involve complicated processes and are affected by various physio-pathological factors. We investigated the mechanisms of steatosis-associated liver injury and delayed regeneration in a mouse model of partial hepatectomy. Results: Initial regeneration of the steatotic liver was significantly delayed after hepatectomy. Although hepatocyte proliferation was not significantly suppressed, severe liver injury with oxidative stress (OS) occurred immediately after hepatectomy in the steatotic liver. Fas-ligand (FasL)/Fas expression was upregulated in the steatotic liver, whereas the expression of antioxidant and anti-apoptotic molecules (catalase/MnSOD/Ref-1 and Bcl-2/Bcl-xL/FLIP, respectively) and p62/SQSTM1, a steatosis-associated protein, was downregulated. Interestingly, pro-survival Akt was not activated in response to hepatectomy, although it was sufficiently expressed even before hepatectomy. Suppression of p62/SQSTM1 increased FasL/Fas expression and reduced nuclear factor erythroid 2-related factor-2 (Nrf-2)-dependent antioxidant response elements activity and antioxidant responses in steatotic and nonsteatotic hepatocytes. Exogenously added FasL induced severe cellular OS and necrosis/apoptosis in steatotic hepatocytes, with only the necrosis being inhibited by pretreatment with antioxidants, suggesting that FasL/Fas-induced OS mainly leads to necrosis. Furthermore, p62/SQSTM1 re-expression in the steatotic liver markedly reduced liver injury and improved liver regeneration. Innovation: This study is the first which demonstrates that reduced expression of p62/SQSTM1 plays a crucial role in posthepatectomy acute injury and delayed regeneration of steatotic liver, mainly via redox-dependent mechanisms. Conclusion: In the steatotic liver, reduced expression of p62/SQSTM1 induced FasL/Fas overexpression and suppressed antioxidant genes, mainly through Nrf-2 inactivation, which, along with the hypo-responsiveness of Akt, caused posthepatectomy necrotic/apoptotic liver injury and delayed regeneration, both mainly via a redox-dependent mechanism. Antioxid. Redox Signal. 21, 2515-2530.

"光"を利用した新たな細胞・臓器保存法の開発
芳賀 早苗, 小澤 岳昌, 野田 なつみ, 尾崎 倫孝
Organ Biology, 21, 3, 42, 42, (一社)日本臓器保存生物医学会, 2014年10月
日本語

高血糖および脂肪化状態が肝虚血/再灌流傷害に及ぼす影響　　　　　　　　　　　　　　　
芳賀 早苗, 小澤 岳昌, 森田 直樹, 野田 なつみ, 尾崎 倫孝
日本生化学会大会プログラム・講演要旨集, 87回, [2P, 408], (公社)日本生化学会, 2014年10月
日本語

細胞・臓器移植における基礎的研究の最前線 光を応用した移植細胞機能・細胞環境のモニタリングと制御の試み
尾崎 倫孝, 芳賀 早苗, 野田 なつみ, 森田 直樹, 山田 勇磨, 小澤 岳昌
日本外科学会雑誌, 115, 臨増2, 168, 168, (一社)日本外科学会, 2014年03月
日本語

臓器保存 光学技術を応用した新たな臓器保存法、細胞・臓器移植法の開発
尾崎 倫孝, 芳賀 早苗, 森田 直樹, 野田 なつみ, 山田 勇馬, 小澤 岳昌
Organ Biology, 20, 3, 40, 40, (一社)日本臓器保存生物医学会, 2013年10月
日本語

Contribution of Toll-Like Receptor 2 to the Innate Response against Staphylococcus aureus Infection in Mice
Yimin, Masashi Kohanawa, Songji Zhao, Michitaka Ozaki, Sanae Haga, Guangxian Nan, Yuji Kuge, Nagara Tamaki
PLoS ONE, 8, 9, e74287, 2013年09月13日, ［査読有り］
英語, 研究論文（学術雑誌）, Staphylococcus aureus is a common pathogen that causes a wide range of infectious diseases. The function of TLRs, specifically TLR2, during S. aureus infection is still debated. In this study, we investigated the extent to which TLR2 contributes to the host innate response against the bacterial infection using TLR2-deficient mice. Intravenous inoculation with S. aureus resulted in all TLR2-deficient mice dying within 4 d, along with a high bacterial burden in the livers. However, histological examination showed the same degree of macrophage and neutrophil accumulation in the livers of infected TLR2-deficient mice as that in infected wild-type (WT) mice. TLR2-deficient mouse macrophages also showed normal phagocytic activity, although they failed to express CD36 that appeared on the surface of WT mouse cells upon challenge with heat-killed S. aureus. These data indicate that TLR2, as well as CD36, does not directly affect S. aureus clearance and that CD36 expression on macrophages depends on the presence of TLR2. In vivo infection with S. aureus caused significantly elevated production of TNF-α and IL-6 in the livers and blood of TLR2-deficient mice compared with those in WT mice, while the hepatic and serum levels of IL-10 decreased in these mice. In contrast, lower expression of IL-6 and IL-10, but not of TNF-α, at both the gene and protein levels was found in TLR2-deficient mouse macrophages compared to that in WT mouse cells, in response to challenge with heat-killed S. aureus. These findings suggest that the S. aureus-induced pro-inflammatory cytokine response is not dependent on macrophages and that TLR2 deficiency results in decreased IL-10 release by macrophages, which contributes to dysregulated cytokine balance, impaired bacterial clearance, and mouse death. Therefore, TLR2 possesses a protective function during S. aureus infection by regulating pro- and anti-inflammatory cytokine responses. © 2013 Yimin et al.

光応答性ルシフェラーゼによる生きた組織下でのpH時間変化モニタリング法の開発　　　　　　　　　　　　　　　
服部 満, 芳賀 早苗, 高倉 栄男, 尾崎 倫孝, 小澤 岳昌
日本分析化学会講演要旨集, 62年会, 42, 42, (公社)日本分析化学会, 2013年08月
日本語

Sustained accurate recording of intracellular acidification in living tissues with a photo-controllable bioluminescent protein
Mitsuru Hattori, Sanae Haga, Hideo Takakura, Michitaka Ozaki, Takeaki Ozawa
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 110, 23, 9332, 9337, NATL ACAD SCIENCES, 2013年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Regulation of an intracellular acidic environment plays a pivotal role in biological processes and functions. However, spatiotemporal analysis of the acidification in complex tissues of living subjects persists as an important challenge. We developed a photo-inactivatable bioluminescent indicator, based on a combination of luciferase-fragment complementation and a photoreaction of a light, oxygen, and voltage domain from Avena sativa Phototropin1 (LOV2), to visualize temporally dynamic acidification in living tissue samples. Bioluminescence of the indicator diminished upon light irradiation and it recovered gradually in the dark state thereafter. The recovery rate was remarkably sensitive to pH changes but unsusceptible to fluctuation of luciferin or ATP concentrations. Bioluminescence imaging, taken as an index of the recovery rates, enabled long-time recording of acidification in apoptotic and autophagous processes in a cell population and an ischemic condition in living mice. This technology using the indicator is widely applicable to sense organelle-specific acidic changes in target biological tissues.

A Method for Intravital Monitoring of Human Cells Using a Far-Red Luminescent Probe in Graft-Versus-Host Disease Model Mice
Hongjiang Qiao, Riichiro Abe, Nao Saito, Yasuyuki Fujita, Inkin Hayashi-Ujiie, Gang Wang, Sanae Haga, Chun Wu, Yoshihiro Ohmiya, Michitaka Ozaki, Hiroshi Shimizu
JOURNAL OF INVESTIGATIVE DERMATOLOGY, 133, 3, 841, 843, NATURE PUBLISHING GROUP, 2013年03月, ［査読有り］
英語

Immunosuppressive effects of DTCM-G, a novel inhibitor of the mTOR downstream signaling pathway
Susumu Shibasaki, Kenichiro Yamashita, Ryoichi Goto, Kenji Wakayama, Yusuke Tsunetoshi, Masaaki Zaitsu, Rumi Igarashi, Sanae Haga, Michitaka Ozaki, Kazuo Umezawa, Satoru Todo
Transplantation, 95, 4, 542, 550, 2013年02月27日, ［査読有り］
英語, 研究論文（学術雑誌）, Background: A newly developed compound, 3-[ (dodecylthiocarbonyl)methyl]- glutarimide (DTCM-G), has been shown to inhibit nuclear translocation of c-Fos/c-Jun in a murine macrophage cell line. Herein, we studied the immunosuppressive properties and potency of DTCM-G. METHODS: Using purified mouse T cells, the in vitro effects of DTCM-G on activation, cytokine production, proliferation, and cell cycle progression were assessed, and a possible molecular target of DTCM-G was investigated. In a BALB/c (H-2) to C57BL/6 (H-2) mouse heart transplantation model, transplant recipients were administered DTCM-G, a calcineurin inhibitor (tacrolimus), and a nuclear factor-κB inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ). Treatment drugs were administered daily for 14 days after transplantation. Alloimmune responses were assessed in addition to graft survival time. RESULTS: After anti-CD3+anti-CD28 monoclonal antibody stimulation, DTCM-G significantly suppressed proliferation, interferon-γ production, and cell cycle progression of activated T cells but not CD25 expression or interleukin-2 production. These effects were accompanied by inhibition of 70-kDa S6 protein kinase phosphorylation, a downstream kinase of the mammalian target of rapamycin. The addition of tacrolimus and DHMEQ to DTCM-G resulted in a robust inhibition of T-cell proliferation. In vivo combination therapy of DTCM-G plus either tacrolimus or DHMEQ significantly suppressed alloreactive interferon-γ-producing precursors and markedly prolonged cardiac allograft survival. Furthermore, combination of all three agents markedly inhibited alloimmune responses and permitted long-term cardiac allograft survival. Conclusions: DTCM-G inhibits T cells by suppressing the downstream signal of mammalian target of rapamycin. DTCM-G in combination with tacrolimus and DHMEQ induces a strong immunosuppressive effect in vivo. © 2013 by Lippincott Williams &
Wilkins.

Anoikis induction and inhibition of peritoneal metastasis of pancreatic cancer cells by a nuclear factor-κB inhibitor, (-)-DHMEQ.
Sato M, Nakanishi K, Haga S, Fujiyoshi M, Baba M, Mino K, Yimin, Niwa H, Yokoo H, Umezawa K, Ohmiya Y, Kamiyama T, Todo S, Taketomi A, Ozaki M
Oncology research, 21, 6, 333, 343, COGNIZANT COMMUNICATION CORP, 2013年, ［査読有り］
英語, 研究論文（学術雑誌）, The transcription factor nuclear factor-kappa B (NF-kappa B) plays a crucial role in pancreatic cancer (PC) progression. NF-kappa B is also involved in resistance to anoikis, a special type of apoptosis induced when cells are detached from the extracellular matrix or other cells. Anoikis resistance is related to the metastatic abilities of tumor cells; however, little is known about anoikis induction as it relates to inhibition of PC metastasis by NF-kappa B inhibitors. Here we used a specific NF-kappa B inhibitor, (-)-dehydroxymethylepoxyquinomicin (DHMEQ), to investigate anoikis induction and peritoneal metastasis suppression following NF-kappa B inhibition. We transduced Glue, a secretory form of luciferase, into a PC cell line, AsPC-1 (AsPC-1-Gluc), for our in vivo experiments. (-)-DHMEQ induced anoikis in AsPC-1-Gluc cells as measured by cell survival assays and flow cytometry. The DNA-binding activity of p65 was enhanced immediately after cell detachment from culture dishes in ELISA assays. Some antiapoptotic proteins such as cellular inhibitor of apoptotic protein-1 were consequently upregulated on Western blots. (-)-DHMEQ prevented this increase in p65 activity and the subsequent expressions of antiapoptotic molecules. In a murine xenograft model, anoikis-resistant PC cell lines tended to metastasize to the peritoneum more than anoikis-sensitive cells, suggesting a correlation between anoikis sensitivity and peritoneal metastasis. (-)-DHMEQ successfully inhibited peritoneal metastasis of AsPC-1-Gluc cells. We monitored metastasis inhibition by ex vivo chemiluminescent detection of Glue secreted from tumor cells into murine plasma and by in vivo imaging. Our results suggest that (-)-DHMEQ inhibited peritoneal dissemination by preventing anoikis resistance of PC cells.

Successful transplantation of rat hearts subjected to extended cold preservation with a novel preservation solution
Kenji Wakayama, Moto Fukai, Kenichiro Yamashita, Taichi Kimura, Gentaro Hirokata, Susumu Shibasaki, Daisuke Fukumori, Sanae Haga, Mitsuru Sugawara, Tomomi Suzuki, Masahiko Taniguchi, Tsuyoshi Shimamura, Hiroyuki Furukawa, Michitaka Ozaki, Toshiya Kamiyama, Satoru Todo
TRANSPLANT INTERNATIONAL, 25, 6, 696, 706, WILEY-BLACKWELL, 2012年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Since prolonged cold preservation of the heart deteriorates the outcome of heart transplantation, a more protective preservation solution is required. We therefore developed a new solution, named Dsol, and examined whether Dsol, in comparison to UW, could better inhibit myocardial injury resulting from prolonged cold preservation. Syngeneic heterotopic heart transplantation in Lewis rats was performed after cold preservation with UW or Dsol for 24 or 36 h. In addition to graft survival, myocardial injury, ATP content, and Ca2+ -dependent proteases activity were assessed in the 24-h preservation group. The cytosolic Ca2+ concentration of H9c2 cardiomyocytes after 24-h cold preservation was assessed. Dsol significantly improved 7-day graft survival after 36-h preservation. After 24-h preservation, Dsol was associated with significantly faster recovery of ATP content and less activation of calpain and caspase-3 after reperfusion. Dsol diminished graft injury significantly, as revealed by the lower levels of infarction, apoptosis, serum LDH and AST release, and graft fibrosis at 7-day. Dsol significantly inhibited Ca2+ overload during cold preservation. Dsol inhibited myocardial injury and improved graft survival by suppressing Ca2+ overload during the preservation and the activation of Ca2+ -dependent proteases. Dsol is therefore considered a better alternative to UW to ameliorate the outcome of heart transplantation.

