1998 Fiscal Year Annual Research Report

メイラード反応後期生成物(AGE)に対する生体の除去機能-インスリンシグナルによるAGEレセプターの機能制御-

Research Project

Project/Area Number	09470225
Research Institution	Kumamoto University
Principal Investigator	堀内正公熊本大学, 医学部, 教授 (10117377)
Co-Investigator(Kenkyū-buntansha)	荒木栄一熊本大学, 医学部付属病院, 講師 (10253733)
Keywords	メイラード後期生成物(AGE) / 糖尿病合併症 / マクロファージ / スカベンジャー受容体 / インスリン / lyn / MAP kinase / 細胞内情報伝達系
Research Abstract	蛋白はグルコースによって非酵素的糖化反応を受けて後期反応生成物(AGE)を形成する。生体蛋白のAGE化は高血糖状態や酸化ストレスによって促進され、糖尿病合併症や動脈硬化症などの病因として注目されている。AGEは細胞膜上のAGE受容体を介して種々の細胞現象を誘発する。我々は細胞レベルの解析から、MSR-A(Macrophage Scavenger Receptor typeA)がマクロファージの主要AGE受容体であることを明確にした。本研究の初年度、我々はMSR-Aを介するAGE蛋白の取り込み・分解がインスリンによって促進する現象を見い出し、これにはインスリン受容体を介してIRS-1からPI3 kinaseに続く細胞内情報伝達系が関与していることを明らかにした。更にMSR-Aと細胞内情報伝達系の機能的連関を明らかにするため、平成11年度はMSR-Aと細胞内で会合する蛋白として細胞質チロシンキナーゼであるlynに注目した。このため、我々はMSR-AのリガンドであるアセチルLDLによってマクロファージ増殖が誘発され、この時、MAP kinaseの活性化を伴う事実を利用した。先ず、lyn抗体及びMSR-A抗体を用いた免疫沈降法によって、マクロファージ系細胞であるRAW264.7細胞ではlynとMSR-Aが会合していることが明らかになった。更に、MSR-AとlynをCHO細胞に共発現させたCHO-MSR-lyn細胞とlynの代わりにキナーゼ活性を欠損させたlyn(kinase domainの397番目のリジンをメチオニンに置換)を共発現させたCHO-MSR-lyn(K397M)細胞を調製した。CHO-MSR-lyn細胞をアセチルLDLと37度で5分間保温すると、MAP kinaseのリン酸化反応がコントロールの5倍に上昇した。これに対して、lyn kinaseを欠くCHO-MSR-lyn(K397M)細胞のMAP kinaseのリン酸化反応は有意に上昇しなかった。これらの知見はlynのリン酸化がMSR-AによるMAP kinaseの活性化機構に重要な役割を担っていることを示唆している。

Research Products

(18 results)

All Other

All Publications (18 results)

[Publications] Ryoji Nagai et al.: "Conversion of Amadori product of Maillard reaction to N ε-(carboxymethyl)lysine in alkaline conditions" FEBS Letts.425. 355-360 (1998)
[Publications] Kasuyoshi Ikeda et al.: "Immunochemical approaches to AGE-structures:characterization of anti-AGE antibodies" J.Immunol.Methods. 215. 95-104 (1998)
[Publications] Hiroyuki Sano et al.: "Insulin enhances macrophage scavenger receptor-mediated endocytic uptake of advanced glycation end products" J.Biol.Chem.273. 8630-8637 (1998)
[Publications] Noriyuki Sakata et al.: "Increased advanced glycation end products in atheroscleroric lesions of patients with end-stage renal disease" Atherosclerosis. 142. 67-77 (1999)
[Publications] Yoshiteru Jinnouchi et al.: "Clycolaldehyde-modified low density lipoprotein leads macrophages to foam cells via the macrophage scavenger receptor" J.Biochem.123. 1208-1217 (1998)
[Publications] Yoshida,S. et al.: "Immunohistochemical study of human advanced glycation end-products(AGE)and growth factors in cardiac tissues of patients on maintenance dialysis and with kidney trasplantation" Clin.Nephrol.49. 273-280 (1998)
[Publications] Jing Meng et al.: "Glycoxidation in aortic collagen from STZ-induced diabetic rats and its relevance to vascular damage" Atherosclerosis. 136. 355-365 (1998)
[Publications] Kazuyoshi Ikeda et al.: "Determination of glycated albumin by enzyme-linked boronate-immunoassay(ELBIA)" Clin.Chem.442. 256-263 (1998)
[Publications] Hideki Hakamata et al.: "Isolation of macrophage-like cell mutants resistant to the cytotoxic effect of oxidized low density lipoprotein" J.Lipid Res.39. 482-494 (1998)
[Publications] Hirofumi Matsuda et al.: "Molecular cloning,functional expression,tissue distribution and activation by acetylated LDL in macrophages of rat acyl-coenzyme A:cholesterol acyltransferase" Biochem.Biophys.Acta.1391. 193-203 (1998)
[Publications] Yi Ding et al.: "Reduced expression of the macrophage scavenger receptors in macrophage-like cell mutants resistant to Brefeldin A." Biochem.Biophys.Res.Commun.243. 277-283 (1998)
[Publications] Hisashi Sakaguchi et al.: "Role of macrophage scavenger receptors in fat-induced atherosclerosis in mice" Lab.Invest.78. 423-434 (1998)
[Publications] Takemi Kimura et al.: "Localization of identified advanced glycation end product structures,N ε-(carboxymethyl)-lysine and pentosidine,in age-related inclusions in human brains" Pathol.Intenational.48. 575-579 (1998)
[Publications] Ken Matsuse et al.: "Immunohistochemical detection of advanced glycation end products in pulmonary fibrosis" J.Clin.Pathol.51. 515-519 (1998)
[Publications] Tatsuro Ishibashi et al.: "Nε-(Carboxymethyl)lysine accumulates in soft drusen and choroidal neovascular membranes" Arch.Opthalmol.116. 1629-1632 (1998)
[Publications] Noriyuki Sakata et al.: "Immunohistochemical localization of different epitopes of advanced glycation end products in human atherosclerotic lesions" Atherosclerosis. 141. 61-75 (1998)
[Publications] Ling,X.et al.: "Immunohistochemical distribution and subcellular localization of three distinct specific molecular structures of advanced glycation end products in human tissues" Lab.Invest.78. 1591-1606 (1998)
[Publications] Masahiko Kushiro et al.: "Accumulation of N-episilon-(carbox ymethyllysine)and changes in glomerular extracellular matrix components in Otsuka Long-Evans Tokushima fatty rat:a model of sponteneous NIDDM" Nehpron. 79. 458-468 (1998)

