1996 Fiscal Year Annual Research Report

糖尿病における動脈硬化症危険因子の血管分子生物学:酸化ストレスと高インスリン血症

Research Project

Project/Area Number	07671127
Research Institution	Shiga University of Medical Science
Principal Investigator	柏木厚典滋賀医科大学, 医学部, 助教授 (20127210)
Keywords	糖尿病 / 動脈硬化症危険因子 / 血管生物学 / 酸化ストレス / 高インスリン血症 / リソホスファチジルコリン / 単球走化性因子 / 血管内皮細胞
Research Abstract	糖尿病患者における動脈硬化症は、非糖尿病者に比べて約10年若年から発症し、諸危険因子(高脂血症、高血圧症、喫煙)を補正しても依然として非糖尿病者に比べて2-3倍高頻度であり、患者の生命予後決定因子となっている。そこで糖尿病に特徴的な動脈硬化危険因子(高血糖、酸化LDL、インスリン抵抗性)がどのような機構で動脈硬化発症・進展に関与しているかを血管分子生物学的方法にて検討することは、今後の予防・治療の戦略を検討する上で重要である。既に我々はこの2年間文部省科学研究費を受け以下の点を明らかにしてきた。1)高血糖条件での酸化ストレスの亢進:高血糖状態では、血管内皮細胞での活性酸素処理系としてのグルタチオンレドックス(GR)サイクルが低下し、活性酸素産生の亢進により酸化ストレスが亢進する(Diabctologia 1994,Diabctcs 1995,Diabctcs 1996)。2)高血糖状態に伴う血管内皮細胞の細胞間接着因子(ICAM-1)発現増加-高血糖状態では、血管内皮細胞における細胞接着因子のうちICAM-1が特異的に増加し単球との相互作用が亢進した(Life Science 1994)。3)酸化LDLとMCP-1遺伝子発現:酸化ストレス状態で産生される酸化LDLにより血管内皮細胞の動脈硬化促進性サイトカインとしてMCP-1mRNAが誘導される(Mctabolism 1996)。この活性は血管内皮細胞を抗酸化剤にて処理することにより完全に正常化し、またその誘導機構として転写因子NF-kBの活性化が関与していることが明かとなった(Diabctologia,in press)。4)インスリン抵抗性機構の解明:インスリンによる細胞増殖の調節は、インスリン受容体チロシンキナーゼとプロテインチロシンホスファターゼ(PTPasc)により調節を受けている。本研究では非受容体型PTPascであるPTP1BとSHP2について検討した。高血糖下におけるインスリン作用異常はPTP1Bの活性化によるインスリン受容体脱燐酸化によった(End.J 1995,BBRC 1994)。更にインスリン感受性増強剤により高血糖下PTPasc異常は改善した(J Biol Chem 1995)。一方、促進性PTPascとしてSHP2が存在し、このPTPascドメインを欠損した変異遺伝子を線維芽細胞に導入したところ、インスリン情報伝達系の障害が認められた(BBRC 1994、FEBS icll 1994,J Biol Chem 1996,BBRC 1996)。5)血管平滑筋細胞におけるインスリン抵抗性と細胞増殖シグナル:培養平滑筋細胞(SMCs)を用いて、インスリン作用を検討した。SMCsのインスリン作用はShc、Ras-MAP系情報伝達は認められず、PB-キナーゼの活性化が主な経路であった。このインスリン情報は慢性高インスリン状態にて脱感作され(Athcrosclcrosis 1995)、アミノ酸輸送系(A system)の活性化に関係した(Circ Res 1996)。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] 1.Maegawa,H.,et al.: "Insulin reccptor kinase phosphorylatcs protein tyrosine phosphatase containing Src homology 2 regions and modulates its PTPasc activity in vitro." Biochem Biophys Res Commun. 199. 780-785 (1994)
[Publications] 2.Ugi,S.,et al.: "Src homology 2 domains of protein tyrosine phosphatase are associated in vitro with both the insulin receptor and insulin receptor substrate-1 via different phosphotyrosine motifs." FEBS Lett. 340. 216-220 (1994)
[Publications] 3.Ide,R.,et al.: "High glucose condition activates protein tyrosine phosphatases and deactivates insulin receptor function in insulin-sensitive rat 1 fibroblasts." Biochem Biophys Res Commun. 201. 71-77 (1994)
[Publications] 4.Kashiwagi A,et al.: "Abnormal glutathione metabolism and increased cytotoxicity caused by H_2O_2 in human umbilical vein endothelial cells cultured in high glucose medium." Diabetologia. 37. 264-269 (1994)