Dendritic Cells Conditioned With NK026680 Prolong Cardiac Allograft Survival in Mice
Susumu Shibasaki, Kenichiro Yamashita, Yoshiki Yanagawa, Ryoichi Goto, Kenji Wakayama, Gentaro Hirokata, Yusuke Tsunetoshi, Masaaki Zaitsu, Rumi Igarashi, Sanae Haga, Michitaka Ozaki, Satoru Todo
TRANSPLANTATION, 93, 12, 1229, 1237, LIPPINCOTT WILLIAMS & WILKINS, 2012年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Background. Pharmacologically modulated dendritic cells (DCs) can potentially regulate alloimmune responses. We examined the characteristics of immunoregulatory DCs induced by a novel triazolopyrimidine derivative, NK026680, which has been previously shown to inhibit DC maturation.
Methods. DCs were generated from bone marrow progenitor cells from C57BL/6 (B6, H-2(b) haplotype) mice with granulocyte-macrophage colony-stimulating factor and interleukin (IL)-4. DCs were cultured with allogeneic BALB/c (H-2(d)) splenocyte lysates with or without NK026680. DC functions were examined in vitro after stimulation of tumor necrosis factor alpha and in vivo by the intravenous injection of C3He/J (C3H, H-2(k)) DCs cultured with B6 cell lysates and NK026680 into C3H mice. Seven days later, DC-treated mice received B6 heart allografts, and graft survival and alloimmune responses were assessed.
Results. In NK026680-treated DCs (NK-DCs), significant inhibition of the up-regulation of surface activation markers (CD40, CD80, CD86, and major histocompatibility complex class II) and IL-12 p40 production was observed after stimulation of tumor necrosis factor alpha compared with that of control DCs. Furthermore, NK-DCs suppressed alloreactive T-cell proliferation. The modulation of NK-DCs was likely associated with the inhibition of phosphorylation of p38 mitogen-activated protein kinase and the up-regulation of indolamine 2,3-dioxygenase expression. Compared with both noninjected and control DC-injected mice, mice that received a single in vivo infusion of NK-DCs showed significant increases in splenocyte IL-10 production and the splenic CD4(+) IL-10(high) T-cell population 7 days after injection, a significantly increased splenic CD4(+)CD25(+)FoxP3(+) T-cell population 14 days after injection, and markedly prolonged cardiac allograft survival.
Conclusions. Ex vivo NK026680 conditioning allows DCs to acquire immunoregulatory properties that suppress alloimmune responses and prolong cardiac allograft survival.

Bio-imaging of surgical stress: Dynamic analysis of liver oxidative stress and damage
Morita Naoki, Haga Sanae, Ozawa Takeaki, Remington S. James, Ozaki Michitaka
LUMINESCENCE, 27, 2, 143, 145, 2012年03月, ［査読有り］

NK026680 inhibits T-cell function in an IL-2-dependent manner and prolongs cardiac allograft survival in rats
Susumu Shibasaki, Kenichiro Yamashita, Ryoichi Goto, Tetsu Oura, Kenji Wakayama, Gentaro Hirokata, Tomohiro Shibata, Rumi Igarashi, Sanae Haga, Michitaka Ozaki, Satoru Todo
TRANSPLANT IMMUNOLOGY, 26, 1, 42, 49, ELSEVIER SCIENCE BV, 2012年01月, ［査読有り］
英語, 研究論文（学術雑誌）, NK026680 is a triazolopyrimidine derivative that has been shown to inhibit dendritic cell maturation and activation. Here, we examined the immunosuppressive properties of NK026680 on T-cell function and assessed its immunosuppressive efficacy in an ACI (RT1(av1) haplotype) to Lewis (RT1(I)) rat heart transplantation model. The effects of NK026680 on T-cell proliferation, activation, and cytokine production were investigated in vitro. Heart transplant recipient rats were administered NK026680 daily for 14 days post-transplantation. In addition to graft survival time, alloimmune responses and graft histology at 4-10 days post-transplantation were assessed. NK026680 was found to inhibit proliferation, CD25 upregulation, IL-2 production, and cell cycle progression in alpha CD3/alpha CD28-stimulated murine T cells. These effects were likely due to suppression of the p38 mitogen-activated protein kinase pathway and the subsequent inhibition of p65, c-Fos, and to a lesser extent, c-Jun. Daily NK026680 treatment suppressed alloimmune responses, prevented cellular infiltration into allografts, and prolonged graft survival. The anti-rejection effects of NK026680 were enhanced by tacrolimus. In conclusion, NK026680 inhibits the activation of T cells and prolongs cardiac allograft survival in rats. These features make it a potential candidate immunosuppressant for the treatment of organ transplant patients in the future. (C) 2011 Elsevier B.V. All rights reserved.

In Vivo Monitoring of Liver Damage Using Caspase-3 Probe
Michitaka Ozaki, Sanae Haga, Takeaki Ozawa
THERANOSTICS, 2, 2, 207, 214, IVYSPRING INT PUBL, 2012年, ［査読有り］
英語, 研究論文（学術雑誌）, Real-time monitoring of cellular and organ conditions improves our understanding of various physiopathological phenomena. Such monitoring is expected to provide important alternatives for clinical diagnosis and therapy. We have sought to show physiopathological changes of organs as well as cells. Here, we present an example of in vivo imaging of liver states using the luciferase-based caspase-3 optical probe. We examined dynamic changes of apoptosis (caspase-3 activity) of a mouse liver as well as those of liver cells, proving that the emitted signals reflected the biochemically evaluated apoptotic cell death. In live liver cell (AML 12) experiments, the optical probe for caspase-3 activity emitted signals in response to Fas-ligand, staurosporine and hypoxia/reoxygenation, demonstrating that the probe can measure cellular apoptosis quantitatively. We therefore applied this probe for mouse liver ischemia/reperfusion (I/R) and drug-toxicity to liver. By expressing the probe in a mouse liver adenovirally, we imaged liver caspase-3 activity (i.e. apoptotic damage) non-invasively and chronologically in the hepatic I/R model of mice. The duration of liver ischemia affected the post-ischemic caspase-dependent damage. Ischemia (up to 60 min) enhanced liver damage after reperfusion, but prolonged ischemia (90 min of ischemia) induced not apoptotic cell death but necrotic cell death. Direct observations of the changes of organ conditions elucidated the dynamism of organ function and damage. These technologies clearly possess clinical relevance. They are expected to provide a new diagnostic tool for various clinical settings in the future.

'Collagen vitrigel sheet' as a novel drug delivery bio-material
S. Haga, S. Haga, T. Takezawa, T. Ozawa, S. J. Remington, N. Morita, M. Ozaki
IFMBE Proceedings, 37, 1354, 1357, 2011年11月09日
Background/Aims. Ischemia/reperfusion (I/R)- induced injury is an acute injury mainly caused by surgical procedures. And HGF (hepatocyte growth factor) is known to protect against I/R-injury effectively. The sufficient dose of HGF must be topically delivered to the target organ to minimize the organ damage on surgery (reperfusion) and the dose of drug. For this purpose, we developed a novel technology for converting a fragile disk of the conventional type-I collagen gel into a strong and transparent 'vitrigel membrane', based on a concept for the vitrification of heat-denatured proteins. In the present study, we examined the efficacy of the newly designed 'HGF-containing collagen vitrigel sheet' as an effective drug delivery system in mouse liver I/R model. Materials and methods. Male C57Black6 mice were subjected to 60 min-liver ischemia (middle and left liver lobes) and reperfusion. Immediately after liver ischemia, the ischemic liver lobes were covered by collagen vitrigel sheet containing hHGF (3..g/ml). Reperfused liver injury was evaluated by biochemical/ histological methods and bio-imaging of hepatic oxidative stress (redox-sensitive GFP probe) and apoptosis (bioluminescent caspase-3 activity probe). Results. Collagen vitrigel sheet (containing hHGF) released hHGF until 96 hr in vitro and successfully delivered hHGF to the liver tissue suffering I/R. This effectively protected liver from I/R-induced injury. Blood biochemistry showed reduced levels of serum GOT/GPT/LDH, and liver histology revealed reduced necrotic area in the hHGF-treated liver. Oxidative stress and caspase-3 activity of the liver were examined by bioimaging, which revealed that hHGF delivered by vitrigel sheet significantly reduced oxidative stress and apoptotic cell death in the post-ischemic liver. Conclusions. hHGF-containing collagen vitrigel sheet protected liver from I/R-induced injury. Collagen vitrigel sheet may have good therapeutic potential in topical drug delivery in various clinical settings. © 2011 Springer-Verlag Berlin Heidelberg.

AGE-ASSOCIATED P66SHC INDUCED REDOX-DEPENDENT LIVER DAMAGE AND IMPAIRED REGENERATION AFTER HEPATECTOMY IN AGED MICE
Ozaki Michitaka, Haga Sanae, Irani Kaikobad, Morita Naoki, Kaneshima Yoshimi, Ozawa Takeaki
HEPATOLOGY, 54, 686A, 687A, 2011年10月, ［査読有り］

Inhibition of nuclear factor-kappaB suppresses peritoneal dissemination of gastric cancer by blocking cancer cell adhesion
Kazuhiro Mino, Michitaka Ozaki, Kazuaki Nakanishi, Sanae Haga, Masanori Sato, Masaya Kina, Masato Takahashi, Norihiko Takahashi, Akihiko Kataoka, Kazuyoshi Yanagihara, Takahiro Ochiya, Toshiya Kamiyama, Kazuo Umezawa, Satoru Todo
CANCER SCIENCE, 102, 5, 1052, 1058, WILEY-BLACKWELL, 2011年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Currently, patients with peritoneal dissemination of gastric cancer must accept a poor prognosis because there is no standard effective therapy. To inhibit peritoneal dissemination it is important to inhibit interactions between extracellular matrices (ECM) and cell surface integrins, which are important for cancer cell adhesion. Although nuclear factor-kappa B (NF-kappa B) is involved in various processes in cancer progression, its involvement in the expression of integrins has not been elucidated. We used a novel NF-kappa B inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ), to study whether NF-kappa B blocks cancer cell adhesion via integrins in a gastric cancer dissemination model in mice and found that DHMEQ is a potent suppressor of cancer cell dissemination. Dehydroxymethylepoxyquinomicin suppressed the NF-kappa B activity of human gastric cancer cells NUGC-4 and 44As3Luc and blocked the adhesion of cancer cells to ECM when compared with the control. Dehydroxymethylepoxyquinomicin also inhibited expression of integrin (alpha 2, alpha 3, beta 1) in in vitro studies. In the in vivo model, we injected 44As3Luc cells pretreated with DHMEQ into the peritoneal cavity of mice and performed peritoneal lavage after the injection of cancer cells. Viable cancer cells in the peritoneal cavities were evaluated sequentially by in vivo imaging. In mice injected with DHMEQ-pretreated cells and lavaged, live cancer cells in the peritoneum were significantly reduced compared with the control, and these mice survived longer. These results indicate that DHMEQ could inhibit cancer cell adhesion to the peritoneum possibly by suppressing integrin expression. Nuclear factor-kappa B inhibition may be a new therapeutic option for suppressing postoperative cancer dissemination. (Cancer Sci 2011; 102: 1052-1058).

Bio-imaging of surgical stresses - Dynamic analyses of liver oxidative stress and damage - Dynamic a
M. Ozaki, S. Haga, T. Ozawa, N. Morita, Y. Kaneshima, J. Remington
IFMBE Proceedings, 37, 1094, 1097, 2011年
英語, 研究論文（国際会議プロシーディングス）, Real-time monitoring of cellular and organ conditions lead to better understandings of various physiopathological phenomena, and will provide options in clinical diagnosis and therapy. We developed redox sensitive GFP (roGFP) and luciferase-based caspase-3 optical probes for in vivo imaging. We tried to visualize the dynamic changes of oxidative stress and the following damage in live cells and in mouse liver ischemia/reperfusion (I/R) model. In live cell experiments, roGFP probe visualized realtime oxidation/reduction (redox) changes induced by hypoxia/ reoxygenation (H/R). On reoxygenation, roGFP successfully illustrated post-hypoxic cellular oxidation and the following recovery to the reductive basal condition. The optic probe for Caspase-3 activity showed signals in response to Fas-ligand challenge to AML12 liver cells, which paralleled to cellular apoptosis. In the hepatic I/R model of mice, we successfully imaged liver oxidative stress (by roGFP probe) and apoptosis (by caspase-3 activity probe) non-invasively and chronologically in a single mouse. Duration of liver ischemia affected the intensity of post-ischemic oxidative stress and caspase-dependent apoptotic cell death (damage). Prolonged ischemia (until 60 min) enhanced liver oxidative stress and damage after reperfusion. Interestingly, 90 min of ischemia did not induce oxidative stress at all, but necrotic cell death. Direct observations of the changes of organ conditions helped us understand the dynamism of organ functions/ conditions. These technologies clearly possess a clinical relevance and will provide a new diagnostic tool in various clinical settings in the future. © 2011 Springer-Verlag Berlin Heidelberg.

p66(Shc) has a pivotal function in impaired liver regeneration in aged mice by a redox-dependent mechanism
Sanae Haga, Naoki Morita, Kaikobad Irani, Masato Fujiyoshi, Tetsuya Ogino, Takeaki Ozawa, Michitaka Ozaki
LABORATORY INVESTIGATION, 90, 12, 1718, 1726, NATURE PUBLISHING GROUP, 2010年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Liver regeneration involves complicated processes and is affected by various patho-physiological conditions. This study was designed to examine the molecular mechanisms underlying the aging-associated impairment of liver regeneration. Male C57BL/6J mice were used as young and aged mice (<10 weeks and >20 months old, respectively). These mice were subjected to 70% partial hepatectomy (PH). Liver regeneration and liver injury/stresses were evaluated chronologically after PH. Post-hepatectomy liver regeneration was markedly impaired in aged mice. Though the extent of hepatocyte proliferation in the regenerating liver was similar in aged and young mice, cell growth was absent in aged mice. Oxidative stress (OS) was observed immediately after hepatectomy, followed by marked apoptosis in aged mice. Signaling molecules regarding cell proliferation (mitogen-activated protein kinase, STAT3, p46/52(Shc)) and anti-oxidation (catalase, superoxide dismutase, Ref-1, glutathione peroxidase) were expressed/activated after hepatectomy in livers of both aged and young mice. Akt was not activated in aged-mouse liver, but its expression was similar to that in young mice. p66(Shc), known as an age-/oxidant-associated protein, was strongly phosphorylated. By knocking down p66(Shc), the impairment of liver regeneration was normalized. OS immediately after hepatectomy induced subsequent liver injury (apoptosis), and deletion of p66(Shc) suppressed both OS and hepatocyte apoptosis in the regenerating liver of aged mice. Though we need additional data in other animal models to fully understand the mechanism, p66(Shc) may have a pivotal function in the impairment of liver regeneration in aged mice by triggering OS and subsequent apoptosis. This data may provide a clue to understanding the mechanism underlying the association between aging and the impairment of liver regeneration. Laboratory Investigation (2010) 90, 1718-1726; doi:10.1038/labinvest.2010.119; published online 21 June 2010

[Development of novel cell culture systems utilizing the advantages of collagen vitrigel membrane].
Toshiaki Takezawa, Maya Fukuda, Winnette McIntosh-Ambrose, Ji-Ae Ko, Jennifer Elisseeff, Sanae Haga, Michitaka Ozaki, Kiyoko Kato, Pi-Chao Wang, Tadashi Uchino, Teruo Nishida
Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan, 130, 4, 565, 74, 2010年04月, ［査読有り］, ［国内誌］
日本語, 研究論文（学術雑誌）, The white of an egg, rendered opaque by boiling, can be converted into a thin, transparent and rigid material like glass by evaporating the moisture. This phenomenon is known as the vitrification of heat-denatured proteins. We applied vitrification technology to a collagen gel and converted it into a rigid glass-like material. We attempted to rehydrate the glass-like material and succeeded in preparing a novel stable state of collagen gel that was a thin and transparent membrane with excellent gel strength and protein permeability. We called it "collagen vitrigel" because it was produced from the vitrification process of a traditional hydrogel. Further, a framework-embedded collagen vitrigel membrane that can be easily turned inside out with tweezers was prepared by inserting a nylon membrane ring in the collagen sol prior to the gelation, thereby allowing the membrane to function as a removable cell culture substratum. Different types of anchorage-dependent cells could be cultured on both surfaces of the substratum by the manipulation of two-dimensional cultures, and consequently a three-dimensional crosstalk model with paracrine effects from each cell type was reconstructed. Also, the collagen vitrigel membrane containing a bioactive molecule provided a drug delivery system (DDS) with sustainable release. In this review, we summarize the recent progress of applied studies using the collagen vitrigel membrane as follows: a corneal model for eye irritant and permeability tests, a skin model for sensitization test, a renal glomerular model for evaluating blood filtration, an endometrial model for developing a new treatment and a DDS of hepatocyte growth factor for improving liver disorder.

