分子シャペロンによる細胞内ストレス制御の分子基盤解明とその糖尿病治療への応用

2013 Fiscal Year Annual Research Report

• Project/Area Number: 23390243
• Research Institution: Kumamoto University
• Principal Investigator: 荒木 栄一 熊本大学, 生命科学研究部, 教授 (10253733)
• Co-Investigator(Kenkyū-buntansha): 近藤 龍也 熊本大学, 生命科学研究部, 助教 (70398204)
• Project Period (FY): 2011-11-18 – 2014-03-31
• Keywords: 糖尿病 / 小胞体ストレス

Research Abstract

研究１（in vitro）インスリンの標的臓器である肝臓(HepG2)、筋肉(C2C12)、脂肪(3T3-L1)、およびインスリン産生細胞である膵β細胞(MIN6)の各々の細胞株において分子シャペロンであるBiPもしくはHSP72を過剰発現あるいはsiRNAを用いてノックダウンした。BiPを導入した細胞群においては、明らかなインスリンシグナルの改善は認めなかったが、HSP72を過剰発現させた細胞では高血糖環境下でのインスリン作用低下現象が回復した。また小胞体ストレス耐性に関してはBiP, HSP72どちらの導入も、tunicamycinやthapsigargin刺激における抵抗性を示した。逆にノックダウン系では、反対の現象、すなわちインスリン感受性の悪化と小胞体ストレス耐性の低下を認めた。遺伝子発現解析にては、AMPK, PGC-1a, Sirt1の発現増加を認めた。
研究２（ in vivo)　RIP (rat insulin promotor)あるいはalbuminの下流で変異リガンド結合ドメインを持つエストロゲン受容体下にCreを発現できるトランスジェニックマウスを作成し、系統を維持に至った。さらにCreの作用でCAT配列をを削除し、BiPもしくはHSP72を時期・組織特異的に過剰発現できるfloxマウスも作成した。これらのマウスの交配を行い、糖尿病を誘発する刺激（高脂肪食・高フルクトース食・GTGによる摂食中枢破壊肥満モデルなど）を行って、耐糖能・インスリン抵抗性などの代謝レベルの実験を行っていく予定である。

