病理学の最新テクノロジーを応用した漢方薬の薬効・機能解析法の確立

2014 Fiscal Year Annual Research Report

• Project/Area Number: 24390181
• Research Institution: The University of Tokushima
• Principal Investigator: 常山 幸一 徳島大学, ヘルスバイオサイエンス研究部, 教授 (10293341)
• Co-Investigator(Kenkyū-buntansha): 藤本 誠 富山大学, 大学病院, 講師 (30377337) ; 平 修 福井県立大学, 生物資源学部, 准教授 (30416672)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: メタボリックシンドローム / NASH / 動物モデル / 漢方薬 / 質量顕微鏡

Outline of Annual Research Achievements

メタボリックシンドローム(MetS)の肝での表現型であるNAFLD/NASH は21 世紀の肝臓病とも呼ばれ、先進諸国で患者数が急増してい る。世界中で有効な予防法や治療法が研究されているが、いまだ有効な薬物は確立されていない。我々は、漢方薬や生薬は食に近く安 全に長期摂取が可能であり、MetS やNASH の予防的投与に最適な薬物と考えている。本研究では、独自に開発したMetS・NASH モデル マウスと生薬に含まれる種々の低分子化合物の 局在を細胞レベルで可視化できる新技術(質量顕微鏡)を応用し、動物モデルの各臓器における化合物・代謝産物の局在を明らかにするとともに、それら化合物が直接コンタクトしている標的細胞を同定し、薬剤の作用機序の解明を試みる。本年度は、従来のモデルよりも、よりヒトの病態に近いMetS/NASHモデル動物を開発した。このモデルを用いることで、これまでに検討ができなかった、肝線維化のメカニズムや肝発癌のメカニズム、さらに、糖尿病発症と肝発癌との相関のより詳細、かつ経時的な解析が可能となった。さらに、これら動物モデルの病理組織学データを同様の病態のヒト組織像と相関させ、スコア化によって半定量的に比較検討できる実験系を確立した。また、鉄をコアにしたナノ粒子のアシストによって低分子物質をイオン化し、マウス肝組織中で質量顕微鏡にてイメージング解析することに成功した。これらの新規動物モデルと質量分析による新たなイメージング法を組み合わせ、防已黄耆湯などの和漢薬やクロロゲン酸などのポリフェノール、コレステロールや各種胆汁酸成分等の作用機序の解明を試みるとともに、標的物質の局在や、周囲の免疫学的な微小環境を明らかにした。これらの解析結果は国内外の学会で報告するとともに、欧文誌に発表を行った（一部は現在論文作成中、あるいは投稿中）。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Causes of Carryover