Immunosuppressive Effects of Regulatory Dendritic Cells Induced by NK026680.
Susumu Shibasaki, Kenichiro Yamashita, Ryoichi Goto, Kenji Wakayama, Gentaro Hirokata, Yusuke Tsunetoshi, Masaaki Zaitsu, Rumi Igarashi, Sanae Haga, Yoshiki Yanagawa, Michitaka Ozaki, Satoru Todo
AMERICAN JOURNAL OF TRANSPLANTATION, 10, 560, 561, WILEY-BLACKWELL PUBLISHING, INC, 2010年04月, ［査読有り］
英語

HEPATIC ISCHEMIA INDUCED IMMEDIATE OXIDATIVE STRESS AFTER REPERFUSION AND DETERMINED THE SEVERITY OF THE REPERFUSION-INDUCED DAMAGE. - IN VIVO DYNAMIC ANALYSIS OF LIVER OXIDATIVE STRESS AND DAMAGE
Ozaki Michitaka, Haga Sanae, Remington S. James, Morita Naoki, Ozawa Takeaki
HEPATOLOGY, 50, 4, 640A, 2009年10月, ［査読有り］

Hepatic Ischemia Induced Immediate Oxidative Stress after Reperfusion and Determined the Severity of the Reperfusion-Induced Damage
Sanae Haga, S. James Remington, Naoki Morita, Keita Terui, Michitaka Ozaki
ANTIOXIDANTS & REDOX SIGNALING, 11, 10, 2563, 2572, MARY ANN LIEBERT, INC, 2009年10月, ［査読有り］
英語, 研究論文（学術雑誌）, Noninvasive evaluation of organ redox states provides invaluable information in many clinical settings. We evaluated a newly developed reduction/oxidation-sensitive green fluorescent protein (roGFP) probe that reports cellular redox potentials and their dynamic changes in live cells. On hypoxia/reoxygenation (H/R) of AML12 liver cells, roGFP indicated mild reduction during hypoxia, but immediate transient oxidation after reoxygenation. The roGFP probe confirmed the antioxidative effects of N-acetylcysteine, catalase, redox factor-1, and Mn-SOD/CuZn-SOD against H/R-induced cellular oxidative stress (OS). In a mouse liver ischemia/reperfusion (I/R) model, roGFP transduced by using an adenoviral vector revealed immediate reduction of the liver under ischemia, and two distinct peaks of OS: (a) early, observed within 60 min after reperfusion, similar to the in vitro study; and (b) later, at 24 h. The early peak levels paralleled the ischemic time up to 75 min and the postischemic liver injury (sGOT/GPT/LDH) in the later phase (6 and 24 h after I/R). The roGFP probe successfully indicated postischemic OS of the liver in living mice, accurately predicting postischemic liver injury. This probe may represent an effective OS marker indicating organ redox states and also predicting the damage/function. Antioxid. Redox Signal. 11, 2563-2572.

マウスモデルにおける新規NF-kB阻害剤DHMEQの膵癌腹膜播種抑制効果(A novel NF-κB inhibitor DHMEQ suppressed pancreas cancer progression in a mouse model of peritoneal dissemination)　　　　　　　　　　　　　　　
佐藤 正法, 尾崎 倫孝, 中西 一彰, 渡邉 俊之, 三野 和宏, 芳賀 早苗, 横尾 英樹, 梅澤 一夫, 近江谷 克祐, 神山 俊哉, 藤堂 省
日本癌学会総会記事, 68回, 481, 481, 日本癌学会, 2009年08月
英語

The Survival Pathways Phosphatidylinositol-3 Kinase (PI3-K)/Phosphoinositide-Dependent Protein Kinase 1 (PDK1)/Akt Modulate Liver Regeneration Through Hepatocyte Size Rather Than Proliferation
Sanae Haga, Michitaka Ozaki, Hiroshi Inone, Yasuo Okamoto, Wataru Ogawa, Kiyoshi Takeda, Shizuo Akira, Satoru Todo
HEPATOLOGY, 49, 1, 204, 214, JOHN WILEY & SONS INC, 2009年01月, ［査読有り］
英語, 研究論文（学術雑誌）, Liver regeneration comprises a series of complicated processes. The current study was designed to investigate the roles of phosphoinositide-dependent protein kinase 1 (PDK1)-associated pathways in liver regeneration after partial hepatectomy (PH) using liver-specific Pdk1-knockout (L-Pdk1KO) and Pdk1/STAT3 double KO (L-DKO) mice. There was no liver regeneration, and 70% PH was lethal in L-Pdk1KO mice. Liver regeneration was severely impaired equally in L-Pdk1KO and L-DKO mice, even after nonlethal 30% PH. There was no cell growth (measured as increase of cell size) after hepatectomy in L-Pdk1KO mice, although the post-PH mitotic response was the same as in controls. As expected, hepatectomy did not induce hepatic Akt-phosphorylation (Thr308) in L-Pdk1KO mice, and post-PH phosphorylation of Akt, mammalian target of rapamycin (mTOR), p70 ribosomal S6 kinase (P70(S6K)), and S6 were also reduced. To examine the specific role of PDK1-associated signals, a "pif-pocket" mutant of PDK1, which allows PDK1 only to phosphorylate Akt, was used. Liver regeneration was recovered in L-Pdk1KO mice with a "pif-pocket" mutant of PDK1. This re-activated Akt in L-Pdk1KO mice liver and induced post-PH cell growth, without affecting cell proliferation. Further deletion of STAT3 (L-DKO mice) did not further deteriorate liver regeneration, although this certainly reduced post-PH mitotic response. These findings indicate that PDK1/Akt contribute to liver regeneration by regulating cell size. Regarding phosphatidylinositol-3 kinase (PI3-K), immediate upstream signal of PDK1, activation of PI3-K induced cell proliferation via STAT3 activation in the liver of L-Pdk1KO mice but did not improve impaired liver regeneration. This confirmed the pivotal role of PDK1 in liver regeneration and cell growth. Conclusion: PDK1/Akt-mediated responsive cell growth is essential for normal liver regeneration after PH, especially when cell proliferation is impaired. (HEPATOLOGY 2009;49:204-214.)

Mutual modulation between interleukin-10 and interleukin-6 induced by Rhodococcus aurantiacus infection in mice
Yimin, Masashi Kohanawa, Michitaka Ozaki, Sanae Haga, Keiko Fujikawa, Songji Zhao, Yuji Kuge, Nagara Tamaki
MICROBES AND INFECTION, 10, 14-15, 1450, 1458, ELSEVIER SCIENCE BV, 2008年11月, ［査読有り］
英語, 研究論文（学術雑誌）, The interaction between interleukin-10 (IL-10) and interleukin-6 (IL-6) was investigated in the inflammatory response to Rhodococcus aurantiacus (R. aurantiacus) infection, in which both cytokines act as anti-inflammatory cytokines. Compared with wild-type (WT) counterparts, IL-6 gene-deficient (IL-6(-)/(-)) mice mounted a more robust production of IL-10 and tumor necrosis factor-alpha (TNF-alpha) during the initial phase of infection. Administration of anti-IL-10 antibody resulted in all the mice dying within 3 days post-infection as well as a further elevated TNF-alpha release. In vitro challenge of the macrophages from IL-6(-)/(-) and WT mice with heat-killed R. aurantiacus also showed similar results. Addition of exogenous IL-6 depressed IL-10 and TNF-alpha production by either IL-6(-)/(-) mice or IL-6(-)/(-) mouse macrophages. Likewise, WT mouse macrophages pretreated with anti-IL-10 or anti-IL-6 antibody exhibited increased production of TNF-alpha and IL-6 or IL-10 respectively. Moreover, neutralization of both IL-10 and IL-6 induced a further increase in TNF-alpha production by WT mouse cells. Overall, we conclude that IL-10 is a key element in protecting mice against mortality, and that IL-10 and IL-6 production are negatively regulated by each other although they are additive in suppressing TNF-alpha release in R. aurantiacus-infected mouse model. (C) 2008 Elsevier Masson SAS. All rights reserved.

マウス胃癌腹膜播種モデルにおける、新規NF-κB阻害剤DHMEQの腫瘍抑制効果の検討(A novel NF-κB inhibitor DHMEQ suppresses the expansion of gastric cancer in the mouse peritoneal dissemination model)　　　　　　　　　　　　　　　
三野 和宏, 中西 一彰, 喜納 政哉, 佐藤 正法, 芳賀 早苗, 横尾 英樹, 神山 俊哉, 梅澤 一夫, 尾崎 倫孝, 藤堂 省
日本癌学会総会記事, 67回, 183, 183, 日本癌学会, 2008年09月
英語

Identification of de novo STAT3 target gene in liver regeneration
Hui-Qi Zhang, Sanae Haga, Moto Fukai, Yuko Oikawa, Hiroshi Inoue, Wataru Ogawa, Arihiko Kano, Atsushi Maruyama, Xin-Yuan Fu, Satoru Todo, Shin Enosawa, Michitaka Ozaki
Hepatology Research, 38, 4, 374, 384, 2008年04月, ［査読有り］
英語, 研究論文（学術雑誌）, The process of liver regeneration is regulated by complex mechanisms. Although signal transducer and activator of transcription-3 (STAT3), a transcription factor which targets mainly mitotic genes, definitely plays an important role in liver regeneration, the exact roles of STAT3 are not completely understood. Aim: In this report, we tried to search for a new target of STAT3 involved in liver regeneration in mice. Methods: We generated liver-specific STAT3 knockout (L-S3KO) mice and a STAT3 knockout cell line of mouse origin. Using chromatin immunoprecipitation, we screened 12 genes to which STAT3 binds after partial hepatectomy (PH). Of these genes, we analyzed the S3-IE3 clone that is located on chromosome-3 and possesses STAT3 binding sites in it. Results: We showed that STAT3 binds to a specific site on S3-IE3, and that interleukin-6 (IL-6) stimulates its transcriptional activity. The mRNA and protein levels of the net gene, which is located downstream of S3-IE3, were negatively regulated in the control cells, but not in the STAT3 knockout cells after IL-6 stimulation. Similarly in in vivo mouse PH, the mRNA and protein levels of net were also negatively regulated after PH, but not in L-S3KO mice. Conclusion: The net gene is located downstream of a newly-recognized STAT3 binding site (S3-IE3) and negatively regulated after IL-6 stimulation and PH, although its role is still unclear. © 2007 The Japan Society of Hepatology.

Preventing hypoxia/reoxygenation damage to hepatocytes by p66(shc) ablation: Up-regulation of anti-oxidant and anti-apoptotic proteins
Sanae Haga, Keita Terui, Moto Fukai, Yuko Oikawa, Kaikobad Irani, Hiroyuki Furukawa, Satoru Todo, Michitaka Ozaki
JOURNAL OF HEPATOLOGY, 48, 3, 422, 432, ELSEVIER SCIENCE BV, 2008年03月, ［査読有り］
英語, 研究論文（学術雑誌）, Background/Aims:Ischemia/reperfusion damage to the liver remains a serious concern in many clinical situations. Major mechanisms for this certainly include oxidative stress.
Methods:The effects of ablating the p66 isoform of ShcA (p66(shc)) on hypoxia/reoxygenation (H/R)-induced oxidative stress and cell injury in hepatocytes were investigated.
Results: Immediately after reoxygenation, AML12 cells were clearly under oxidative stress; many cells underwent apoptosis. However, knockdown of p66(shc) by specific RNAi markedly decreased cellular oxidative stress and H/R-induced apoptosis, as well as conferring resistance to H2O2 insult. These data suggest that prevention of apoptosis conferred by ablation of p66(shc) results from changed ROS-scavenging, but not inhibition of ROS generation. These data were also confirmed in fibroblasts from p66(shc) knockout mice. Anti-oxidant molecules, such as MnSOD and Ref-1 and the anti-apoptotic molecule Bcl-xL were up-regulated, and pro-apoptotic FLICE was down-regulated, by ablation of p66. Interestingly, catalase expression was not affected in p66(shc)-knockdown-AML12 cells although it is a major target in other cell types.
Conclusions: Our findings suggest that in hepatocytes, ablation of p66 A, is cytoprotective against H/R-induced oxidative stress, with MnSOD and Ref-1 playing critical roles, and with up-regulation of Bcl-xL and down-regulation of FLICE contributing jointly to preventing cells from undergoing oxidant-induced apoptosis. (C) 2007 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Mechanism of impaired regeneration of fatty liver in mouse partial hepatectomy model
Hiroshi Murata, Takahito Yagi, Hiromi Iwagaki, Tetsuya Ogino, Hiroshi Sadamori, Hiroyoshi Matsukawa, Yuzoh Umeda, Sanae Haga, Noriaki Takaka, Michitaka Ozaki
JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, 22, 12, 2173, 2180, BLACKWELL PUBLISHING, 2007年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Background and Aim: The mechanism of injury in steatotic liver under pathological conditions been extensively examined. However, the mechanism of an impaired regeneration is still not well understood. The aim of this study was to analyze the mechanism of impaired regeneration of steatotic liver after partial hepatectomy (PH).
Methods: db/db fatty mice and lean littermates were used for the experiments. Following 70% PH, the survival rate and recovery of liver mass were examined. Liver tissue was histologically examined and analyzed by western blotting and RT-PCR.
Results: Of 35 db/db mice, 25 died within 48 h of PH, while all of the control mice survived. Liver regeneration of surviving db/db mice was largely impaired. In db/db mice, mitosis of hepatocytes after PH was disturbed, even though proliferating cell nuclear antigen (PCNA) expression (G1 to S phase marker) in hepatocytes was equally observed in both mice groups. Interestingly, phosphorylation of Cdc2 in db/db mice was suppressed by reduced expression of Wee1 and Myt1, which phosphorylate Cdc2 in S to G2 phase.
Conclusions: In steatotic liver, cell-cycle-related proliferative disorders occurred at mid-S phase after PCNA expression. Reduced expression of Wee1 and Myt1 kinases may therefore maintain Cdc2 in an unphosphorylated state and block cell cycle progression in mid-S phase. These kinases may be critical factors involved in the impaired liver regeneration in fatty liver.