[Publications] 5.Maegawa H,et al.: "Thiazolidine derivatives ameliorate high glucose-induced insulin resistance via the normalization of protein-tyrosine phosphatase activities." J Biol Chem. 270. 1-7 (1995)
[Publications] 6.Ide R,et al.: "High glucose concentrantion desensitizes insulin action at the levels of receptor kinase." Endocine J. 42. 1-8 (1995)
[Publications] 7.Takagi Y,et al.: "Insulin-specific activation of S6 kinase and its desensitization in cultured vascular smooth musele cells." Atheroselerosis. 113. 19-27 (1995)
[Publications] 8.Takagi Y,et al.: "Significance of fructose-induced protein oxidation and formation of advanced glycation end product." J.Diab Compl. 9. 87-91 (1995)
[Publications] 9.Ogawa T,et al.: "The increase of voltage-sensitive calcium channels and calcium accumulation in skeletal muscles of streptozocin-induced diabetic rats." Metabilism. 44. 1455-1461 (1995)
[Publications] 10.Asahina T,et al.: "Impaired activation of glucose oxidation and NADPH supply in human endothelial cells exposed to H2O2 in high glucose medium." Diabetes. 44. 520-526 (1995)
[Publications] 11.Kashiwagi A,et al.: "Glucose metabolism and radical scavenger dysfunction in endothelial cells." Diabetes. 45. 584-586 (1996)
[Publications] 12.Taki H,et al.: "Expression of intercellular adhesion molecules 1 (ICAM-1) via an osmotic effect in human umbilical vein endothelial cells exposed to high glucose medium." Life Science. 58. 1713-1721 (1996)
[Publications] 13.Takahara N,et al.: "Lysophosphatidylcholine stimulates the expression and production of MCP-1 by human vascular endothelial cells." Metabolism. 45. 559-564 (1996)
[Publications] 14.Ugi S,et al.: "Expression of dominant negative mutant SHPTP2 attenuates phosphatidylinositol 3'kinase activity via modulation of phosphorylation of insulin receptor substrate-1." J.Biol Chem. 271. 12595-12602 (1996)
[Publications] 15.Maegawa H,et al.: "SHPTP2 servces adapter protein linking between Janus kinase 2 and insulin receptor substrates." Biochem Biophys Res Commun. 228. 122-127 (1996)
[Publications] 16.Sugimoto T,et al.: "Atrial natriuretic peptide induces the expression of MKP-1,a mitogen-activated protein kinase phosphatase,in glomerular mesangial cells." J Biol Chem. 271. 544-547 (1996)
[Publications] 17.Obata T,et al.: "Insulin signaling and its regulation of system A amino acid uptake in cultured rat vascular smooth muscle cells." Circ Res. 79. 1167-1176 (1996)
[Publications] 18.Takahara N,et al.: "Oxidaized lipoprotcins found in patients with non-insulin-dependent diabetes mallitus stimulatc radical-induced MCP-1 mRNA expression in cultured human endothelial cells." Diabetologia. (in press).