1998 Fiscal Year Annual Research Report

メイラード反応後期生成物(AGE)に対する生体の除去機能-インスリンシグナルによるAGEレセプターの機能制御-

Principal Investigator

堀内 正公 熊本大学, 医学部, 教授 (10117377)

Research Products

[Publications] Ryoji Nagai et al.: "Conversion of Amadori product of Maillard reaction to N ε-(carboxymethyl)lysine in alkaline conditions" FEBS Letts.425. 355-360 (1998)

[Publications] Kasuyoshi Ikeda et al.: "Immunochemical approaches to AGE-structures:characterization of anti-AGE antibodies" J.Immunol.Methods. 215. 95-104 (1998)

[Publications] Hiroyuki Sano et al.: "Insulin enhances macrophage scavenger receptor-mediated endocytic uptake of advanced glycation end products" J.Biol.Chem.273. 8630-8637 (1998)

[Publications] Noriyuki Sakata et al.: "Increased advanced glycation end products in atheroscleroric lesions of patients with end-stage renal disease" Atherosclerosis. 142. 67-77 (1999)

[Publications] Yoshiteru Jinnouchi et al.: "Clycolaldehyde-modified low density lipoprotein leads macrophages to foam cells via the macrophage scavenger receptor" J.Biochem.123. 1208-1217 (1998)

[Publications] Yoshida,S. et al.: "Immunohistochemical study of human advanced glycation end-products(AGE)and growth factors in cardiac tissues of patients on maintenance dialysis and with kidney trasplantation" Clin.Nephrol.49. 273-280 (1998)

[Publications] Jing Meng et al.: "Glycoxidation in aortic collagen from STZ-induced diabetic rats and its relevance to vascular damage" Atherosclerosis. 136. 355-365 (1998)

[Publications] Kazuyoshi Ikeda et al.: "Determination of glycated albumin by enzyme-linked boronate-immunoassay(ELBIA)" Clin.Chem.442. 256-263 (1998)

[Publications] Hideki Hakamata et al.: "Isolation of macrophage-like cell mutants resistant to the cytotoxic effect of oxidized low density lipoprotein" J.Lipid Res.39. 482-494 (1998)

[Publications] Hirofumi Matsuda et al.: "Molecular cloning,functional expression,tissue distribution and activation by acetylated LDL in macrophages of rat acyl-coenzyme A:cholesterol acyltransferase" Biochem.Biophys.Acta.1391. 193-203 (1998)

[Publications] Yi Ding et al.: "Reduced expression of the macrophage scavenger receptors in macrophage-like cell mutants resistant to Brefeldin A." Biochem.Biophys.Res.Commun.243. 277-283 (1998)

[Publications] Hisashi Sakaguchi et al.: "Role of macrophage scavenger receptors in fat-induced atherosclerosis in mice" Lab.Invest.78. 423-434 (1998)

[Publications] Takemi Kimura et al.: "Localization of identified advanced glycation end product structures,N ε-(carboxymethyl)-lysine and pentosidine,in age-related inclusions in human brains" Pathol.Intenational.48. 575-579 (1998)

[Publications] Ken Matsuse et al.: "Immunohistochemical detection of advanced glycation end products in pulmonary fibrosis" J.Clin.Pathol.51. 515-519 (1998)

[Publications] Tatsuro Ishibashi et al.: "Nε-(Carboxymethyl)lysine accumulates in soft drusen and choroidal neovascular membranes" Arch.Opthalmol.116. 1629-1632 (1998)

[Publications] Noriyuki Sakata et al.: "Immunohistochemical localization of different epitopes of advanced glycation end products in human atherosclerotic lesions" Atherosclerosis. 141. 61-75 (1998)

[Publications] Ling,X.et al.: "Immunohistochemical distribution and subcellular localization of three distinct specific molecular structures of advanced glycation end products in human tissues" Lab.Invest.78. 1591-1606 (1998)

[Publications] Masahiko Kushiro et al.: "Accumulation of N-episilon-(carbox ymethyllysine)and changes in glomerular extracellular matrix components in Otsuka Long-Evans Tokushima fatty rat:a model of sponteneous NIDDM" Nehpron. 79. 458-468 (1998)

堀内正公熊本大学, 医学部, 教授 (10117377)