Control of allograft rejection by applying a novel nuclear factor-kappa B inhibitor, dehydroxymethylepoxyquinomicin
Shinya Ueki, Kenichiro Yamashita, Takeshi Aoyagi, Sanae Haga, Tomomi Suzuki, Tomoo Itoh, Masahiko Taniguchi, Tsuyoshi Shimamura, Hiroyuki Furukawa, Michitaka Ozaki, Kazuo Umezawa, Satoru Todo
TRANSPLANTATION, 82, 12, 1720, 1727, LIPPINCOTT WILLIAMS & WILKINS, 2006年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Background. Nuclear factor (NF)-kappa B plays a crucial role in lymphocyte activation, proliferation, and survival. We examined the immunosuppressive effect of a newly developed NF-kappa B inhibitor, dehydroxymethylepoxyquinomicin (DHMEQ) in allotransplantation.
Methods. Purified C57BL/6 (H-2(b)) T cells were used for in vitro studies examining activation, proliferation, cytokine production and nuclear NF-kappa B and nuclear factor of activated T cells (NFAT) protein levels. A fully major histocompatibility complex incompatible BALB/c (H-2(d))-to-C57BL/6 mice cardiac transplantation model was utilized for in vivo studies. DHMEQ was given intraperitoneally to transplant recipients at a various dose starting from day 0. In some, DHMEQ was administered concomitantly with tacrolimus.
Results. DHMEQ significantly suppressed alpha CD3 + alpha CD28 monoclonal antibody-triggered T-cell proliferation, CD25/CD69 expressions, and both interleukin-2 and interferon (IFN)-gamma production in a dose-dependent fashion. DHMEQ blocked nuclear translocation of NF-kappa B but not NFAT in activated T cells. Combined treatment with DHMEQ and tacrolimus significantly suppressed T cell activation as compared to that of mono-therapy with either agent alone. Single DHMEQ treatment moderately prolonged cardiac allograft survival. Further, combination of DHMEQ plus tacrolimus markedly prolonged graft mean survival time (MST) to 59.5 days when compared to either DHMEQ (MST: 10 days) or tacrolimus (MST: 13 days) treatment alone. Such effect was associated with inhibition of mixed lymphocyte reaction against donor antigen, IFN-gamma producing splenocytes and graft cellular infiltration as examined at 5 and 12 days posttransplantation.
Conclusion. DHMEQ inhibits nuclear translocation of NF-kappa B but not NFAT inactivated T cells, and prolongs allograft survival. Blocking both NF-kappa B and NFAT by DHMEQ and tacrolimus induces potent immunosuppression, which may become a new modality in controlling allograft rejection.

Protective effects of nafamostat mesilate on liver injury induced by lipopolysaccharide in rats: Possible involvement of CD14 and TLR-4 downregulation on Kupffer cells
Hideaki Miyaso, Yoshinori Morimoto, Michitaka Ozaki, Sanae Haga, Susumu Shinoura, Yasuhiro Choda, Hiroshi Murata, Goutaro Katsuno, Kamul Huda, Hideo Takahashi, Noriaki Tanaka, Hiromi Iwagaki
DIGESTIVE DISEASES AND SCIENCES, 51, 11, 2007, 2012, SPRINGER, 2006年11月, ［査読有り］
英語, 研究論文（学術雑誌）, Nafamostat mesilate (NM) is a synthetic protease inhibitor with various biological effects. To determine its effect on liver injury related to sepsis, we investigated the effects of NM on lipopolysaccharide (LPS)-induced liver injury. Wistar rats were allocated into two groups; the NM group underwent intraperitoneal NM administration 30 min before LPS administration, and the control group underwent PBS administration. Serum AST and ALT levels were significantly decreased in NM-treated rats. Reduced levels of TNF-alpha, IL-1 beta, and IFN-gamma were observed after LPS administration in NM-treated rats. No significant differences were observed in IL-6 levels between the NM and the control group. In contrast, HGF levels were significantly increased only in control rats. NM treatment decreased protein and mRNA levels of TLR-4 and CD14. Our data suggest that NM treatment has protective effects against LPS-induced hepatotoxicity through downregulation of TLR4 and CD14 in liver, which decreased TNF-alpha, IL-1 beta, and IFN-gamma production in liver.

Ex vivo adenoviral gene transfer of constitutively activated STAT3 reduces post-transplant liver injury and promotes regeneration in a 20% rat partial liver transplant model
KASM Huda, L Guo, S Haga, H Murata, T Ogino, M Fukai, T Yagi, H Iwagaki, N Tanaka, M Ozaki
TRANSPLANT INTERNATIONAL, 19, 5, 415, 423, BLACKWELL PUBLISHING, 2006年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Signal transducer and activator of transcription-3 (STAT3) is one of the most important transcription factors for liver regeneration. This study was designed to examine the effects of constitutively activated STAT3 (STAT3-C) on post-transplant liver injury and regeneration in a rat 20% partial liver transplant (PLTx) model by ex vivo adenoviral gene transfer. Adenovirus encoding the STAT3-C gene was introduced intraportally into liver grafts and clamped for 30 min during cold preservation. After orthotopic PLTx, liver graft/body weights and serum biochemistry were monitored, and both a histological study and DNA binding assay were performed. STAT3-C protein expression and its binding to DNA in the liver graft were confirmed by Western blotting and electrophoretic mobility shift assay (EMSA), respectively. This treatment modality promoted post-Tx liver regeneration effectively and rapidly. The serum levels of alanine aminotransferase/aspartate aminotransferase (AST/ALT) and bilirubin decreased in rats with STAT3-C. However, albumin (a marker of liver function) did not. Ex vivo gene transfer of STAT3-C to liver grafts reduced post-Tx injury and promoted liver regeneration. Thus, the activation of STAT3 in the liver graft may be a potentially effective clinical strategy for improving the outcome of small-for-size liver transplantation.

Obstructive jaundice increases sensitivity to lipopolysaccharide via TLR4 upregulation: Possible involvement in gut-derived hepatocyte growth factor-protection of hepatocytes
H Miyaso, Y Morimoto, M Ozaki, S Haga, S Shinoura, Y Choda, H Iwagaki, N Tanaka
JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, 20, 12, 1859, 1866, BLACKWELL PUBLISHING, 2005年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Background: Patients with obstructive jaundice are prone to sepsis after biliary tract surgery. The present study was designed to determine the effect of biliary obstruction on cytokine responses to lipopolysaccharide (LPS).
Methods: Wister rats were allocated into two groups; the BDL group underwent bile duct ligation, followed 2 weeks later by administration of LPS into the duodenum. The control group underwent sham operation, and similarly received enteral LPS. Specimens were collected serially, and applied for the assays.
Results: Serum aspartate aminotransferase and alanine aminotransferase levels were significantly increased in BDL rats. High tumor necrosis factor alpha (TNF-alpha) and interleukin (IL)-6 levels in peripheral blood were observed 2 h after LPS administration in BDL rats. In contrast, no increases in both cytokines were noted in peripheral and portal blood in control rats. Baseline HGF levels in portal and peripheral blood in BDL rats were significantly higher than in control rats. LPS significantly increased hepatocyte growth factor (HGF) levels in portal blood and decreased in peripheral blood in BDL rats, but not in control rats. Immunohistochemical analysis revealed that BDL increased expressions of Toll-like receptor (TLR)4, CD14 and CD68 both in the small intestine and liver. Both TLR4 and CD14 mRNAs were upregulated in the small intestine and liver after LPS administration in BDL rats.
Conclusion: Obstructive jaundice and LPS stimulation induced TLR4 upregulation both in the liver and small intestine, which led to increased TNF-alpha and IL-6 production in liver and HGF production in the small intestine. The upregulation of TLR4 may lead to pathological and host defense reactions in obstructive jaundice complicated with Gram-negative bacterial infection. (C) 2005 Blackwell Publishing Asia Pty Ltd.

Compensatory recovery of liver mass by Akt-mediated hepatocellular hypertrophy in liver-specific STAT3-deficient mice
S Haga, W Ogawa, H Inoue, K Terui, T Ogino, R Igarashi, K Takeda, S Akira, S Enosawa, H Furukawa, S Todo, M Ozaki
JOURNAL OF HEPATOLOGY, 43, 5, 799, 807, ELSEVIER SCIENCE BV, 2005年11月, ［査読有り］
英語, 研究論文（学術雑誌）, Background/Aims: Liver regeneration following hepatectomy is complicated and involves a variety of interacting factors. The present study was designed to study the roles of proliferation and hypertrophy of hepatocytes in liver regeneration following hepatectomy in liver-specific STAT3-knockout (LS3-KO) mice lacking mitogenic activity.
Methods: Partial hepatectomy was performed in LS3-KO and control mice. Liver regeneration was estimated by the liver weight, cell proliferation and cell size, and the related cellular signals were analyzed.
Results: Proliferation of hepatocytes following PH was markedly suppressed in LS3-KO mice with reduced cyclinD1 transcript. However, liver mass recovered sufficiently following PH in LS3-KO mice almost equal to that of control mice. Analysis of hepatocellular growth revealed that cell size following hepatectomy was significantly larger in LS3-KO mice than in control mice. Hepatectomy induced immediate but transient phosphorylation of Akt, p70(S6K), mTOR and GSK-3 beta in LS3-KO mice much more than in control mice. Additionally, adenoviral transfection of dominant negative mutant of Akt to control and LS3-KO mice led to insufficient liver regeneration following hepatectomy.
Conclusions: PI3-K/Akt-mediated responsive hepatocellular hypertrophy may be essential for liver regeneration following hepatectomy and sufficiently compensated liver regeneration even in STAT3-deficient liver, in which cell proliferation is impaired. (c) 2005 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Stat3 confers resistance against hypoxia/reoxygenation-induced oxidative injury in hepatocytes through upregulation of Mn-SOD
K Terui, S Enosawa, S Haga, HQ Zhang, H Kuroda, K Kouchi, T Matsunaga, H Yoshida, JF Engelhardt, K Irani, N Ohnuma, M Ozaki
JOURNAL OF HEPATOLOGY, 41, 6, 957, 965, ELSEVIER SCIENCE BV, 2004年12月, ［査読有り］
英語, 研究論文（学術雑誌）, Background/Aims: Hypoxia/reoxygenation (H/R) causes oxidative stress to the cell and induces apoptotic cell death. Signal transducer and activator of transcription-3 (Stat3) is one of the most important molecules involved in the initiation of liver development and regeneration, and has recently been shown to protect cells against various pathogens. In order to investigate the hepatoprotective effects of Stat3, we examined whether it protects against H/R-induced injury in primary hepatocytes.
Methods: Primary cultured hepatocytes were prepared from SD rats. Adenoviruses and cytokines were added 2 days and 1 h prior to the H/R insult, respectively. Hepatocytes and culture media were harvested for the assays before and after H/R insult.
Results: Interleukin-6 and cardiotropin-1, which may function mainly through Stat3 activation, protected cells from H/R-induced apoptosis. Adenoviral overexpression of the constitutively activated form of Stat3 (Stat3-C) reduced H/R-induced apoptosis as well as generation of reactive oxygen species (ROS) in hepatocytes. Interestingly, Stat3-C induced Mn-SOD, but not Cu/Zn-SOD, both at the protein and mRNA levels. Overexpression of Mn-SOD significantly reduced H/R-induced ROS and apoptosis by inhibiting redox-sensitive activation of caspase-3 activity.
Conclusions: Stat3 protects hepatocytes from H/R-induced cell injury at least partly by upregulating Mn-SOD and inactivating caspase-3. (C) 2004 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Improved hepatic regeneration with reduced injury by redox factor-1 in a rat small-sized liver transplant model
L Guo, S Haga, S Enosawa, K Naruse, Y Harihara, Y Sugawara, K Irani, M Makuuchi, M Ozaki
AMERICAN JOURNAL OF TRANSPLANTATION, 4, 6, 879, 887, BLACKWELL MUNKSGAARD, 2004年06月, ［査読有り］
英語, 研究論文（学術雑誌）, Redox factor-1 (Ref-1) has been shown to function in a redox-dependent manner in the cell. This study was designed to examine the effects of Ref-1 on liver regeneration as well as protection against postischemic injury in a rat model of 20% partial liver transplantation. Adenovirus carrying the full length of Ref-1 gene was introduced into liver grafts by ex vivo perfusion via the portal vein during preservation. Liver graft weights were assessed, as well as graft histology, serum levels of alanine aminotransferase (ALT)/bilirubin, DNA binding activities of AP-1 and Stat3. Redox factor-1 successfully expressed in the liver graft, improved regeneration by promoting cell proliferation. Overexpression of Ref-1 protein also reduced post-transplant injury and inflammatory reactions in the grafts. The increased serum levels of ALT and bilirubin observed after transplantation were significantly reduced by Ref-1 overexpression. Furthermore, adenovirally overexpressed Ref-1 in mouse liver successfully promoted liver regeneration after simple partial hepatectomy. Interestingly, Ref-1 significantly increased DNA binding of Stat3, but not AP-1. Overexpressed Ref-1 effectively promoted graft regeneration and reduced postischemic injury in a small-sized liver transplantation model. The results of the present study may open a new avenue to clinical transplantation of disproportionately sized grafts in living-related liver transplantation.

Hypoxia/re-oxygenation-induced, redox-dependent activation of STAT1 (signal transducer and activator of transcription 1) confers resistance to apoptotic cell death via hsp70 induction
K Terui, S Haga, S Enosawa, N Ohnuma, M Ozaki
BIOCHEMICAL JOURNAL, 380, Pt 1, 203, 209, PORTLAND PRESS, 2004年05月, ［査読有り］
英語, 研究論文（学術雑誌）, STAT1 (signal transducer and activator of transcription 1) is potentially involved in cell survival, as well as cell death, in different types of cells. The present study was designed to examine the effects of STAT1 on hypoxia/re-oxygenation (H/R)-induced cell death and/or survival, and the underlying mechanisms of any such effects. H/R was shown to induce apoptotic cell death of rat hepatocytes. The addition of a STAT1-specific inhibitor, fludarabine, significantly increased the fraction of apoptotic cells after H/R. Following H/R, STAT1 was activated and sequential phosphorylation of Tyr(701) and Ser(727) was observed, which could be inhibited by the antioxidant N-acetyl-L-cysteine. Tyrosine and serine phosphorylation of STAT1 was mediated by Janus kinase 2 and phosphoinositide 3-kinase/Akt kinase respectively in a redox-dependent manner following H/R. STAT I -induced HSP70 (heat-shock protein 70) expression and the suppression of apoptosis occurred concomitantly. In conclusion, STAT1 activation, in a redox-dependent manner, following H/R may play crucial roles in cell survival, at least partly via HSP70 induction.

Role of STAT-3 in regulation of hepatic gluconeogenic genes and carbohydrate metabolism in vivo
H Inoue, W Ogawa, M Ozaki, S Haga, M Matsumoto, K Furukawa, N Hashimoto, Y Kido, T Mori, H Sakaue, K Teshigawara, SY Jin, H Iguchi, R Hiramatsu, D LeRoith, K Takeda, S Akira, M Kasuga
NATURE MEDICINE, 10, 2, 168, 174, NATURE PUBLISHING GROUP, 2004年02月, ［査読有り］
英語, 研究論文（学術雑誌）, The transcription factor, signal transducer and activator of transcription-3 (STAT-3) contributes to various physiological processes. Here we show that mice with liver-specific deficiency in STAT-3, achieved using the Cre-loxP system, show insulin resistance associated with increased hepatic expression of gluconeogenic genes. Restoration of hepatic STAT-3 expression in these mice, using adenovirus-mediated gene transfer, corrected the metabolic abnormalities and the alterations in hepatic expression of gluconeogenic genes. Overexpression of STAT-3 in cultured hepatocytes inhibited gluconeogenic gene expression independently of peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha), an upstream regulator of gluconeogenic genes. Liver-specific expression of a constitutively active form of STAT-3, achieved by infection with an adenovirus vector, markedly reduced blood glucose, plasma insulin concentrations and hepatic gluconeogenic gene expression in diabetic mice. Hepatic STAT-3 signaling is thus essential for normal glucose homeostasis and may provide new therapeutic targets for diabetes mellitus.

