1994 Fiscal Year Annual Research Report

糖尿病における高インスリン血症と動脈硬化-血管平滑筋細胞を用いた分子生物学的研究

Research Project

Project/Area Number	06454337
Research Institution	Shiga University of Medical Science
Principal Investigator	繁田幸男滋賀医科大学, 医学部, 教授 (60028321)
Co-Investigator(Kenkyū-buntansha)	日高秀樹滋賀医科大学, 医学部, 助手 (80156603) 柏木厚典滋賀医科大学, 医学部, 講師 (20127210)
Keywords	高インスリン血症 / 血管平滑筋細胞 / 糖尿病 / インスリン抵抗性 / 脱感作 / IRS-1 / インスリン作用 / 動脈硬化症
Research Abstract	多くの疫学的研究によると、高インスリン血症は冠動脈硬化症の危険因子として確立されている。しかしその血管平滑筋細胞増殖に関する直接的証拠は、インスリン結合と非生理的インスリン濃度でのDNA合成のみ検討されているだけである。我々はこれまでインスリン作用の分子生物学的研究及び糖尿病における高インスリン血症と高血糖の血管障害作用を血管細胞生物学的手法を用いて研究してきた(参考論文参照)。そこで本研究では、インスリンの作用機構に関して、インスリン受容体を過剰発現したrat-1線維芽細胞を用いてインスリン情報伝達機構を明らかにし、更に培養血管平滑筋細胞を用いて特異的インスリン受容体情報伝達系とその異常を検討し、高インスリン血症と動脈硬化症発症との関連を血管分子生物学的手法を用いて明らかにした。 (I)培養血管平滑筋細胞でのインスリン情報伝達系の活性化と細胞増殖:1)生理的インスリン濃度で、培養血管平滑筋細胞のインスリン受容体は自己燐酸化され、IRS-1のチロシン燐酸化を認めた。一方Shcのチロシン燐酸化を認めなかった。その結果生理的インスリン濃度ではMAPkinase、p90RSKの活性化は起こらず、DNA合成の誘導を認めなかった。2)一方生理的濃度のインスリンで、IRS-1に連結したPI3kinase、p70S6kinaseの活性化を認め、AIBの輸送は刺激された。このように平滑筋におけるインスリン作用はダンパンがIGF-1と同様に細胞増殖を促進するかどうかを更に明らかにする。 (II)高インスリン血症による血管平滑筋細胞におけるインスリン作用の脱感作:生理的濃度の高インスリン処理によって血管平滑筋細胞のインスリン結合は低下し、インスリンによる自己燐酸化、p70S6kinaseの活性化が低下した。すなわちインスリン抵抗性が誘導された。 (III)in vivo大動脈平滑筋細胞のインスリンシグナル伝達系と血管機能を検討している。 (IV)高インスリン血症インスリン抵抗性糖尿病モデル動物であるOLETFラットでの大動脈平滑筋細胞におけるインスリン情報伝達機構異常、各種遺伝子発現異常を検討し、高インスリン血症と動脈硬化の初期病変との関連を明らかにする。

Research Products

(20 results)

All Other

All Publications (20 results)

[Publications] Ugi ct al.: "Src homology 2 domains of protcin tyrosinc phosphatasc arc associatcd in vitro with both the insulin receptor and insulin receptor substratc-1 via different phosphotyrosinc motifs." FEBS Lcttcr. 340. 216-220 (1994)
[Publications] Maegawa et al.: "Insulin receptor kinase phosphorylates tyrosine phosphatasc containing Src homology 2 region and modulates its PTPase activity in vitro." Biochem Biophys Res Commun. 199. 780-785 (1994)
[Publications] Yasuda et al.: "Mctabolic effect of PGEI analoguc 01206.aCD on ncrvc Na+-K+-ATPasc activity of rais with streptozocin-induccd diabetes is mcdiated via cAMP:Possible role of cAMP in diabetic neuropathy." Prostaglandins. 47. 367-378 (1994)
[Publications] Haneda et al.: "Intcraction between atrial natriurctic peptide and angiotensin II cultured glomcrular mesangial cells and its disturbancc in diabetes." ExP.Nephrol. 2. 106-106 (1994)
[Publications] Kashiwagi et al.: "Abnormal glutathionc mctabolism and increascd cytotoxicity H202 in human umbilical vein endothclial cells culturcd in high glucose medium." Diabetologia. 37. 264-269 (1994)
[Publications] Ide et al.: "High glucose condition activates protein tyrosine phosphatase and deactivates insulin receptor function in insulinscnsitivc rat fibroblasts." Biochem Biophys Res Commun. 201. 71-77 (1994)
[Publications] Ide et al.: "High Glucose Concentration dcsensitizes insulin action at the levcls of receptor kinase." Endocrine J.42. 1-8 (1995)
[Publications] Kikkawa et al.: "Translocation of protein kinaseCα and in rat glomerular mesangial cells cultured under high glucose conditions." Diabetologia. 37. 838-841 (1994)
[Publications] Shigeta et al.: "Diabetic nephropathy in Japan." Diabetic Research and Clinical Practice. 24. S191-S197 (1994)
[Publications] Hidaka et al.: "Effects of lipo prostaglandin E1 injection on transcutancous oxygen tension in diabetic subjects with periphcral vascular discase." J Japanese Collage of Angiology. 34. 433-437 (1994)
[Publications] Takagi et al: "Insulin receptor-mediated signal transduction and its desensitization in culturcd vascular smooth musele cells." Atherosclerosis. 113. 19-27 (1995)
[Publications] Asahina et al.: "Impaired activation of glucose oxidation and NADPH supply in human endothelial cells exposed to H202 in hith glucose medium" Diabctes. (in press).
[Publications] Hidaka et al.: "Effects of an HMG-CoA reductase inhibitor,pravastatin,and bile sequestering resin,cholestyramine,on plasma plant sterol Levels in hypercholesterolemic subjects." Atherosclerosis Thromb. (in press).
[Publications] Sakamoto et al.: "Prevention of glomerular hyperliltration in rats with streptozotocin-induced diabetes by an atrial natriuretic peptide receptor antagonist." Diabetologia. (in press).
[Publications] Takigawa et al.: "Antibodies against GM,anglioside affect K+and N+currents in isolated rat myelinated nerve fibers" Ann Neurol. (in press).
[Publications] Takagi et al.: "Significance of fructose autoxidation protein oxidation and formation of advanced glycation end product." J Diabetes Compl. (in press).
[Publications] Ogawa,et al.: "The incrcase of voltage-sensitive calcium channels and calcium accumulation in skeletal muscles of streptozocininduced diabetic rats." Metabolism. (in press).
[Publications] Maegawa et al.: "Thiazolidine derivatives ameliorate glucose induced insulin resistance via the normalization of protein tyrosine phosphatase activitics." J Biol Chem. (in press).
[Publications] Kashiwagi,A.: "Recent advances endothelial cell dysfunction in disbetes." Churchill Livingstone, 286 (1994)
[Publications] Kashiwagi,A.: "Diabetes" Elsevier Science Publisher, 679-682 (1991)

