1998 Fiscal Year Annual Research Report

内皮分化における血管局所ホルモンの意義に関する発生学的検討と血管再生治療の試み

Research Project

Project/Area Number	10470227
Research Institution	Kyoto University
Principal Investigator	伊藤裕京都大学, 医学研究科, 助手 (40252457)
Co-Investigator(Kenkyū-buntansha)	細田公則京都大学, 人間環境学研究科, 助手 (40271598) 梅村和夫浜松医科大学, 助教授 (40232912) 小川峰太郎京都大学, 医学研究科, 助手 (70194454)
Keywords	内皮細胞 / 血管平滑筋細胞 / 前駆細胞 / ES細胞 / 動脈硬化症 / VEGF / cGMP / トランスジェニックマウス
Research Abstract	増殖性血管病変は、内皮細胞の機能破綻とそれに基づく血管平滑筋細胞の脱分化による遊走・増殖等によりもたらされる。従って、内皮細胞の機能分化の分子レベルでの解明は、内皮機能異常のメカニズムの理解に直結し、更に血管機能再生治療への応用につながる。そこで本研究では、我々がこれまで一貫して研究してきたナトリウム利尿ペプチド系を中心に血管局所ホルモンに焦点を当て、その内皮再生分化における病態生理的意義に関し、特に発生学的手法を駆使して検討することを目的とする。本年度は以下の研究結果を得た。 (1) ヒト冠動脈硬化病変における血管内皮細胞増殖因子(VEGF)及びその受容体(flk-1,flt-1)の発現の検討: ヒト冠動脈硬化病変において、内皮細胞でのVEGF発現が亢進していること、及び脱分化した血管平滑筋細胞及び病変部に集積するマクロファージにおいて、VEGF,flk-1,flt-1発現が認められることを明らかにした。 (2) フローサイトメーターによるマウスES細胞からの内皮細胞前駆細胞のソーティングと内皮細胞分化における血管局所ホルモンの意義の検討:マウスES細胞より、VEGF受容体flk-1陽性細胞をFACSアナライザーによりソーティングし、この細胞にVEGFを添加することによりVEカドヘリン陽性の内皮細胞の誘導に成功した。更にこの細胞を用い、コラーゲンゲル三次元培養にて管腔構造の構成に成功した。 (3) cGMP依存性プロテインキナーゼ(cGK)トランスジェニックマウスの開発と内皮損傷、血栓形成及び管平滑筋増殖モデルでの検討:サイトメガロウイルスエンハンサー、βアクチンプロモーターにヒトcGKIαcDNAを連結したトランスジーンを用いたcGK過剰発現トランスジェニックマウス(cGK.Tg)を開発した。cGK遺伝子を3コピー及び15コピー有する2系統のTgが得られ、これらにおいて心臓、肺、血管、骨格筋、小脳において高いcGK活性が認められた。内皮損傷、血栓形成、新生内膜増殖モデルにおいて、cGK.Tgでは、血管平滑筋細胞遊走・増殖抑制傾向が認められた。

Research Products
(19 results)

All Other

All Publications (19 results)