Stat3 protects against Fas-induced liver injury by redox-dependent and -independent mechanisms
S Haga, K Terui, HQ Zhang, S Enosawa, W Ogawa, H Inoue, T Okuyama, K Takeda, S Akira, T Ogino, K Irani, M Ozaki
JOURNAL OF CLINICAL INVESTIGATION, 112, 7, 989, 998, AMER SOC CLINICAL INVESTIGATION INC, 2003年10月, ［査読有り］
英語, 研究論文（学術雑誌）, Signal transducer and activator of transcription-3 (Stat3) is one of the most important molecules involved in the initiation of liver development and regeneration. In order to investigate the hepatoprotective effects of Stat3, we examined whether Stat3 protects against Fas-mediated liver injury in the mouse. A constitutively activated form of Stat3 (Stat3-C) was adenovirally overexpressed in mouse liver by intravenous injection, and then a nonlethal dose of Fas agonist (Jo2) was injected intraperitoneally into the mouse (0.3 mug/g body wt). Stat3-C dramatically suppressed both apoptosis and necrosis induced by Jo2. In contrast, liver-specific Stat3-knockout mice failed to survive following Jo2 injection. Stat3-C upregulated expression of FLICE inhibitor protein (FLIP), Bcl-xL, and Bcl-2, and accordingly downregulated activities of FLICE and caspase-3 that were redox-independent. Interestingly, Stat3-C also upregulated the redox-associated protein redox factor-1 (Ref-1) and reduced apoptosis in liver following Jo2 injection by suppressing oxidative stress and redox-sensitive caspase-3 activity. These findings indicate that Stat3 activation protects against Fas-mediated liver injury by inhibiting caspase activities in redox-dependent and -independent mechanisms.

Inhibition of hypoxia/reoxygenation-induced oxidative stress in HGF-stimulated antiapoptotic siqnaling: role of P13-K and Akt kinase upon rac1
M Ozaki, S Haga, HQ Zhang, K Irani, S Suzuki
CELL DEATH AND DIFFERENTIATION, 10, 5, 508, 515, NATURE PUBLISHING GROUP, 2003年05月, ［査読有り］
英語, 研究論文（学術雑誌）, Rac1-regulated reactive oxygen species (ROS) production has been implicated in apoptosis. In contrast, pleiotropic protein kinase At protects against apoptosis. However, the pro- and antiapoptotic mechanisms of rac1 and Akt, respectively, and the intersection between these mechanisms are incompletely understood. In a model of oxidative stress and apoptosis induced by hypoxia/reoxygenation (H/R) in primary hepatocytes, activation of the PI3-K Akt axis by the prosurvival hepatocyte growth factor (HGF) inhibited H/R-stimulated rac1 activation and intracellular ROS production, and suppressed apoptosis. Suppression of PI3-K or Akt activity abrogated the inhibitory effect of HGF on rac1 activity and rac1-regulated oxidative stress. Furthermore, constitutive activation of Akt or PI3-K in the absence of HGF was sufficient to phosphorylate rac1, inhibit rac1 activation, and suppress rac1-regulated ROS production. These findings demonstrate that growth factor-stimulated activation of PI3-K-Akt is necessary and sufficient to suppress intracellular oxidative stress and apoptosis by inhibiting activation of proapoptotic, prooxidative rac1 GTPase.

Overexpression of redox factor-1 protects against postischemic liver injury by reducing oxidative stress and NF-kappa B activity .
M.Ozaki, S.Haga, K. Irani, H.Amemiya, S.Suzuki
Transplantation Proceedings, 34, 2640, 2642, 2002年, ［査読有り］

Regulatory action of toll-like receptor 2 in a non-alcoholic steatohepatitis mouse model
Min Yi, Masashi Kohanawa, Sanae Haga, Michitaka Ozaki, CYTOKINE, 100, 106, 106, 2017年12月
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD, 英語, 研究発表ペーパー・要旨（国際会議）

FUNCTION OF TOLL-LIKE RECEPTOR 2 IN THE MURINE INFLAMMATORY RESPONSE TO RHODOCOCCUS AURANTIACUS INFECTION
M. Yi, M. Kohanawa, S. Haga, M. Ozaki, CYTOKINE, 87, 131, 131, 2016年11月
ACADEMIC PRESS LTD- ELSEVIER SCIENCE LTD, 英語, 研究発表ペーパー・要旨（国際会議）

LC‐MS/MSを用いた血中チオ硫酸塩測定法の検討
神繁樹, 的場光太郎, 兵頭秀樹, 芳賀早苗, 尾崎倫孝, 質量分析総合討論会講演要旨集, 64th, 196, 2016年05月09日
日本語

PD-1-1 光を応用した移植細胞機能・細胞環境のモニタリングと制御の試み(PD-1 パネルディスカッション(1)細胞・臓器移植における基礎的研究の最前線,第114回日本外科学会定期学術集会)
尾崎 倫孝, 芳賀 早苗, 野田 なつみ, 森田 直樹, 山田 勇磨, 小澤 岳昌, 日本外科学会雑誌, 115, 2, 168, 168, 2014年03月05日
一般社団法人日本外科学会, 日本語

光学技術を応用した新たな臓器保存法，細胞･臓器移植法の開発
尾崎 倫孝, 芳賀 早苗, 森田 直樹, 野田 なつみ, 小澤 岳昌, Organ Biology, 21, 2, 171, 176, 2014年
In order to develop an effective organ/cell preservation method and to monitor/control post-transplant graft function continuously and non-invasively, an innovative optic technology to visualize/control cell/organ function seems to be useful. Recently, we have developed molecular probes to visualize redox states and cellular stresses, pH and cellular antigens in deeper lesions of the organ, and trying to activate intracellular key molecules by optical irradiation. In the hepatic ischemia/reperfusion model of mice, we successfully imaged liver oxidative stress and apoptosis by caspase-3 activity non-invasively and chronologically in a single mouse. We also developed a unique tool for visualizing intracellular pH and succeeded in imaging dynamic changes of pH in a mouse posterior limb ischemia/reperfusion model. Furthermore, we are now developing new tools to control intracellular molecules by blue light. We used plant-derived proteins to activate Akt/PKB molecule, essential for cell survival. By irradiating blue light, Akt in hepatocytes were phosphorylated(Ser and Thr)and activated. These tools will definitely provide a new avenue toward cell/organ transplantation in the future., The Japan Society for Organ Preservation and Biology, 日本語

A Method for Intravital Monitoring of Human Cells Using a Far-Red Luminescent Probe in Graft-Versus-Host Disease Model Mice (vol 133, pg 841, 2013)
Hongjiang Qiao, Riichiro Abe, Nao Saito, Yasuyuki Fujita, Inkin Hayashi-Ujiie, Gang Wang, Sanae Haga, Chun Wu, Yoshihiro Ohmiya, Michitaka Ozaki, Hiroshi Shimizu, JOURNAL OF INVESTIGATIVE DERMATOLOGY, 133, 6, 1692, 1692, 2013年06月
NATURE PUBLISHING GROUP, 英語, その他

Bioluminescent imaging of hepatic caspase-3 activity in mice
Michitaka Ozaki, Sanae Haga, Takeaki Ozawa, FASEB JOURNAL, 27, 2013年04月
FEDERATION AMER SOC EXP BIOL, 英語, 研究発表ペーパー・要旨（国際会議）

PD-4-1 肝再生における分子メカニズムの解析(PD パネルディスカッション,第113回日本外科学会定期学術集会)
尾崎 倫孝, 芳賀 早苗, 井上 啓, 小川 渉, 小澤 岳彗, 日本外科学会雑誌, 114, 2, 236, 236, 2013年03月05日
一般社団法人日本外科学会, 日本語

カスパーゼ3活性化プローブによる肝傷害の生体イメージング
尾崎 倫孝, 芳賀 早苗, 小澤 岳昌, 北海道醫學雜誌 = Acta medica Hokkaidonensia, 87, 6, 253, 253, 2012年11月01日
日本語

STAT3 plays a pivotal role in cell adhesion in mouse hepatocytes by regulating E-cadherin expression
S. Haga, H. Inoue, A. Kano, X. -Y. Fu, K. Terui, M. Ozaki, VIRCHOWS ARCHIV, 461, S166, S167, 2012年09月
SPRINGER, 英語, 研究発表ペーパー・要旨（国際会議）

生体イメージング法による細胞・臓器のリアルタイム評価法の開発
芳賀 早苗, 尾崎 倫孝, Organ Biology, 19, 1, 29, 35, 2012年
The organ function and its regulatory mechanisms which are affected by various physiopathological conditions have been analyzed by molecular and biochemical methods. We have been working on developing various optical probes to visualize cellular and organ conditions/functions in vivo. An innovative technology is essential for imaging the biological functions, especially by using bioluminescence；for example, a novel highly transmittable and sensitive optic probe. We have, so far, developed some novel probes based on the unique bioluminescence theories. By applying these probes for biological systems, we succeeded to visualize the dynamic changes of liver injury and oxidative stress in mouse liver ischemia/reperfusion model by the caspase-3 activity probe and redox sensitive GFP probe, respectively. These novel bio-imaging techniques will definitely bring great possibility and potential impact for research and clinical medicine in the future., The Japan Society for Organ Preservation and Biology, 日本語

SF-003-5 新規臓器保存液(FJ液)による肝冷保存再灌流障害の軽減 : FJ液の重要な成分(SF-003 サージカルフォーラム(3)移植-2,第111回日本外科学会定期学術集会)
旭 火華, 深井 原, 福森 大介, 若山 顕治, 山下 健一郎, 廣方 玄太郎, 芳賀 早苗, 田原 宗徳, 谷口 雅彦, 嶋村 剛, 鈴木 友己, 松下 通明, 古川 博之, 尾崎 倫孝, 藤堂 省, 日本外科学会雑誌, 112, 1, 340, 340, 2011年05月25日
一般社団法人日本外科学会, 日本語

PS-071-5 新規薬剤DTCM-Gの免疫抑制効果に関する検討(PS-071 ポスターセッション(71)移植,第111回日本外科学会定期学術集会)
柴崎 晋, 山下 健一郎, 後藤 了一, 若山 顕治, 常俊 雄介, 財津 雅昭, 五十嵐 瑠美, 芳賀 早苗, 尾崎 倫孝, 梅澤 一夫, 藤堂 省, 日本外科学会雑誌, 112, 1, 627, 627, 2011年05月25日
一般社団法人日本外科学会, 日本語

PS-071-7 低温下で好気代謝を促進する新規臓器保存液の至適条件 : 低温酸素化灌流保存法への応用を目指した基礎的検討(PS-071 ポスターセッション(71)移植,第111回日本外科学会定期学術集会)
深井 原, 若山 顕治, 山下 健一郎, 福森 大介, 廣方 玄太郎, 芳賀 早苗, 谷口 雅彦, 嶋村 剛, 鈴木 友己, 尾崎 倫孝, 松下 通明, 古川 博之, 藤堂 省, 日本外科学会雑誌, 112, 1, 628, 628, 2011年05月25日
一般社団法人日本外科学会, 日本語

A Novel Immunosuppressant, DTCM-G, Inhibits T-Cells Via p70 S6K and Prolongs Cardiac Allograft Survival in Mice.
S. Shibasaki, K. Yamashita, R. Goto, K. Wakayama, Y. Tsunetoshi, M. Zaitsu, S. Haga, R. Igarashi, M. Ozaki, K. Umezawa, S. Todo, AMERICAN JOURNAL OF TRANSPLANTATION, 11, 474, 474, 2011年04月
WILEY-BLACKWELL, 英語, 研究発表ペーパー・要旨（国際会議）

移植臓器の非侵襲的モニタリング法の開発
尾崎 倫孝, 芳賀 早苗, 森田 直樹, 深井 原, 藤堂 省, 小澤 岳昌, 近江谷 克裕, Organ Biology, 18, 1, 134, 140, 2011年
In order to develop an effective organ/cell preservation solution and to monitor graft function continuously and non-invasively, an innovative imaging technology to visualize cell/organ function is required. Recently, we developed molecular probes to visualize redox states and cellular stresses,and cellular antigens in deeper lesions of the organ. In the hepatic ischemia/reperfusion model of mice, we successfully imaged liver oxidative stress and apoptosis(by caspase-3 activity)noninvasively and chronologically in a single mouse. We are also trying to develop new tools to visualize ER stress and cellular antigens in deeper lesions. These tools will definitely provide a new avenue toward cell/organ transplantation in the future., The Japan Society for Organ Preservation and Biology, 日本語

新規化合物NK026680を用いたex‐vivo誘導制御性樹状細胞の免疫抑制効果
柴崎晋, 山下健一郎, 若山顕治, 廣方玄太郎, 常俊雄介, 財津雅昭, 五十嵐瑠美, 芳賀早苗, 尾崎倫孝, 藤堂省, 移植, 45, 179, 2010年10月
日本語

A Novel Solution for Rat Liver Preservation Composed of Heavy Water and Swan Buffer; a Pilot Study
Daisuke Fukumori, Moto Fukai, Kenji Wakayama, Kenichiro Yamashita, Shinya Ueki, Mitsuru Sugawara, Sanae Haga, Hiroyuki Furukawa, Michitaka Ozaki, Satoru Todo, AMERICAN JOURNAL OF TRANSPLANTATION, 10, 496, 497, 2010年04月
WILEY-BLACKWELL PUBLISHING, INC, 英語, 研究発表ペーパー・要旨（国際会議）

Heavy Water (D2O) Universally Ameliorates Cold Preservation Injury: Precise Analyses of Cytoprotective Mechanisms In Vitro
Moto Fukai, Kenji Wakayama, Daisuke Fukumori, Kenichiro Yamashita, Mitsuru Sugawara, Sanae Haga, Ueki Gentaro Hirokata, Hiroyuki Furukawa, Michitaka Ozaki, Satoru Todo, AMERICAN JOURNAL OF TRANSPLANTATION, 10, 492, 493, 2010年04月
WILEY-BLACKWELL, 英語, 研究発表ペーパー・要旨（国際会議）

コラーゲンビトリゲル薄膜の特徴を活用した新しい細胞培養システムの開発
竹澤 俊明, 福田 真野, Mcintosh-Ambrose Winnette, 高 知愛, Elisseeff Jennifer, 芳賀 早苗, 尾崎 倫孝, 加藤 聖子, 王 碧昭, 内野 正, 西田 輝夫, 薬学雑誌, 130, 4, 565, 574, 2010年04月
高密度のコラーゲン線維のゲルをガラス化工程を経て作製したコラーゲンビトリゲル(CV)は、薄膜状、糸状、管状など様々な形状に整形できる。このCV薄膜の特徴について述べ、これを用いた以下の細胞培養システムや薬物送達システムの開発を紹介した。1)眼刺激性および透過性試験用の角膜細胞培養システム、2)皮膚感作性試験用の皮膚細胞培養システム、3)血漿成分の濾過モデル用の腎糸球体細胞培養システム、4)肝障害を改善するための薬物送達システム。更に、このCVを創薬支援ツールとして展開することの有用性について述べた。, (公社)日本薬学会, 日本語