Current Status of Research Progress

Reason

25年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

25年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] The serine protease prostasin regulates hepatic insulin sensitivity by modulating TLR4 signalling. (2014)
  • Author(s): Uchimura K, Hayata M, Mizumoto T, Miyasato Y, Kakizoe Y, Morinaga J, Onoue T, Yamazoe R, Ueda M, Adachi M, Miyoshi T, Shiraishi N, Ogawa W, Fukuda K, Kondo T, Matsumura T, Araki E, Tomita K, Kitamura K.
  • Journal Title: Nature Communications Volume: 5 Pages: 3428
  • DOI: 10.1038/ncomms4428.
  • Peer Reviewed
• [Journal Article] Prevalence of metabolic syndrome in stroke patients: a prospective multicenter study in Japan. (2014)
  • Author(s): Takahashi T, Harada M, Kikuno T, Ujihara M, Sadamitsu D, Manabe Y, Yasaka M, Takayama H, Kobori S, Araki E
  • Journal Title: Acute Med Surg Volume: 1 Pages: 17-22
  • Peer Reviewed
• [Journal Article] for the Tofogliflozin 004 and 005 Study group. Long-term safety and efficacy of tofogliflozin, a selective inhibitor of sodium-glucose cotransporter 2, as monotherapy or in combination with other oral antidiabetic agents in Japanese patients with type 2 diabetes mellitus: multicenter, open-label, randomized controlled trials (2014)
  • Author(s): Tanizawa Y, Kaku K, Araki E, Tobe K, Terauchi Y, Utsunomiya K, Iwamoto Y, Watada H, Ohtsuka W, Watanabe D, Suganami H
  • Journal Title: Expert Opin Pharmacother Volume: 15(6) Pages: 749-766
  • DOI: 10.1517/14656566.2014.887680.
  • Peer Reviewed
• [Journal Article] Association between circulating leukocyte subtype counts and carotid intima-media thickness in Japanese subjects with type 2 diabetes. (2013)
  • Author(s): Matsumura T, Taketa K, Motoshima H, Senokuchi T, Ishii N, Kinoshita H, Fukuda K, Yamada S, Kukidome D, Kondo T, Hisada A, Katoh T, Shimoda S, Nishikawa T, Araki E.
  • Journal Title: Cardiovascular Diabetology Volume: 12(1) Pages: 177
  • DOI: 10.1186/1475-2840-12-177.
  • Peer Reviewed
• [Journal Article] Effects of combination therapy with vildagliptin and valsartan in a mouse model of type 2 diabetes. (2013)
  • Author(s): Miyagawa K, Kondo T, Goto R, Matsuyama R, Ono K, Kitano S, Kawasaki S, Igata M, Kawashima J, Matsumura T, Motoshima H, Araki E.
  • Journal Title: Cardiovascular Diabetology Volume: 12(1) Pages: 160
  • DOI: 10.1186/1475-2840-12-160.
  • Peer Reviewed
• [Journal Article] Efficacy and safety of sitagliptin as add-on therapy on glycemic control and blood glucose fluctuation in Japanese type 2 diabetes subjects ongoing with multiple daily insulin injections therapy. (2013)
  • Author(s): Shimoda S, Iwashita S, Ichimori S, Matsuo Y, Goto R, Maeda T, Matsuo T, Sekigami T, Kawashima J, Kondo T, Matsumura T, Motoshima H, Furukawa N, Nishida K, Araki E.
  • Journal Title: Endocrine Journal Volume: 60(10) Pages: 1207-1214
  • DOI: 10.1507/endocrj.EJ13-0198
  • Peer Reviewed
• [Journal Article] Insulin receptor activation through its accumulation in lipid rafts by mild electrical stress. (2013)
  • Author(s): Morino-Koga S, Yano S, Kondo T, Shimauchi Y, Matsuyama S, Okamoto Y, Suico MA, Koga T, Sato T, Shuto T, Arima H, Wada I, Araki E, Kai H
  • Journal Title: J Cell Physiol Volume: 228(2) Pages: 439-446
  • DOI: 10.1002/jcp.24149.
  • Peer Reviewed
• [Journal Article] Evaluation of a new device for measurement of hemoglobin A1c for Japanese subjects. (2013)
  • Author(s): Shimoda S, Maeda T, Furukawa N, Ichinose K, Taketa K, Igata M, Senokuchi T, Matsumura T, Araki E
  • Journal Title: Diabetology International Volume: 4 Pages: 112-116
  • Peer Reviewed
• [Journal Article] Relationship between lifestyle and lifestyle-related factors in a rural-urban population of Japan. (2013)
  • Author(s): Zhang SC, Wei CN, Harada K, Ueda K, Fukumoto K, Matsuo H, Minamoto K, Nishikawa T, Araki E, Ueda A, Fang J
  • Journal Title: Environ Health Prev Med Volume: 18 Pages: 267-274
  • DOI: 10.1007/s12199-012-0315-5.
  • Peer Reviewed
• [Journal Article] Long-term safety of linagliptin monotherapy in Japanese patients with type 2 diabetes. (2013)
  • Author(s): Araki E, Kawamori R, Inagaki N, Watada H, Hayashi N, Horie Y, Sarashina A, Thiemann S, von Eynatten M, Dugi K, Woerle HJ
  • Journal Title: Diabetes Obes Metab Volume: 15 Pages: 364-371