[Publications] K.Masatsugu et al.: "Physiologic shear stress suppresses endothelin-converting enzyme-1 expression in vascular endothelial cells." J.Cardiovasc.Pharmacol.31 SUPPL 1. S42-S45 (1998)
[Publications] T.Saito et al.: "Oxidative stress suppresses the endothelial secretion of endothelin." J.Cardiovasc.Pharmacol.31 SUPPL 1. S45-S47 (1998)
[Publications] T.Igaki et al.: "Effects of intravenously administered C-type natruretic peptide in humans--Comparison with atrial natriuretic peptide." Hypertens.Res.21. 7-13 (1998)
[Publications] S.Suga et al.: "Regulation of endothelial production of C-type natriuretic peptide by interaction between endothelial cells and macrophages." Endocrinology. 139. 1920-1926 (1998)
[Publications] M.Suda et al.: "Skeletal overgrowth in transgenic mice that overexpress brain natriuretic peptide." Proc.Natl.Acad.Sci.U.S.A.95. 2337-2342 (1998)
[Publications] N.Azuma et al.: "The significance of the Trp 64 Arg mutation of the b3-adrenergic receptor gene in impaired glucose tolerance, non-insulin-dependent diabetes mellitus, and insulin resistance in Japanese subjects." Metabolism. 47. 456-460 (1998)
[Publications] M.Inoue et al.: "Vacular endothelial growth factor(VEGF)expression in human coronary atherosclerotic lesions:Possible pathophysiological significance of VEGF in progression of atherosclerosis." Circulation. 98. 2108-2116 (1998)
[Publications] Y.Miyamoto et al.: "Endothelial nitric oxide synthase gene is positively associated with essential hypertension." Hypertension. 32. 3-8 (1998)
[Publications] J.Matsuda et al.: "Increased adipose expression of uncoupling protein-3 gnene by thiazolidinediones in Wistar fatty rats and in cultured adipocytes." Diabetes. 47. 1809-1814 (1998)
[Publications] M.Yamaguchi et al.: "Effect of neutral endopeptidase inhibition of the natriuresis and renal clearance of atrial natriuretic peptide in perfused rat kidney." Pharmaceutical Research. 15. 1499-1502 (1998)
[Publications] K.Doyama et al.: "Expression and distribution of brain natruiretic peptide in human right atria." J.Am.Coll.Cardiol.32. 1832-1838 (1998)
[Publications] H.Itoh et al.: "Hypertension and insulin resistance-The role of peroxisome proliferator-activated receptor-γ(PPARγ)-" Clinical and Experimental Pharmacology and Physiology. (in press). (1999)
[Publications] Y.Numata et al.: "Immunoradiometric assay for the N-terminal fragment of proatrial natriuretic peptide in human plasma." Clinical Chemistry. 44. 1008-1013 (1998)
[Publications] M.Harada et al.: "Interaction of myocytes and nonmyocytes is necessary for mechanical stretch to induce ANP/BNP production in cardiocyte culture." J.Cardiovasc.Pharmacol.31 SUPPL 1. S357-S359 (1998)
[Publications] G.Takemura et al.: "Expression of atrial and brain natriuretic peptides and their genes in hearts of patients with cardiac amyloidosis." J.Am.Coll.Cardiol.31. 254-265 (1998)
[Publications] J.Yamashita et al.: "Identification of cis-element of the human endothelin-A receptor gene and inhobition of the gene expression by the decoy strategy." J.Biol.Chem.273. 15993-15999 (1998)
[Publications] H.Nanno et al.: "Plasma nitric oxide metabolite levels are decreased in pre-eclamptic women complicated with fetal distress." Prenat.Neonat Med.3. 222-226 (1998)
[Publications] H.Nanno et al.: "Nitric oxide metabolite concentrations in maternal plasma decrease during parturition: possible transient down-regulation of nitric oxide synthesis." Molecualr Human Reproduction. 4. 609-616 (1998)
[Publications] T.Tanaka et al.: "Down-regulation of peroxisome proliferator-activated receptorg expression by inflammatory cytokines and its reversal by thiazolidinediones." Diabetologia. (in press). (1999)

1998 Fiscal Year Annual Research Report

内皮分化における血管局所ホルモンの意義に関する発生学的検討と血管再生治療の試み

Principal Investigator

伊藤 裕 京都大学, 医学研究科, 助手 (40252457)

Research Products

[Publications] K.Masatsugu et al.: "Physiologic shear stress suppresses endothelin-converting enzyme-1 expression in vascular endothelial cells." J.Cardiovasc.Pharmacol.31 SUPPL 1. S42-S45 (1998)