WS-6-6 胃癌腹膜播種に対する,NF-κB抑制による治療戦略 : 腹膜との接着抑制を主眼に置いて(消化器癌腹膜転移の治療:基礎と臨床,ワークショップ(6),第110回日本外科学会定期学術集会)
三野 和宏, 中西 一彰, 芳賀 早苗, 馬場 基, 藤好 真人, 佐藤 正法, 横尾 英樹, 神山 俊哉, 尾崎 倫孝, 梅澤 一夫, 藤堂 省, 日本外科学会雑誌, 111, 2, 166, 166, 2010年03月05日
一般社団法人日本外科学会, 日本語

WS-6-10 NF-κB制御による膵癌腹膜播種の抑制 : アノイキス誘導による新たな治療戦略(消化器癌腹膜転移の治療:基礎と臨床,ワークショップ(6),第110回日本外科学会定期学術集会)
佐藤 正法, 中西 一彰, 渡邉 俊之, 芳賀 早苗, 三野 和宏, 横尾 英樹, 梅澤 一夫, 近江谷 克裕, 神山 俊哉, 尾崎 倫孝, 藤堂 省, 日本外科学会雑誌, 111, 2, 167, 167, 2010年03月05日
一般社団法人日本外科学会, 日本語

OP-018-4 Magnet cell sorting(MACS) systemを用いた大腸癌の血中腫瘍細胞の同定(腫瘍マーカー-1,一般口演,第110回日本外科学会定期学術集会)
崎浜 秀康, 小林 希, 佐々木 彩実, 芳賀 早苗, 本間 重紀, 高橋 典彦, 片岡 昭彦, 尾崎 倫孝, 藤堂 省, 日本外科学会雑誌, 111, 2, 323, 323, 2010年03月05日
一般社団法人日本外科学会, 日本語

OP-119-2 NK026680を用いたRegulatory DCによる免疫抑制効果の検討(移植-1,一般口演,第110回日本外科学会定期学術集会)
柴崎 晋, 山下 健一郎, 若山 顕治, 廣方 玄太郎, 常俊 雄介, 財津 雅昭, 木村 鐘康, 五十嵐 瑠美, 芳賀 早苗, 谷口 雅彦, 鈴木 友己, 嶋村 剛, 古川 博之, 尾崎 倫孝, 藤堂 省, 日本外科学会雑誌, 111, 2, 477, 477, 2010年03月05日
一般社団法人日本外科学会, 日本語

OP-152-2 重水含有新規臓器保存液の開発 : 効果の普遍性と保護作用メカニズムの検討(in vitro study)(移植-8,一般口演,第110回日本外科学会定期学術集会)
深井 原, 若山 顕治, 福森 大介, 山下 健一郎, 後藤 了一, 植木 伸也, 柴崎 晋, 廣方 玄太郎, 木村 鐘康, 常俊 雄介, 財津 雅明, 芳賀 早苗, 谷口 雅彦, 鈴木 友己, 嶋村 剛, 古川 博之, 松下 通明, 尾崎 倫孝, 藤堂 省, 日本外科学会雑誌, 111, 2, 525, 525, 2010年03月05日
一般社団法人日本外科学会, 日本語

生体イメージングをもちいたHGF含有コラーゲンビトリゲル薄膜による肝虚血/再灌流障害抑制効果の検討
芳賀早苗, 竹澤俊明, 山本智子, 小澤岳昌, 森田直樹, 藤好真人, 尾崎倫孝, 再生医療, 9, 304, 2010年02月05日
日本語

p66Shc Plays a Pivotal Role in Impaired Liver Regeneration in Aged Mice by a Redox-dependent Mechanism
Michitaka Ozaki, S. Haga, N. Morita, T. Ozawa, S. Todo, K. Irani, J. Remington, FREE RADICAL BIOLOGY AND MEDICINE, 49, S83, S83, 2010年
ELSEVIER SCIENCE INC, 英語, 研究発表ペーパー・要旨（国際会議）

コラーゲンビトリゲル薄膜の特徴を活用した新しい細胞培養システムの開発
竹澤 俊明, 福田 真野, Mcintosh-Ambrose Winnette, 高 知愛, ELISSEEFF Jennifer, 芳賀 早苗, 尾崎 倫孝, 加藤 聖子, 王 碧昭, 内野 正, 西田 輝夫, 薬学雑誌. 乙号, 130, 4, 565, 574, 2010年
The white of an egg, rendered opaque by boiling, can be converted into a thin, transparent and rigid material like glass by evaporating the moisture. This phenomenon is known as the vitrification of heat-denatured proteins. We applied vitrification technology to a collagen gel and converted it into a rigid glass-like material. We attempted to rehydrate the glass-like material and succeeded in preparing a novel stable state of collagen gel that was a thin and transparent membrane with excellent gel strength and protein permeability. We called it "collagen vitrigel" because it was produced from the vitrification process of a traditional hydrogel. Further, a framework-embedded collagen vitrigel membrane that can be easily turned inside out with tweezers was prepared by inserting a nylon membrane ring in the collagen sol prior to the gelation, thereby allowing the membrane to function as a removable cell culture substratum. Different types of anchorage-dependent cells could be cultured on both surfaces of the substratum by the manipulation of two-dimensional cultures, and consequently a three-dimensional crosstalk model with paracrine effects from each cell type was reconstructed. Also, the collagen vitrigel membrane containing a bioactive molecule provided a drug delivery system (DDS) with sustainable release. In this review, we summarize the recent progress of applied studies using the collagen vitrigel membrane as follows: a corneal model for eye irritant and permeability tests, a skin model for sensitization test, a renal glomerular model for evaluating blood filtration, an endometrial model for developing a new treatment and a DDS of hepatocyte growth factor for improving liver disorder., The Pharmaceutical Society of Japan, 日本語

NK026680のT細胞抑制効果ならびにラット心移植における免疫抑制効果
柴崎晋, 山下健一郎, 若山顕治, 廣方玄太郎, 木村鐘康, 財津雅昭, 常俊雄介, 五十嵐瑠美, 芳賀早苗, 藤堂省, 移植, 44, 259, 2009年09月
日本語

組織再生に有用なコラーゲンビトリゲルを用いた新しい医薬学研究:肝虚血・再灌流後の肝障害抑制への応用
竹澤俊明, 芳賀早苗, 尾崎倫孝, Pharmaco-Hematologyシンポジウム講演要旨集, 10th, 25, 2009年06月19日
日本語

新しい細胞培養システムの開発とその創薬支援研究への応用 コラーゲンビトリゲル薄膜の特徴を活用した新しい細胞培養システムの開発　　　　　　　　　　　　　　　
竹澤 俊明, 福田 真野, Ambrose Winnette Mcintosh, 高 知愛, Elisseeff Jennifer H, 芳賀 早苗, 尾崎 倫孝, 加藤 聖子, 王 碧昭, 内野 正, 西田 輝夫, 日本薬学会年会要旨集, 129年会, 1, 179, 179, 2009年03月
(公社)日本薬学会, 日本語

SF-111-2 マウス胃癌腹膜播種モデルを用いた,新規NFκB阻害剤DHMEQによる腫瘍抑制効果の検討(分子標的・遺伝子治療1,サージカルフォーラム,第109回日本外科学会定期学術集会)
三野 和宏, 中西 一彰, 芳賀 早苗, 喜納 政哉, 佐藤 正法, 横尾 英樹, 神山 俊哉, 尾崎 倫孝, 藤堂 省, 梅澤 一夫, 日本外科学会雑誌, 110, 2, 363, 363, 2009年02月25日
一般社団法人日本外科学会, 日本語

HP-189-4 Suboptimal graftの有効利用を目指した次世代の臓器保存液の開発(移植医療2,ハイブリッドポスター,第109回日本外科学会定期学術集会)
深井 原, 福森 大介, 若山 顕治, 山下 健一郎, 後藤 了一, 植木 伸也, 柴崎 晋, 大浦 哲, 廣方 玄太郎, 芳賀 早苗, 谷口 雅彦, 鈴木 友己, 嶋村 剛, 松下 通明, 古川 博之, 尾崎 倫孝, 藤堂 省, 日本外科学会雑誌, 110, 2, 710, 710, 2009年02月25日
一般社団法人日本外科学会, 日本語

HP-090-6 胃癌腹膜播種モデルにおける新規NFκB阻害剤の腹膜への細胞接着抑制効果の検討(胃(分子標的・免疫療法),ハイブリッドポスター,第109回日本外科学会定期学術集会)
佐藤 正法, 三野 和宏, 中西 一彰, 芳賀 早苗, 上徳 ひろみ, 喜納 政哉, 横尾 英樹, 神山 俊哉, 梅澤 一夫, 尾崎 倫孝, 藤堂 省, 日本外科学会雑誌, 110, 2, 549, 549, 2009年02月25日
一般社団法人日本外科学会, 日本語

新規開発臓器保存液を用いた心冷保存限界延長の試み
若山 顕治, 深井 原, 山下 健一郎, 後藤 了一, 植木 伸也, 福森 大介, 柴崎 晋, 大浦 哲, 廣方 玄太郎, 芳賀 早苗, 谷口 雅彦, 鈴木 友己, 嶋村 剛, 松下 道明, 古川 博之, 尾崎 倫孝, 藤堂 省, 小野 太祐, 絹川 真太郎, 筒井 裕之, 菅原 満, 日本外科学会雑誌, 110, 臨増2, 709, 709, 2009年02月
(一社)日本外科学会, 日本語

新規開発臓器保存液を用いた心冷保存限界延長の試み
若山 顕治, 深井 原, 山下 健一郎, 後藤 了一, 植木 伸也, 福森 大介, 柴崎 晋, 大浦 哲, 廣方 玄太郎, 芳賀 早苗, 谷口 雅彦, 鈴木 友己, 嶋村 剛, 松下 道明, 古川 博之, 尾崎 倫孝, 藤堂 省, 小野 太祐, 絹川 真太郎, 筒井 裕之, 菅原 満, 日本外科学会雑誌, 110, 臨増2, 709, 709, 2009年02月
(一社)日本外科学会, 日本語

A Novel Triazolopyrimidine Derivative, NK026680 Inhibits T Cell Function, and Prolongs Cardiac Allograft Survival in Rats.
Susumu Shibasaki, Kenichiro Yamashita, Ryoichi Goto, Tetsu Oura, Kenji Wakayama, Gentaro Hirokata, Tomohiro Shibata, Rumi Igarashi, Sanae Haga, Masahiko Taniguchi, Tomomi Suzuki, Tsuyoshi Shimamura, Hiroyuki Furukawa, Michitaka Ozaki, Satoru Todo, AMERICAN JOURNAL OF TRANSPLANTATION, 9, 660, 660, 2009年
WILEY-BLACKWELL PUBLISHING, INC, 英語, 研究発表ペーパー・要旨（国際会議）

IMPORTANCE OF STAT3 AND PDK1/AKT PATHWAYS IN LIVER REGENERATION IN A MOUSE HEPATECTOMY MODEL: ROLES OF CELL PROLIFERATION AND SIZE
M. Ozaki, S. Haga, H. Inoue, Y. Okamoto, W. Ogawa, K. Terui, N. Morita, K. Takeda, S. Akira, S. Todo, JOURNAL OF HEPATOLOGY, 50, S312, S312, 2009年
ELSEVIER SCIENCE BV, 英語, 研究発表ペーパー・要旨（国際会議）

Re-Evaluation of Heavy Water for Organ Preservation Solution; In Vitro Study.
Moto Fukai, Kenichiro Yamashita, Kenji Wakayama, Daisuke Fukumori, Ryoichi Goto, Sanae Haga, Gentaro Hirokata, Shigeru Nakakimura, Mitsuru Sugawara, Hirofumi Kamachi, Tomomi Suzuki, Tsuyoshi Shimamura, Hiroyuki Furukawa, Michiaki Matsushita, Michitaka Ozaki, Satoru Todo, AMERICAN JOURNAL OF TRANSPLANTATION, 9, 581, 581, 2009年
WILEY-BLACKWELL PUBLISHING, INC, 英語, 研究発表ペーパー・要旨（国際会議）

NON-INVASIVE REALTIME MONITORING OF OXIDATIVE STRESS AND DAMAGE BY NEWLY DEVELOPED OPTICAL IMAGING PROBES IN MOUSE LIVER ISCHEMIA/REPERFUSION
Michitaka Ozaki, Sanae Haga, S. James Remington, Takeaki Ozawa, Yuko Oikawa, Satoru Todo, HEPATOLOGY, 48, 4, 640A, 640A, 2008年10月
JOHN WILEY & SONS INC, 英語, 研究発表ペーパー・要旨（国際会議）

A novel NF-kappa B inhibitor, DHMEQ, attenuates arterial vasculopathy.
Motohiro Uno, Kenichiro Yamashita, Tomohiro Shibata, Takeshi Aoyagi, Tetu Oura, Sanae Haga, Masaaki Watanabe, Kazuo Umezawa, Michitaka Ozaki, Satoru Todo, AMERICAN JOURNAL OF TRANSPLANTATION, 8, 343, 344, 2008年05月
BLACKWELL PUBLISHING, 英語, 研究発表ペーパー・要旨（国際会議）

Induction of immunosuppressive dendritic cells by a novel NF-kappa B inhibitor, DHMEQ
Takeshi Aoyagi, Kenichiro Yamashita, Yoshiki Yanagawa, Ryoichi Goto, Rumi Igarashi, Sanae Haga, Michitaka Ozaki, Kazunori Onoe, Kazuo Umezawa, Hiroyuki Furukawa, Satoru Todo, AMERICAN JOURNAL OF TRANSPLANTATION, 8, 483, 483, 2008年05月
BLACKWELL PUBLISHING, 英語, 研究発表ペーパー・要旨（国際会議）

肝移植における虚血・再灌流障害のシグナル伝達機構
深井 原, 芳賀 早苗, 尾崎 倫孝, 臨床化学, 37, 2, 131, 140, 2008年04月30日
日本語

SF-018-2 新規NFκB阻害剤DHMEQの動脈硬化抑制効果(第108回日本外科学会定期学術集会)
廣方 玄太郎, 宇野 元博, 山下 健一郎, 柴田 智裕, 後藤 了一, 青柳 武史, 大浦 哲, 渡辺 正明, 芳賀 早苗, 谷口 雅彦, 鈴木 友己, 嶋村 剛, 古川 博之, 梅澤 一夫, 尾崎 倫孝, 藤堂 省, 日本外科学会雑誌, 109, 2, 208, 208, 2008年04月25日
一般社団法人日本外科学会, 日本語

SF-049-2 新規NF-κB阻害剤DHMEQの各種免疫担当細胞に対する効果(第108回日本外科学会定期学術集会)
大浦 哲, 山下 健一郎, 青柳 武史, 松本 秀一郎, 石戸谷 裕樹, 柳川 芳毅, 後藤 了一, 宇野元 博, 五十嵐 瑠美, 芳賀 早苗, 渡辺 正明, 柴田 智裕, 谷口 雅彦, 鈴木 友己, 嶋村 剛, 古川 博之, 小野江 和則, 尾崎 倫孝, 梅澤 一夫, 藤堂 省, 日本外科学会雑誌, 109, 2, 247, 247, 2008年04月25日
一般社団法人日本外科学会, 日本語