26年度が最終年度であるため、記入しない。

Expenditure Plan for Carryover Budget

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Carbamazepine-Induced Liver Injury Requires CYP3A-Mediated Metabolism and Glutathione Depletion in Rats. (2015)
  • Author(s): Iida A, Sasaki E, Yano A, Tsuneyama K, Fukami T, Nakajima M, Yokoi T.
  • Journal Title: Drug Metab Dispos. Volume: in press Pages: in press
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] An SHR/NDmcr-cp Rat Model of Non-alcoholic steatohepatitis with advanced fibrosis induced by a High-fat, High-cholesterol Diet. (2015)
  • Author(s): Ichimura M, Hatanaka M, Tsuneyama K, Kato S, Omagari K.
  • Journal Title: J Obes Weight Loss There Volume: in press Pages: in press
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Emerging of IL-33 in inflammation and immune regulation. (2015)
  • Author(s): Ikutani M, tsuneyama K, Nakae S, Takatsu K.
  • Journal Title: Inflammation and Regeneration Volume: 35 Pages: 69-77
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Effect of keishibukuryogan on genetic and dietary obesity models. (2015)
  • Author(s): Gao F, Yokoyama S, Fujimoto M, Tsuneyama K, Saiki I, Shimada Y, Hayakawa Y.
  • Journal Title: Evid Based Complement Alternat Med. Volume: in press Pages: in press
  • DOI: 10.1155/2015/801291.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Immunologic potential of CTLA-4 Ig in murine autoimmune cholangitis. (2015)
  • Author(s): Tanaka H, Yang GX, Tomiyama T, Tsuneyama K, Zhang W, Leung PS, Coppel RL, Joh T, Nadler SG, Ansari AA, Bowlus C, Gershwin ME.
  • Journal Title: Clin Exp Immunol. Volume: in press Pages: in press
  • DOI: 10.1111/cei.12581
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] igh-fat and high-cholesterol diet rapidly induces non-alcoholic steatohepatitis with advanced fibrosis in Sprague-Dawley rats. (2015)
  • Author(s): Ichimura M, Kawase M, Masuzumi M, Sakaki M, Nagata Y, Tanaka K, Suruga K, Tamaru S, Kato S, Tsuneyama K, Omagari K.
  • Journal Title: Hepatol Res. Volume: 45 Pages: 458-469
  • DOI: 10.1111/hepr.12358.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] CXCL16 suppresses liver metastasis of colorectal cancer by promoting TNF-α-induced apoptosis by tumor-associated macrophages. (2014)
  • Author(s): Kee JY, Ito A, Hojo S, Hashimoto I, Igarashi Y, Tsuneyama K, Tsukada K, Irimura T, Shibahara N, Takasaki I, Inujima A, Nakayama T, Yoshie O, Sakurai H, Saiki I, Koizumi K.
  • Journal Title: BMC Cancer. Volume: 14 Pages: 949
  • DOI: 10.1186/1471-2407-14-949.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Role of cytochrome P450-mediated metabolism and identification of novel thiol-conjugated metabolites in mice with phenytoin-induced liver injury. (2014)
  • Author(s): Sasaki E, Iwamura A, Tsuneyama K, Fukami T, Nakajima M, Kume T, Yokoi T.
  • Journal Title: Toxicol Lett. Volume: 232 Pages: 79-88
  • DOI: 10.1016/j.toxlet.2014.10.012
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Clinicopathological analysis of CD8-positive lymphocytes in the tumor parenchyma and stroma of hepatocellular carcinoma. (2014)
  • Author(s): An JL, Ji QH, An JJ, Masuda S, Tsuneyama K.
  • Journal Title: Oncol Lett. Volume: 8 Pages: 2284-2290
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Isoliquiritigenin is a potent inhibitor of NLRP3 inflammasome activation and diet-induced adipose tissue inflammation. (2014)
  • Author(s): Honda H, Nagai Y, Matsunaga T, Okamoto N, Watanabe Y, Tsuneyama K, Hayashi H, Fujii I, Ikutani M, Hirai Y, Muraguchi A, Takatsu K.