[Publications] T.Saito et al.: "Oxidative stress suppresses the endothelial secretion of endothelin." J.Cardiovasc.Pharmacol.31 SUPPL 1. S45-S47 (1998)

[Publications] T.Igaki et al.: "Effects of intravenously administered C-type natruretic peptide in humans--Comparison with atrial natriuretic peptide." Hypertens.Res.21. 7-13 (1998)

[Publications] S.Suga et al.: "Regulation of endothelial production of C-type natriuretic peptide by interaction between endothelial cells and macrophages." Endocrinology. 139. 1920-1926 (1998)

[Publications] M.Suda et al.: "Skeletal overgrowth in transgenic mice that overexpress brain natriuretic peptide." Proc.Natl.Acad.Sci.U.S.A.95. 2337-2342 (1998)

[Publications] N.Azuma et al.: "The significance of the Trp 64 Arg mutation of the b3-adrenergic receptor gene in impaired glucose tolerance, non-insulin-dependent diabetes mellitus, and insulin resistance in Japanese subjects." Metabolism. 47. 456-460 (1998)

[Publications] M.Inoue et al.: "Vacular endothelial growth factor(VEGF)expression in human coronary atherosclerotic lesions:Possible pathophysiological significance of VEGF in progression of atherosclerosis." Circulation. 98. 2108-2116 (1998)

[Publications] Y.Miyamoto et al.: "Endothelial nitric oxide synthase gene is positively associated with essential hypertension." Hypertension. 32. 3-8 (1998)

[Publications] J.Matsuda et al.: "Increased adipose expression of uncoupling protein-3 gnene by thiazolidinediones in Wistar fatty rats and in cultured adipocytes." Diabetes. 47. 1809-1814 (1998)

[Publications] M.Yamaguchi et al.: "Effect of neutral endopeptidase inhibition of the natriuresis and renal clearance of atrial natriuretic peptide in perfused rat kidney." Pharmaceutical Research. 15. 1499-1502 (1998)

[Publications] K.Doyama et al.: "Expression and distribution of brain natruiretic peptide in human right atria." J.Am.Coll.Cardiol.32. 1832-1838 (1998)

[Publications] H.Itoh et al.: "Hypertension and insulin resistance-The role of peroxisome proliferator-activated receptor-γ(PPARγ)-" Clinical and Experimental Pharmacology and Physiology. (in press). (1999)

[Publications] Y.Numata et al.: "Immunoradiometric assay for the N-terminal fragment of proatrial natriuretic peptide in human plasma." Clinical Chemistry. 44. 1008-1013 (1998)

[Publications] M.Harada et al.: "Interaction of myocytes and nonmyocytes is necessary for mechanical stretch to induce ANP/BNP production in cardiocyte culture." J.Cardiovasc.Pharmacol.31 SUPPL 1. S357-S359 (1998)

[Publications] G.Takemura et al.: "Expression of atrial and brain natriuretic peptides and their genes in hearts of patients with cardiac amyloidosis." J.Am.Coll.Cardiol.31. 254-265 (1998)

[Publications] J.Yamashita et al.: "Identification of cis-element of the human endothelin-A receptor gene and inhobition of the gene expression by the decoy strategy." J.Biol.Chem.273. 15993-15999 (1998)

[Publications] H.Nanno et al.: "Plasma nitric oxide metabolite levels are decreased in pre-eclamptic women complicated with fetal distress." Prenat.Neonat Med.3. 222-226 (1998)

[Publications] H.Nanno et al.: "Nitric oxide metabolite concentrations in maternal plasma decrease during parturition: possible transient down-regulation of nitric oxide synthesis." Molecualr Human Reproduction. 4. 609-616 (1998)

[Publications] T.Tanaka et al.: "Down-regulation of peroxisome proliferator-activated receptorg expression by inflammatory cytokines and its reversal by thiazolidinediones." Diabetologia. (in press). (1999)

伊藤裕京都大学, 医学研究科, 助手 (40252457)