DP-036-3 新規NF-kB阻害剤Dehydroxymethylepoxyquinomicin (DHMEQ)を用いた肝温阻血再灌流傷害軽減の試み(第108回日本外科学会定期学術集会)
深井 原, 尾崎 倫孝, 芳賀 早苗, 山下 健一郎, 谷口 雅彦, 鈴木 友己, 嶋村 剛, 松下 通明, 古川 博之, 岸野 吏志, 梅澤 一夫, 藤堂 省, 日本外科学会雑誌, 109, 2, 416, 416, 2008年04月25日
一般社団法人日本外科学会, 日本語

DP-121-4 生体イメージングに向けて新規開発した分子機能プローブの基礎的検討(第108回日本外科学会定期学術集会)
尾崎 倫孝, 芳賀 早苗, 及川 優子, 小澤 岳昌, 古川 博之, 藤堂 省, 日本外科学会雑誌, 109, 2, 586, 586, 2008年04月25日
一般社団法人日本外科学会, 日本語

肝移植における虚血・再灌流障害のシグナル伝達機構
深井 原, 芳賀 早苗, 尾崎 倫孝, 臨床化学, 37, 2, 131, 140, 2008年
Japan Society of Clinical Chemistry, 日本語

バイオイメージングに向けた光プローブの基礎的研究
芳賀 早苗, 及川 優子, 小澤 岳昌, 藤堂 省, 尾崎 倫孝, バイオイメージング, 16, 2, 161, 162, 2007年10月05日
日本語

P66SHC, an ageing protein, plays a pivotal role in post-hepatectomized liver regeneration in aged mice
Michitaka Ozaki, Sanae Haga, Tadahiro Osada, Yuko Oikawa, Hiroyuki Furukawa, Kaikobad Irani, Satoru Todo, HEPATOLOGY, 46, 4, 536A, 537A, 2007年10月
JOHN WILEY & SONS INC, 英語, 研究発表ペーパー・要旨（国際会議）

WS-11-5 種々の病態における肝再生不全の要因 : 細胞内シグナル解析からみた問題点と対策(第107回日本外科学会定期学術集会)
尾崎 倫孝, 芳賀 早苗, 長田 忠大, 井上 啓, 小川 渉, 村田 宏, 岩垣 博巳, 古川 博之, 藤堂 省, 日本外科学会雑誌, 108, 2, 159, 159, 2007年03月10日
一般社団法人日本外科学会, 日本語

Prevention of liver ischemia/reperfusion-induced injury : from bench to clinics
尾崎 倫孝, 芳賀 早苗, 深井 原, 古川 博之, 梅澤 一夫, 藤堂 省, 日本外科学会雑誌, 107, 2, 84, 84, 2006年03月05日
一般社団法人日本外科学会, 英語

Ex vivo adenoviral gene transfer of constitutively activated STAT3 reduced ischemia-reperfusion induced injury and promoted liver regeneration in 20% rat partial liver transplant model.
M Ozaki, S Haga, L Guo, H Furukawa, M Matsushita, S Todo, LIVER TRANSPLANTATION, 11, 7, C34, C34, 2005年07月
JOHN WILEY & SONS INC, 英語, 研究発表ペーパー・要旨（国際会議）

A-15 STAT3の細胞保護作用を用いた再生医療への応用 : 低酸素再酸素化モデルにおける検討(小児外科としての発生学と再生医学)
照井 慶太, 尾崎 倫孝, 芳賀 早苗, 山田 慎一, 菱木 知郎, 黒田 浩明, 幸地 克憲, 松永 正訓, 吉田 英生, 大沼 直躬, 日本小児外科学会雑誌, 41, 3, 372, 372, 2005年05月20日
特定非営利活動法人日本小児外科学会, 日本語

肝虚血再灌流傷害におけるSTAT3の細胞保護作用(第105回日本外科学会定期学術集会)
照井 慶太, 尾崎 倫孝, 芳賀 早苗, 絵野 沢伸, 黒田 浩明, 幸地 克憲, 松永 正訓, 吉田 英生, 大沼 直躬, 日本外科学会雑誌, 106, 0, 419, 419, 2005年04月05日
一般社団法人日本外科学会, 日本語

Constitutive activation of STAT3 confers resistance to liver against Fas-mediated liver injury: Redox-dependent and -independent mechanisms.
M Ozaki, S Haga, W Ogawa, H Inoue, K Terui, S Enosawa, HEPATOLOGY, 40, 4, 566A, 566A, 2004年10月
JOHN WILEY & SONS INC, 英語, 研究発表ペーパー・要旨（国際会議）

B-27 肝虚血再灌流傷害におけるSTAT3の細胞保護作用(一般口演 移植I)
照井 慶太, 尾崎 倫孝, 芳賀 早苗, 絵野沢 伸, 吉田 英生, 松永 正訓, 幸地 克憲, 黒田 浩明, 斎藤 武, 大沼 直躬, 日本小児外科学会雑誌, 40, 3, 420, 420, 2004年05月20日
特定非営利活動法人日本小児外科学会, 日本語

活性化Stat3(S3‐C)による劇症肝炎の抑制機序と治療への可能性
尾崎倫孝, 芳賀早苗, 照井慶太, 小川渉, 井上啓, 絵野沢伸, 肝臓, 45, Supplement 1, A75, 2004年04月30日
日本語

SF-093-1 肝再生おける細胞内シグナルの解析 : Stat3経路およびPI3-K/PDK/Akt経路の重要性
芳賀 早苗, 照井 慶太, 張 慧〓, 小川 渉, 井上 啓, 絵野沢 伸, 尾崎 倫孝, 日本外科学会雑誌, 105, 0, 299, 299, 2004年03月15日
一般社団法人日本外科学会, 日本語

Analysis of regulatory signals of liver regeneration - Signals regulating hepatocellular proliferation and growth.
S Haga, S Enosawa, K Terui, W Ogawa, H Inoue, M Ozaki, AMERICAN JOURNAL OF TRANSPLANTATION, 4, 465, 465, 2004年
BLACKWELL MUNKSGAARD, 英語, 研究発表ペーパー・要旨（国際会議）

Stat3 confers resistance against hypoxia/reoxygenation-induced oxidative injury in hepatocytes via Mn-SOD.
K Terui, S Haga, S Enosawa, N Onuma, M Ozaki, AMERICAN JOURNAL OF TRANSPLANTATION, 4, 392, 392, 2004年
BLACKWELL MUNKSGAARD, 英語, 研究発表ペーパー・要旨（国際会議）

肝虚血再潅流傷害におけるSTAT3の細胞保護作用
照井慶太, 尾崎倫孝, 芳賀早苗, 張慧ご, 菅沼靖之, 大沼直み, 絵野沢伸, 移植, 38, 247, 2003年10月
日本語

Constitutive activation of Stat3 by adenoviral gene transfer confers resistance to liver against Fas-mediated liver injury: Caspase inhibition and redox-dependent mechanisms
M Ozaki, S Haga, HQ Zhang, K Terui, W Ogawa, H Inoue, T Okuyama, K Takeda, S Akira, S Enosawa, S Suzuki, MOLECULAR THERAPY, 7, 5, S59, S59, 2003年05月
ACADEMIC PRESS INC ELSEVIER SCIENCE, 英語, 研究発表ペーパー・要旨（国際会議）

A-024 STAT3 の細胞保護作用を用いた再生医療への応用 : 肝の低酸素再酸素化モデルにおける検討
照井 慶太, 尾崎 倫孝, 芳賀 早苗, 張 慧き, 菅沼 靖之, 絵野沢 伸, 鈴木 盛一, 大沼 直躬, 日小外会誌, 39, 3, 106, 106, 2003年
特定非営利活動法人 日本小児外科学会, 日本語

低酸素再酸素化によるSTAT1のレドックス制御と肝細胞生存への影響
照井慶太, 尾崎倫孝, 芳賀早苗, 絵野沢伸, 鈴木盛一, 移植, 37, 269, 2002年09月
日本語

HGFによる肝虚血再潅流傷害抑制メカニズム : PI3-K/Akt/Racl pathwayによる細胞内活性酸素消去系
芳賀 早苗, 尾崎 倫孝, 長田 夕子, 鈴木 盛一, 日本外科学会雑誌, 103, 252, 252, 2002年03月10日
一般社団法人日本外科学会, 日本語

シグナル制御による臓器虚血再潅流傷害抑制の試み
尾崎 倫孝, 田中 秀明, 郭 雷, 芳賀 早苗, Irani Kaikobad, 鈴木 盛一, 日本外科学会雑誌, 103, 197, 197, 2002年03月10日
一般社団法人日本外科学会, 日本語

光プローブをもちいた肝の酸化ストレスと傷害（プログラム細胞死）の観察と評価　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 森田 直樹, 小澤 岳昌
第28回肝細胞研究会, 2021年09月10日, シンポジウム・ワークショップパネル（公募）
2021年09月10日 - 2021年09月11日

肝虚血再灌流傷害進展における Poly（ADP-ribose）polymerase（PARP）依存性細胞死の解析　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 浅野 真未, 菅野 憲, 小澤 岳昌, 森田 直樹
第27回肝細胞研究会, 2020年12月15日, 口頭発表（一般）
2020年12月16日

様々な様式の細胞死は、慢性脂肪肝における遷延化した組織傷害に関与する可能性がある　　　　　　　　　　　　　　　
芳賀 早苗, 森田 直樹, 小澤 岳昌, 尾崎 倫孝
第93回日本生化学会大会, ポスター発表
2020年09月14日 - 2020年09月16日

Fasリガンド/酸化ストレスによって引き起こされるプログラム細胞死の機序解析 Analysis of programmed cell death in hepatocytes induced by Fas ligand and oxidative stress　　　　　　　　　　　　　　　
芳賀早苗, 森田直樹, 尾崎倫孝
第92回日本生化学会総会, 2019年09月20日, 日本語, ポスター発表
［国内会議］

脂肪肝における易傷害性メカニズム解析の基礎的研究　　　　　　　　　　　　　　　
芳賀早苗, 浅野真未, 森田直樹, 尾崎倫孝
第26回肝細胞研究会, 2019年05月24日, 日本語, ポスター発表
［国内会議］

Relevance of FXR-p62/SQSTM1 pathway for survival and protection of mouse hepatocytes and liver with steatosis.　　　　　　　　　　　　　　　
Michitaka Ozaki, Sanae Haga, Yimi
IBD and Liver: East Meets West, 2018年
京都ホテルオークラ、京都

「発光プローブによるプログラム細胞死（アポトーシス、ネクロプトーシス）動的解析の試み」　　　　　　　　　　　　　　　
芳賀 早苗, 菅野 憲, 小澤 岳昌, 森田 直樹, 浅野 真未, 伊 敏, 尾崎 倫孝
第27回日本Cell Death学会, 2018年
京都府立医科大学 図書館ホール、京都

「光による細胞生存能（Akt/PKB分子）制御に関する研究」　　　　　　　　　　　　　　　
芳賀 早苗, 小澤 岳昌, 森田 直樹, 伊 敏, 尾崎 倫孝
第91回日本生化学会, 2018年
国立京都国際会館、京都

「マウス脂肪肝モデルをもちいたビルベリーの予防効果に関する検討」　　　　　　　　　　　　　　　
芳賀 早苗, 伊 敏, 森田 直樹, 浅野 真未, 荘厳 哲哉, 尾崎 倫孝
第39会日本肥満学会, 2018年
神戸国際会議場、神戸

「レドックスが制御するネクローシス型細胞死（ネクロプトーシス）の光プローブによる動的解析」　　　　　　　　　　　　　　　
芳賀 早苗, 菅野 憲, 小澤 岳昌, 森田 直樹, 浅野 真未, 尾崎 倫孝
第25回肝細胞研究会, 2018年
東京大学伊藤謝恩ホール、東京

「光プローブをもちいたプログラム細胞死（アポトーシス、ネクロプトーシス）の動的解析」　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗
第22回日本がん分子標的治療学会学術集会, 2018年
都市センターホテル、東京

Spatio-temporal imaging of cell/organ physio-pathology and its clinical application.　　　　　　　　　　　　　　　
Michitaka Ozaki, Sanae Haga, Toshiyuki Hamada, Takeaki Ozawa, Yuma Yamada
CIS Workshop 2017 “Innovative Bio-imaging toward Diagnosis and Therapy”, 2017年
北海道大学、札幌

脂肪化肝細胞・脂肪肝に対するビルベリーの効果とその機序の検討　　　　　　　　　　　　　　　
芳賀 早苗, 荘厳 哲哉, 伊 敏, 森田 直樹, 浅野 真未, 尾崎 倫孝
第24回肝細胞研究会, 2017年
旭川市民文化会館、旭川

肝核内受容体FXRはp62/SQSTM1およびSHPを経由して、それぞれ抗酸化・細胞保護効果、脂肪化抑制効果を示す　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 伊 敏
第24回肝細胞研究会, 2017年
旭川市民文化会館、旭川

マウス脂肪化肝細胞・脂肪肝に対するビルベリーの抑制効果の検討　　　　　　　　　　　　　　　
芳賀 早苗, 荘厳 哲哉, 伊 敏, 森田 直樹, 浅野 真未, 尾崎 倫孝
第38回日本肥満学会, 2017年
大阪国際会議場、大阪

肝細胞におけるレドックス依存性ネクロプトーシスの動態解析　　　　　　　　　　　　　　　
芳賀 早苗, 菅野 憲, 小澤 岳昌, 森田 直樹, 浅野 真未, 伊 敏, 尾崎 倫孝
ConBio2017, 2017年
神戸国際会議場、神戸

ビルベリー抽出物の糖尿病性網膜症発症・進行予防効果に関する基礎的研究　　　　　　　　　　　　　　　
浅野 真未, 芳賀 早苗, 柴崎 彩, 黒澤 和也, 荘厳 哲哉, 尾崎 倫孝
ConBio2017, 2017年
神戸国際会議場、神戸

カスパーゼ3プローブ発現細胞を用いたクラミジア感染宿主細胞内のアポトーシス制御機構の探索　　　　　　　　　　　　　　　
松尾 淳司, 芳賀 早苗, 大久保 寅彦, 中村 眞二, 小澤 岳昌, 尾崎 倫孝, 山口 博之
ConBio2017, 2017年
神戸国際会議場、神戸

Optical evaluation and monitoring of oxidative stress & damage in vivo.　　　　　　　　　　　　　　　
Sanae Haga, Takeaki Ozawa, Naoki Morita, Mami Asano, James Remington, Michitaka Ozaki
19th International Symposium on Bioluminescence & Chemiluminescence(ISBC2016)., 2016年
茨城

Long-term ex vivo and in vivo monitoring of tumor progression by using dual luciferases.　　　　　　　　　　　　　　　
Naoki Morita, Sanae Haga, Yoshihiro Ohmiya, Michitaka Ozaki
19th International Symposium on Bioluminescence & Chemiluminescence (ISBC2016)., 2016年
茨城

Function of Toll-like receptor 2 in the murine inflammatory response to Rhodococcus aurantiacus infection.　　　　　　　　　　　　　　　
Yimin, Masashi Kohanawa, Sanae Haga, Michitaka Ozaki
4th Annual Meeting of the International Cytokine and Interferon Society (ICIS) ．, 2016年
米国

ビルベリーによる非アルコール性脂肪肝炎(NASH)の予防に向けた基礎的検討」（ポスター）　　　　　　　　　　　　　　　
芳賀 早苗, 荘厳 哲哉, 森田 直樹, 浅野 真未, 尾崎 倫孝
第23回肝細胞研究会, 2016年
大阪大学中之島センター、大阪