  • Journal Title: J Leukoc Biol. Volume: 96 Pages: 1087-1100
  • DOI: 10.1189/jlb.3A0114-005RR
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Differential modulation by IL-17A of Cholangitis versus Colitis in IL-2Rα deleted mice. (2014)
  • Author(s): Yang W, Yao Y, Yang YQ, Lu FT, Li L, Wang YH, Nakajima T, Tsuneyama K, Ridgway WM, Gershwin ME, Lian ZX.
  • Journal Title: PLoS One. Volume: 9 Pages: e105351
  • DOI: 10.1371/journal.pone.0105351
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Animal models of primary biliary cirrhosis. (2014)
  • Author(s): Wang J, Yang GX, Tsuneyama K, Gershwin ME, Ridgway WM, Leung PS. Animal models of primary biliary cirrhosis.
  • Journal Title: Semin Liver Dis. Volume: 34 Pages: 285-296
  • DOI: 10.1055/s-0034-1383728
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Successful immunotherapy of autoimmune cholangitis by adoptive transfer of forkhead box protein 3(+) regulatory T cells. (2014)
  • Author(s): anaka H, Zhang W, Yang GX, Ando Y, Tomiyama T, Tsuneyama K, Leung P, Coppel RL, Ansari AA, Lian ZX, Ridgway WM, Joh T, Gershwin ME.
  • Journal Title: Clin Exp Immunol. Volume: 178 Pages: 253-261
  • DOI: 10.1111/cei.12415
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Changes in the expression of miRNAs at the pericentral and periportal regions of the rat liver in response to hepatocellular injury: comparison with the changes in the expression of plasma miRNAs. (2014)
  • Author(s): Yamaura Y, Nakajima M, Tatsumi N, Takagi S, Fukami T, Tsuneyama K, Yokoi T.
  • Journal Title: Toxicology. Volume: 322 Pages: 89-98
  • DOI: 10.1016/j.tox.2014.05.008.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Retinoid X receptor α in human liver is regulated by miR-34a. (2014)
  • Author(s): Oda Y, Nakajima M, Tsuneyama K, Takamiya M, Aoki Y, Fukami T, Yokoi T.
  • Journal Title: Biochem Pharmacol. Volume: 90 Pages: 179-187
  • DOI: 10.1016/j.bcp.2014.05.002.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Induction of insulin secretion by apolipoprotein M, a carrier for sphingosine 1-phosphate. (2014)
  • Author(s): Kurano M, Hara M, Tsuneyama K, Sakoda H, Shimizu T, Tsukamoto K, Ikeda H, Yatomi Y.
  • Journal Title: Biochim Biophys Acta. Volume: 1841 Pages: 1217-1226
  • DOI: 10.1016/j.bbalip.2014.05.002.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Malignant transformation of hyperplastic gastric polyps: An immunohistochemical and pathological study of the changes of neoplastic phenotype. (2014)
  • Author(s): Imura J, Hayashi S, Ichikawa K, Miwa S, Nakajima T, Nomoto K, Tsuneyama K, Nogami T, Saitoh H, Fujimori T.
  • Journal Title: Oncol Lett. Volume: 7 Pages: 1459-1463
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Neonatal monosodium glutamate treatment causes obesity, diabetes, and macrovesicular steatohepatitis with liver nodules in DIAR mice. (2014)
  • Author(s): Tsuneyama K, Nishida T, Baba H, Taira S, Fujimoto M, Nomoto K, Hayashi S, Miwa S, Nakajima T, Sutoh M, Oda E, Hokao R, Imura J.
  • Journal Title: J Gastroenterol Hepatol. Volume: 29 Pages: 1736-1743
  • DOI: 10.1111/jgh.12610
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Neonatal streptozotocin treatment causes type 1 diabetes and subsequent hepatocellular carcinoma in DIAR mice fed a normal diet. (2014)
  • Author(s): Baba H, Tsuneyama K, Nishida T, Hatta H, Nakajima T, Nomoto K, Hayashi S, Miwa S, Nakanishi Y, Hokao R, Imura J.
  • Journal Title: Hepatology International Volume: 8 Pages: 415-424
  • DOI: 10.1007/s12072-014-9541-9