1996 Fiscal Year Annual Research Report

糖尿病における動脈硬化症危険因子の血管分子生物学:酸化ストレスと高インスリン血症

Principal Investigator

柏木 厚典 滋賀医科大学, 医学部, 助教授 (20127210)

Research Products

[Publications] 1.Maegawa,H.,et al.: "Insulin reccptor kinase phosphorylatcs protein tyrosine phosphatase containing Src homology 2 regions and modulates its PTPasc activity in vitro." Biochem Biophys Res Commun. 199. 780-785 (1994)

[Publications] 2.Ugi,S.,et al.: "Src homology 2 domains of protein tyrosine phosphatase are associated in vitro with both the insulin receptor and insulin receptor substrate-1 via different phosphotyrosine motifs." FEBS Lett. 340. 216-220 (1994)

[Publications] 3.Ide,R.,et al.: "High glucose condition activates protein tyrosine phosphatases and deactivates insulin receptor function in insulin-sensitive rat 1 fibroblasts." Biochem Biophys Res Commun. 201. 71-77 (1994)

[Publications] 4.Kashiwagi A,et al.: "Abnormal glutathione metabolism and increased cytotoxicity caused by H_2O_2 in human umbilical vein endothelial cells cultured in high glucose medium." Diabetologia. 37. 264-269 (1994)

[Publications] 5.Maegawa H,et al.: "Thiazolidine derivatives ameliorate high glucose-induced insulin resistance via the normalization of protein-tyrosine phosphatase activities." J Biol Chem. 270. 1-7 (1995)

[Publications] 6.Ide R,et al.: "High glucose concentrantion desensitizes insulin action at the levels of receptor kinase." Endocine J. 42. 1-8 (1995)

[Publications] 7.Takagi Y,et al.: "Insulin-specific activation of S6 kinase and its desensitization in cultured vascular smooth musele cells." Atheroselerosis. 113. 19-27 (1995)

[Publications] 8.Takagi Y,et al.: "Significance of fructose-induced protein oxidation and formation of advanced glycation end product." J.Diab Compl. 9. 87-91 (1995)

[Publications] 9.Ogawa T,et al.: "The increase of voltage-sensitive calcium channels and calcium accumulation in skeletal muscles of streptozocin-induced diabetic rats." Metabilism. 44. 1455-1461 (1995)

[Publications] 10.Asahina T,et al.: "Impaired activation of glucose oxidation and NADPH supply in human endothelial cells exposed to H2O2 in high glucose medium." Diabetes. 44. 520-526 (1995)

[Publications] 11.Kashiwagi A,et al.: "Glucose metabolism and radical scavenger dysfunction in endothelial cells." Diabetes. 45. 584-586 (1996)

[Publications] 12.Taki H,et al.: "Expression of intercellular adhesion molecules 1 (ICAM-1) via an osmotic effect in human umbilical vein endothelial cells exposed to high glucose medium." Life Science. 58. 1713-1721 (1996)

[Publications] 13.Takahara N,et al.: "Lysophosphatidylcholine stimulates the expression and production of MCP-1 by human vascular endothelial cells." Metabolism. 45. 559-564 (1996)

[Publications] 14.Ugi S,et al.: "Expression of dominant negative mutant SHPTP2 attenuates phosphatidylinositol 3'kinase activity via modulation of phosphorylation of insulin receptor substrate-1." J.Biol Chem. 271. 12595-12602 (1996)

[Publications] 15.Maegawa H,et al.: "SHPTP2 servces adapter protein linking between Janus kinase 2 and insulin receptor substrates." Biochem Biophys Res Commun. 228. 122-127 (1996)

[Publications] 16.Sugimoto T,et al.: "Atrial natriuretic peptide induces the expression of MKP-1,a mitogen-activated protein kinase phosphatase,in glomerular mesangial cells." J Biol Chem. 271. 544-547 (1996)

[Publications] 17.Obata T,et al.: "Insulin signaling and its regulation of system A amino acid uptake in cultured rat vascular smooth muscle cells." Circ Res. 79. 1167-1176 (1996)

[Publications] 18.Takahara N,et al.: "Oxidaized lipoprotcins found in patients with non-insulin-dependent diabetes mallitus stimulatc radical-induced MCP-1 mRNA expression in cultured human endothelial cells." Diabetologia. (in press).

柏木厚典滋賀医科大学, 医学部, 助教授 (20127210)