  • DOI: 10.1111/dom.12039
  • Peer Reviewed
• [Journal Article] The phosphatase Dullard negatively regulates BMP signalling and is essential for nephron maintenance after birth. (2013)
  • Author(s): Sakaguchi M, Sharmin S, Taguchi A, Ohmori T, Fujimura S, Abe T, Kiyonari H, Komatsu Y, Mishina Y, Asashima M, Araki E, Nishinakamura R
  • Journal Title: Nature Communications Volume: 4 Pages: 1398
  • DOI: 10.1038/ncomms2408.
  • Peer Reviewed
• [Journal Article] Mechanism-based antioxidant therapies promise to prevent diabetic complications? (2013)
  • Author(s): Nishikawa T, Araki E
  • Journal Title: Diabetes Invest Volume: 4 Pages: 105-107
  • Peer Reviewed
• [Presentation] Heat Treatment with Mild Electrical Stimulation Attenuates Systemic Inflammation and Reduces Cytokine Expression in Monocytes from subjects with Metabolic Syndrome (2014)
  • Author(s): Tatsuya Kondo, Rina Matsuyama, Rieko Goto, Kaoru Ono, Sayaka Kitano, Junji Kawashima, Hiroyuki Motoshima, Takeshi Matsumura, Hirofumi Kai, Eiichi Araki
  • Organizer: The 74th Annual Meeting of American Diabetes Association
  • Place of Presentation: San Francisco, USA
  • Year and Date: 20140613-20140617
• [Presentation] Activation of Heat Shock Response Improves Metabolic Profiles in Type 2 Diabetes. (2013)
  • Author(s): Tatsuya Kondo, Katsutoshi Miyagawa, Rina Matsuyama, Rieko Goto, Kaoru Ono, Sayaka Kitano, Hiroyuki Motoshima, Junji Kawashima, Takeshi Matsumura, Hirofumi Kai, Eiichi Araki.
  • Organizer: 2013 International Conference of Diabetes and Metabolism and 5th AASD
  • Place of Presentation: Seoul, Korea
  • Year and Date: 20131106-20131109
  • Invited
• [Presentation] 熱ショック応答経路活性化による糖代謝改善効果と分子機構の解明 (2013)
  • Author(s): 近藤龍也, 荒木栄一
  • Organizer: 第63回日本体質医学会
  • Place of Presentation: 久留米、久留米大学筑水会館
  • Year and Date: 20131005-20131006
• [Presentation] Pain threshold values of small fibers are associated with the severity of diabetic complications. (2013)
  • Author(s): Kukidome D, Sato M, Shimoda S, Nishikawa T, Araki E
  • Organizer: 49th EASD Annual Meeting
  • Place of Presentation: Barcelona,Spain
  • Year and Date: 20130923-20130927
• [Presentation] Heat Shock Response Activation using Heat Treatment with Mild Electrical Stimulation Reduces Visceral Adiposity and Improves Insulin Resistance and Inflammation in Male Subjects with Type 2 Diabetes (2013)
  • Author(s): Tatsuya Kondo, Rina Matsuyama, Rieko Goto, Kaoru Ono, Sayaka Kitano, Junji Kawashima, Hiroyuki Motoshima, Takeshi Matsumura, Hirofumi Kai, Eiichi Araki
  • Organizer: CSSI scientific meeting.
  • Place of Presentation: Sheffield, United Kingdom
  • Year and Date: 20130818-20130822
• [Presentation] The Role of Aldosterone in the Pathogenesis of Diabetes in Human and Rats. (2013)
  • Author(s): Goto R, Kondo T, Ono K, Matsuyama R, Miyagawa K, Igata M, Kawashima J, Matsumura T, Motoshima H, Shimoda S, Araki E
  • Organizer: The 73th Annual Meeting of American Diabetes Association
  • Place of Presentation: Chicago, USA
  • Year and Date: 20130621-20130625
• [Presentation] The Critical Role of Heat Shock Protein 72 in Diabetic Pathophysiology. (2013)
  • Author(s): Matsuyama R, Kondo T, Miyagawa K, Goto R, Ono K, Igata M, Kawashima J, Matsumura T, Motoshima H, Araki E
  • Organizer: The 73th Annual Meeting of American Diabetes Association
  • Place of Presentation: Chicago, USA
  • Year and Date: 20130621-20130625
• [Presentation] 酸化ストレスからみた合併症治療の展望 (2013)
  • Author(s): 西川武志, 佐田公範, 久木留大介, 荒木栄一
  • Organizer: 第56回日本糖尿病学会年次学術集会
  • Place of Presentation: 熊本、ホテル日航熊本
  • Year and Date: 20130516-20130518
• [Presentation] Activation of Heat Shock Response ameliorates Metabolic Abnormalities in Diabetes with Attenuation of Cell Stress (2013)
  • Author(s): 近藤龍也, 荒木栄一
  • Organizer: 第56回日本糖尿病学会年次学術集会
  • Place of Presentation: 熊本、ホテル日航熊本
  • Year and Date: 20130516-20130518
• [Book] 2型糖尿病. 糖尿病最新の治療2013-2015(岩本安彦, 羽田勝計編) (2013)
  • Author(s): 荒木栄一
  • Total Pages: 366
  • Publisher: 南江堂
• [Book] 特集：肥満症 2型糖尿病と耐糖能異常 (2013)
  • Author(s): 古川昇, 荒木栄一
  • Total Pages: 169
  • Publisher: 日本臨床
• [Book] PPARγの最新動向 PPAR標的薬の開発動向. 期待されるチアゾリジン薬 (2013)
  • Author(s): 松村剛, 荒木栄一
  • Total Pages: 308
  • Publisher: フジメディカル出版
• [Remarks] 熊本大学大学院生命科学研究部代謝内科学
  • URL: http://www2.kuh.kumamoto-u.ac.jp/met/