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Molecular mechanisms of hyperthermia-induced apoptosis enhanced by docosahexaenoic acid: implication for cancer therapy. (2014)
  • Author(s): Cui ZG, Piao JL, Kondo T, Ogawa R, Tsuneyama K, Zhao QL, Feril LB Jr, Inadera H.
  • Journal Title: Chem Biol Interact. Volume: 215 Pages: 46-53
  • DOI: 0.1016/j.cbi.2014.03.005.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Involvement of immune- and inflammatory-related factors in flucloxacillin-induced liver injury in mice. (2014)
  • Author(s): Takai S, Higuchi S, Yano A, Tsuneyama K, Fukami T, Nakajima M, Yokoi T.
  • Journal Title: J Appl Toxicol. Volume: 35 Pages: 142-151
  • DOI: 10.1002/jat.3002.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Distinct from its canonical effects, deletion of IL-12p40 induces cholangitis and fibrosis in interleukin-2Rα(-/-) mice. (2014)
  • Author(s): Yao Y, Yang W, Yang YQ, Ma HD, Lu FT, Li L, Tao YY, Tsuneyama K, Zhang W, Friedman S, Gershwin ME, Lian ZX.
  • Journal Title: J Autoimmun. Volume: 51 Pages: 99-108
  • DOI: 10.1016/j.jaut.2014.02.009.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Social isolation stress-induced fear memory deficit is mediated by down-regulated neuro-signaling system and Egr-1 expression in the brain. (2014)
  • Author(s): Okada R, Matsumoto K, Tsushima R, Fujiwara H, Tsuneyama K.
  • Journal Title: Neurochem Res. Volume: 39 Pages: 875-882
  • DOI: 10.1007/s11064-014-1283-5.
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] Daily moderate coffee intake inhibits pancreatic beta-cell damage and nonalcoholic steatohepatitis without improving obesity in a mouse model of spontaneous metabolic syndrome (2015)
  • Author(s): Koichi Tsuneyama, Hayato Baba, Takeshi Nishida, Hideki Hatta, Johji Imura
  • Organizer: The 24th Conference of the APASL
  • Place of Presentation: Istanbul (Turkey)
  • Year and Date: 2015-03-12 – 2015-03-15
• [Presentation] TSODマウスの有する新たな可能性を探る (2015)
  • Author(s): 常山幸一、渡邊俊介、高橋徹行、小川博久、上原久典、西田健志、八田秀樹、馬場逸人、井村穣二
  • Organizer: 第１０回TSODマウス研究会
  • Place of Presentation: つくば国際会議場（茨城県つくば市）
  • Year and Date: 2015-03-06
  • Invited
• [Presentation] 見えないものを“見る”技術～メタボリックシンドロームからイタイイタイ病まで～ (2015)
  • Author(s): 常山幸一
  • Organizer: 徳島臨床細胞学会
  • Place of Presentation: とくしま未来健康づくり機構（徳島県徳島市）
  • Year and Date: 2015-03-05
• [Presentation] Establishment of repeated liver biopsy technique in experimental mice (2014)
  • Author(s): Tsuneyama K, Baba H, Nakajima T, Nishida T, Imura J.
  • Organizer: 65th Annual meeting of the AASLDT,2014/11/7-11
  • Place of Presentation: Boston (USA)
  • Year and Date: 2014-11-07 – 2014-11-11
• [Presentation] Nano-PALDIイメージング質量分析を用いた低分子Xenobiotics（２オクチン酸）検出法の開発 (2014)
  • Author(s): 常山幸一、菊池健太郎、守時由起、井村穣二、平修
  • Organizer: JDDW 2014
  • Place of Presentation: 神戸国際展示場（兵庫県神戸市）
  • Year and Date: 2014-10-23 – 2014-10-26
• [Presentation] イメージングMSの病理診断学への応用と展開 (2014)
  • Author(s): 常山 幸一、井村 穣二
  • Organizer: 第４１回BMSコンファレンス
  • Place of Presentation: 能登ロイヤルホテル（石川県羽咋郡）
  • Year and Date: 2014-07-07 – 2014-07-09
• [Presentation] DIAR-MSGマウス：標準食飼育で肥満・糖尿病を経て大滴性脂肪性肝炎～肝細胞癌に至る新規モデル動物の開発 (2014)
  • Author(s): 常山幸一、西田健志、野本一博、林伸一、三輪重治、中島隆彦、中西ゆう子、八田秀樹、井村穣二
  • Organizer: 第１回肝臓と糖尿病代謝研究会
  • Place of Presentation: 伊藤国際学術研究センター（東京都）
  • Year and Date: 2014-07-04
• [Presentation] 自然発症メタボリックシンドローム-発癌モデル（TSODマウス）の肝腫瘍の特徴 (2014)
  • Author(s): 常山 幸一、西田 健志、野本一博、林伸一、三輪重治、中嶋隆彦、八田秀樹、井村穣二
  • Organizer: 第１０３回日本病理学会総会
  • Place of Presentation: 広島国際会議場（広島県広島市）
  • Year and Date: 2014-04-24 – 2014-04-26