ビルベリーの糖尿病性網膜症初期病変抑制効果の検討　　　　　　　　　　　　　　　
芳賀早苗, 荘厳哲哉, 森田直樹, 浅野真未, 尾崎倫孝
第89回日本生化学会大会, 2016年
仙台国際センター、仙台

光プローブをもちいた時空間的病態解析と積極的病態制御の試み　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 小澤 岳昌, 山田 勇磨
第71回日本消化器外科学会総会 特別企画「光エネルギーの医療への展開」（, 2016年
あわぎんホール、徳島

FXRを起点とした脂肪肝傷害抑制と 脂肪化改善の機序　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 森田 直樹
第25回日本肝臓医生物学研究会；LBSG-J（プロメテウスの会）, 2016年
ナスパニューオータニ、新潟

「光技術を用いた臓器・細胞機能評価と制御の可能性」　　　　　　　　　　　　　　　
尾崎倫孝, 芳賀早苗, 小澤岳昌, 森田直樹, 浜田俊幸
第43回日本臓器保存生物医学会学術集会, 2016年
東京薬科大学、東京

“光”を利用した肝細胞機能制御技術の開発　　　　　　　　　　　　　　　
芳賀 早苗, 小澤 岳昌, 森田 直樹, 野田 なつみ, 尾崎 倫孝
第22回肝細胞研究会 ポスターセッション6「創薬・新しい実験系の開発」, 2015年
米子コンベンションセンター、鳥取

統合的ストレス応答による脂肪肝再生障害の分子メカニズムの解明　　　　　　　　　　　　　　　
稲葉 有香, 芳賀 早苗, 尾崎 倫孝, 春日 雅人, 井上 啓
第22回肝細胞研究会 一般口演5「肝再生」, 2015年
米子コンベンションセンター、鳥取

脂肪肝における肝切除後肝傷害・肝再生不全の機序について　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 小澤 岳昌, 森田 直樹, 井上 啓, 稲葉 有香
第22回肝細胞研究会, 2015年
米子コンベンションセンター、鳥取県

光技術を応用した移植臓器（組織）のモニタリングと制御の試み　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 森田 直樹, 伊 敏, 菅野 憲, 小澤 岳昌
第51回日本移植学会総会, 2015年
ホテル日航熊本、熊本県

Development of a novel in vivo optic imaging technology for biological diagnosis and therapy. (生物学的診断と治療のための新たな生体光イメージング技術の開発)　　　　　　　　　　　　　　　
尾崎倫孝, 芳賀早苗, 森田直樹, 伊 敏, 山田勇磨, 小澤岳昌
第74回 日本癌学会学術総会, 2015年
名古屋国際会議場、名古屋

p62/SQSTM1 を基軸とした新たな肝臓・肝細胞保護・機能維持法の探索　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 森田 直樹, 伊 敏
第42回日本臓器保存生物医学会学術集会、シンポジウム, 2015年
いわて県民情報交流センター、岩手

p62/SQSTM1が制御する肝細胞障害機序の解析　　　　　　　　　　　　　　　
芳賀 早苗, 小澤 岳昌, 山田 勇磨, 森田 直樹, 野田 なつみ, 尾崎 倫孝
第21回肝細胞研究会, 2014年
東京医科歯科大学、東京

高血糖および脂肪化状態が肝虚血/再灌流傷害に及ぼす影響 Impact of high glucose and steatosis upon ischemia/reperfusion-induced liver injury in mouse.　　　　　　　　　　　　　　　
芳賀 早苗, 小澤 岳昌, 森田 直樹, 野田 なつみ, 尾崎 倫孝
第87回日本生化学会, 2014年
京都国際会議場、京都

光イメージング技術を用いた肝病態の解析　　　　　　　　　　　　　　　
尾崎 倫孝, 芳賀 早苗, 野田 なつみ, 森田 直樹, 小澤 岳昌
第21回肝細胞研究会 シンポジウム1「肝疾患の病態を制御するメカニズム」, 2014年
東京医科歯科大学、東京

“ 光” を利用した新たな細胞・臓器保存法の開発　　　　　　　　　　　　　　　
芳賀 早苗, 小澤 岳昌, 森田 直樹, 野田 なつみ, 尾崎 倫孝
第41回日本臓器保存生物医学会学術集会 平成25 年度日本臓器保存生物医学会奨励賞 受賞記念講演, 2014年
千里ライフサイエンスセンター、大阪

異なる特性をもつ二種類の光を利用した生体内深部組織の修復・再生法の開発
科学研究費助成事業
2021年04月01日 - 2024年03月31日
尾崎 倫孝, 小澤 岳昌, 森田 直樹, 芳賀 早苗
日本学術振興会, 基盤研究(B), 北海道大学, 21H02983

薬理的神経制御を用いた新たな脳卒中運動療法の開発に対する生体脳イメージングの応用
科学研究費助成事業
2020年04月01日 - 2024年03月31日
前島 洋, 真先 敏弘, 榊間 春利, 高松 泰行, 芳賀 早苗, 尾崎 倫孝
昨年度までに得られた所見として、トレッドミル運動、或いはα5サブユニットを含むGABA受容体の特異的阻害薬L-655,708投与の単独介入による脳出血後の運動機能回復は限定的であるが、両者を組み合わせた介入により相乗的な回復効果が確認されていた。本年度は、これらの個体から採取した脳・脊髄サンプルの解析を進め、併用介入群に生じた可塑性修飾について検証を行った。L-655,708投与により大脳皮質運動野BDNF発現の増強が認められた。加えて、併用介入群の脊髄においては、 シナプスや軸索の可塑性に関与する分子マーカーの発現増強が認められた。このことから、併用介入群の効果的な機能回復には、脳内だけでなく脊髄における可塑性修飾も重要であることが示唆された。
脳におけるBDNF発現の生体イメージング計測を目的に、BDNFのプロモーター域に蛍発光酵素Luciferase遺伝子を挿入したBDNF-Luc Tgマウスを繁殖、飼育した。このTgマウスを対象に脳深部の発光の検知を可能とする発光基質AkaLumine-HCl (TokeOni)を腹腔内投与後、脳領域の発光を運動介入後に経時的に定量する一連の手法を開発した。この生体脳イメージングの手法を用いて、トレッドミル運動がBDNF発現に与える効果について検証を行った。単回のトレッドミル運動後４-8時間内において脳領域における発光、即ちBDNF発現が増強され、更に２週間のトレッドミル運動の継続することで運動後のより早いタイミングにBDNF発現増強が生じることを確認した。
一方、この生体脳イメージングを脳出血モデルに対して応用する際に、これまでのステレオタキシックシステムを用いた微量コラゲナーゼ注入によるモデル作成において、計測脳領域の皮膚処理が発光に影響を与える問題が生じ、術時における頭部の皮膚切開、縫合部位について検討を要することが示唆された。
日本学術振興会, 基盤研究(B), 北海道大学, 20H04048

細胞内Ca2+と活性酸素が誘導するプログラム細胞死による肝虚血再灌流傷害の新展開
科学研究費助成事業 基盤研究(C)
2020年04月 - 2024年03月
芳賀 早苗, 森田 直樹, 尾崎 倫孝
本研究は、虚血および再灌流時に肝細胞内で引き起こされる『活性酸素』, 『細胞内カルシウムイオン』, そして『（ネクローシス様）プログラム細胞死』に着目し、これまでの概念では説明しきれない肝傷害誘導機序の存在を明らかにすることを目的とする。この研究は、特に肝傷害の維持・拡大におおきく関わる、再灌流後中～後期の持続的な肝傷害をターゲットとし、新たな肝虚血再灌流傷害機構の解明に挑み、新たな標的分子・機序を導き出す意義を持つ。
本研究では、「ネクロプトーシス」、「パータナトス」など種々のネクローシス様プログラム細胞死等と、低酸素/再酸素化時に肝細胞内で引き起こされる様々なイベント、環境変化(①酸化ストレス、②細胞内カルシウムイオン(Ca2+)濃度―CaMKⅡ）の機序を段階的に進める計画である。
本年度は昨年度の研究成果を取りまとめるとともに、引き続き上記計画に則り研究を進めた。昨年度から着手している低酸素/再酸素化時の細胞内カルシウムイオン―CaMKⅡを介したネクロプトーシスの分子機序の解析を進め、この細胞死の機序や意義を解析した。これに関連して、ネクロプトーシスと他のプログラム細胞死との相互関係や相違についての解析も進めた。また、昨年度から取り掛かっている肝虚血/再灌流モデルにおける肝細胞死解析に関しては、ターゲットを抑制した条件での検討段階まで解析を進めた。また、各プログラム細胞死の重要性・機能性を検討するためのツールの開発に関して、事調査及び構想を固めることができた。
以上のように、本年度も肝細胞および生体肝で引き起こされる細胞死の機能性とその誘導機序について、基盤から応用までの検討を進めることができた。
日本学術振興会, 基盤研究(C), 北海道大学, 研究代表者, 20K08299

ランタニド・ナノ粒子（LNP）を利用した癌細胞特異的光治療法の開発　　　　　　　　　　　　　　　
挑戦的研究（開拓）
2019年04月 - 2023年03月
尾崎 倫孝
文部科学省, 競争的資金

肝虚血・再灌流傷害における多段階多元的傷害進展のメカニズム解析　　　　　　　　　　　　　　　
基盤研究(B)
2019年04月 - 2023年03月
森田 直樹
文部科学省, 競争的資金

生体細胞内分子の時空間的ダイナミズム解析のためのイメージング技術開発　　　　　　　　　　　　　　　
「医学系研究奨励（がん領域・基礎）」
2016年 - 2019年
芳賀早苗
公益財団法人 武田科学振興財団, 研究代表者, 競争的資金

分子標的治療薬の非侵襲的・時空間的モニタリングに向けた革新的イメージング技術開発　　　　　　　　　　　　　　　
科学研究費 若手研究（Ａ）
2015年 - 2019年
芳賀早苗
文部科学省, 研究代表者, 競争的資金

分子イメージングを基軸とする生細胞内分子計測・光操作法の開発　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ｓ）
2014年 - 2019年
小澤岳昌
文部科学省, 競争的資金

新規マーカーによるNASH予防・診断・治療のための食品・薬剤探索システムの構築　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ｂ）
2015年 - 2018年
森田直樹
文部科学省, 競争的資金

膵癌の機能的診断を目標とした新規バイオマーカーの開発　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2015年 - 2018年
増井俊彦
文部科学省, 競争的資金

膵癌における新たな細胞内分子ターゲットによる生物学的診断・治療法の開発　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2015年 - 2018年
森田直樹
文部科学省, 競争的資金

光による細胞機能制御による新規細胞療法の開発　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2014年 - 2017年
芳賀早苗
文部科学省, 研究代表者, 競争的資金

原始クラミジアが共生するアメーバは何故レジオネラの感染から回避できるのか　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2014年 - 2016年
山口博之
文部科学省, 競争的資金

細胞死による積極的な肝機能維持・再生制御機構の解明と臨床応用に向けた研究　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2014年 - 2016年
伊 敏
文部科学省, 競争的資金

多重ストレス時における精神活動変化の可視化とストレスマネジメント方略　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2013年 - 2016年
溝部佳代
文部科学省, 競争的資金

定量的生体分子イメージング・操作法の開発　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ａ）
2014年 - 2015年
小澤岳昌
文部科学省, 競争的資金

NAFLDに伴う肝再生障害に対する抗GADD34作用の有用性の検討　　　　　　　　　　　　　　　
科学研究費 若手研究（Ｂ）
2013年 - 2015年
稲葉有香
文部科学省, 競争的資金

肝ストレスの動的解析による肝機能障害・予備能の評価　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2012年 - 2014年
佐藤直樹
文部科学省, 競争的資金

生きた細胞内でのオートファジー発光定量法の開発　　　　　　　　　　　　　　　
科学研究費 若手研究（Ｂ）
2012年 - 2014年
服部満
文部科学省, 競争的資金

光プローブを応用した生体イメージング法による画期的術中ライブ診断法の開発　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ａ）
2011年 - 2014年
尾崎倫孝
文部科学省, 競争的資金

ヒスチジン誘導体の糖代謝における役割　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ｂ）
2011年 - 2014年
井上啓
文部科学省, 競争的資金

腹腔内転移のバイオイメージングと抑制物質の分子デザイン　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ｂ）
2011年 - 2014年
梅澤一夫
文部科学省, 競争的資金

細胞情報伝達に関わる蛋白質活性を可視化する発光プローブ分子の開発　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ｂ）
2011年 - 2014年
森田直樹
文部科学省, 競争的資金

心停止下肝移植への臨床応用をめざした肝グラフト灌流保存法の開発　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2011年 - 2012年
古川博之
文部科学省, 競争的資金

新規分泌型発光デュアルプローブを用いたｉｎ ｖｉｖｏ での癌細胞上皮間葉移行解析　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2011年 - 2012年
片岡昭彦
文部科学省, 競争的資金

細胞死(オートファジー・アポトーシス)による肝再生制御メカニズムの解析　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2011年 - 2012年
尾崎倫孝
文部科学省, 競争的資金

生体分子機能イメージングによる種々の肝病態におけるストレス応答の解析　　　　　　　　　　　　　　　
科学研究費 特別研究員奨励費
2010年 - 2012年
芳賀早苗
文部科学省, 研究代表者, 競争的資金

臓器ストレス測定法の開発と外科領域への応用　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2010年 - 2011年
藤堂省
文部科学省, 競争的資金

分泌型光プローブによる生体内リポーターシステムの開発と腫瘍診断・治療法への応用　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2009年 - 2010年
尾崎倫孝
文部科学省, 競争的資金

光プローブをもちいたバイオイメージングによる多角的診断・治療法の開発　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ａ）
2008年 - 2010年
尾崎倫孝
文部科学省, 競争的資金

分子機能プローブ導入による生体内臓器機能イメージングに関する研究　　　　　　　　　　　　　　　
科学研究費 特別研究員奨励費
2008年 - 2009年
芳賀早苗
文部科学省, 研究代表者, 競争的資金

生体材料(コラーゲンビトリゲル)をもちいた新しい治療材料の開発　　　　　　　　　　　　　　　
科学研究費 挑戦的萌芽研究
2008年 - 2009年
古川博之
文部科学省, 競争的資金

CHIP-chip法/Oligofish法によるヒトがん組織の網羅的転写機能解析　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ｃ）
2007年 - 2008年
中西一彰
文部科学省, 競争的資金

新規分子機能プローブ開発による生体・臓器内分子機能診断への応用　　　　　　　　　　　　　　　
科学研究費 萌芽研究
2007年 - 2008年
尾崎倫孝
文部科学省, 競争的資金

生活関連化学物質の皮膚感作性等のインビトロ評価法に関する研究　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ｃ）
2006年 - 2008年
内野正国
文部科学省, 競争的資金

消化器外科領域における分子標的治療に向けた包括的基盤研究　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ｂ）
2005年 - 2007年
尾崎倫孝
文部科学省, 競争的資金

肝移植代替治療をめざした胎児肝内への同種・異種細胞移植の生着・増殖条件の検討　　　　　　　　　　　　　　　
科学研究費 基盤研究（Ｃ）
2001年 - 2002年
絵野沢伸
文部科学省, 競争的資金