1994 Fiscal Year Annual Research Report

糖尿病における高インスリン血症と動脈硬化-血管平滑筋細胞を用いた分子生物学的研究

Principal Investigator

繁田 幸男 滋賀医科大学, 医学部, 教授 (60028321)

Research Products

[Publications] Ugi ct al.: "Src homology 2 domains of protcin tyrosinc phosphatasc arc associatcd in vitro with both the insulin receptor and insulin receptor substratc-1 via different phosphotyrosinc motifs." FEBS Lcttcr. 340. 216-220 (1994)

[Publications] Maegawa et al.: "Insulin receptor kinase phosphorylates tyrosine phosphatasc containing Src homology 2 region and modulates its PTPase activity in vitro." Biochem Biophys Res Commun. 199. 780-785 (1994)

[Publications] Yasuda et al.: "Mctabolic effect of PGEI analoguc 01206.aCD on ncrvc Na+-K+-ATPasc activity of rais with streptozocin-induccd diabetes is mcdiated via cAMP:Possible role of cAMP in diabetic neuropathy." Prostaglandins. 47. 367-378 (1994)

[Publications] Haneda et al.: "Intcraction between atrial natriurctic peptide and angiotensin II cultured glomcrular mesangial cells and its disturbancc in diabetes." ExP.Nephrol. 2. 106-106 (1994)

[Publications] Kashiwagi et al.: "Abnormal glutathionc mctabolism and increascd cytotoxicity H202 in human umbilical vein endothclial cells culturcd in high glucose medium." Diabetologia. 37. 264-269 (1994)

[Publications] Ide et al.: "High glucose condition activates protein tyrosine phosphatase and deactivates insulin receptor function in insulinscnsitivc rat fibroblasts." Biochem Biophys Res Commun. 201. 71-77 (1994)

[Publications] Ide et al.: "High Glucose Concentration dcsensitizes insulin action at the levcls of receptor kinase." Endocrine J.42. 1-8 (1995)

[Publications] Kikkawa et al.: "Translocation of protein kinaseCα and in rat glomerular mesangial cells cultured under high glucose conditions." Diabetologia. 37. 838-841 (1994)

[Publications] Shigeta et al.: "Diabetic nephropathy in Japan." Diabetic Research and Clinical Practice. 24. S191-S197 (1994)

[Publications] Hidaka et al.: "Effects of lipo prostaglandin E1 injection on transcutancous oxygen tension in diabetic subjects with periphcral vascular discase." J Japanese Collage of Angiology. 34. 433-437 (1994)

[Publications] Takagi et al: "Insulin receptor-mediated signal transduction and its desensitization in culturcd vascular smooth musele cells." Atherosclerosis. 113. 19-27 (1995)

[Publications] Asahina et al.: "Impaired activation of glucose oxidation and NADPH supply in human endothelial cells exposed to H202 in hith glucose medium" Diabctes. (in press).

[Publications] Hidaka et al.: "Effects of an HMG-CoA reductase inhibitor,pravastatin,and bile sequestering resin,cholestyramine,on plasma plant sterol Levels in hypercholesterolemic subjects." Atherosclerosis Thromb. (in press).

[Publications] Sakamoto et al.: "Prevention of glomerular hyperliltration in rats with streptozotocin-induced diabetes by an atrial natriuretic peptide receptor antagonist." Diabetologia. (in press).

[Publications] Takigawa et al.: "Antibodies against GM,anglioside affect K+and N+currents in isolated rat myelinated nerve fibers" Ann Neurol. (in press).

[Publications] Takagi et al.: "Significance of fructose autoxidation protein oxidation and formation of advanced glycation end product." J Diabetes Compl. (in press).

[Publications] Ogawa,et al.: "The incrcase of voltage-sensitive calcium channels and calcium accumulation in skeletal muscles of streptozocininduced diabetic rats." Metabolism. (in press).

[Publications] Maegawa et al.: "Thiazolidine derivatives ameliorate glucose induced insulin resistance via the normalization of protein tyrosine phosphatase activitics." J Biol Chem. (in press).

[Publications] Kashiwagi,A.: "Recent advances endothelial cell dysfunction in disbetes." Churchill Livingstone, 286 (1994)

[Publications] Kashiwagi,A.: "Diabetes" Elsevier Science Publisher, 679-682 (1991)

繁田幸男滋賀医科大学, 医学部, 教授 (60028321)