1997 Fiscal Year Annual Research Report

cGMP系の血管平滑筋細胞分化決定における新しい意義に関する分子医学的研究

Research Project

Project/Area Number	09281219
Research Institution	Kyoto University
Principal Investigator	伊藤裕京都大学, 医学研究科, 助手 (40252457)
Keywords	血管平滑筋細胞 / 増殖 / cGMP / ナトリウム利尿ペプチド / ホメオボックス / キナーゼ / 細胞周期
Research Abstract	これまで我々は、ナトリウム利尿ペプチドがcGMP系を活性化しvascular growthに対して抑制的に作用することを明らかにした。更に、ナトリウム利尿ペプチドの一つであるCNPが血管内皮細胞において生合成されることを明らかにし、CNPがNOと並ぶペプチド性のEDRFであることを証明し、“血管壁ナトリウム利尿ペプチド系"という概念を提唱した。最近、CNP遺伝子組み換えアデノウイルスを構築し、培養血管平滑筋細胞にCNP遺伝子を導入し過剰発現させることでcGMP系を活性化すると、増殖がGl期で停止するとともに、再分化が生ずるという新しいcGMP系の作用を報告した。更に、心血管系特異的ホメオボックス遺伝子であるGaxがcGMP系の活性化によりその遺伝子発現が亢進することも報告し、cGMP系において転写因子Gaxという新しいシグナル伝達経路が存在する可能性を提示した。本年度は、cGMPカスケードにおいてcGMP産生よりGax発現亢進に至るシグナル伝達経路を明らかにするため、我々が既にクローニングに成功したヒトcGMP依存性プロティンキナーゼ(Gキナーゼ)タイプIαについて検討した。すなわち、GキナーゼのN端部の二重化ドメイン、cGMP結合ドメインを欠失させたミュータントを作成し、βアクチンプロモーター、サイトメガロウィルスエンハンサーを有する発現ベクターを構築した。Gキナーゼ特異的基質を用いたin vitroのGキナーゼkination assayを確立し、このミュータントGキナーゼがcGMPを必要としない持続活性型constitutive activeであることを明らかにした。更に、このミュータントGキナーゼ発現ベクターを用い、培養血管平滑筋細胞の遊走、増殖が著明に抑制されることを明らかにし、cGMPカスケードによる血管平滑筋細胞のフェノタイプ決定におけるGキナーゼの関与を証明した。更に、CNP-cGMP-Gキナーゼ-Gaxに至る血管平滑筋分化決定におけるシグナル伝達経路においてcyclin/CDK complexの抑制因子であるp21^<CIPl>の関与を明らかにした。

Research Products

(17 results)

All Other

All Publications (17 results)

[Publications] J.Yamashita, et al.: "Opposite regulation of Gax homeobox expression by angiotensin II and C-type natriuretic peptide." Hypertension. 29. 381-387 (1997)
[Publications] T-H.Chun, et al.: "Shear stress augments expression of C-type natriuretic peptide and adrenomedullin." Hypertension. 29. 1296-1302 (1997)
[Publications] T.Ikeda, et al.: "The plasma levels of brain and atrial natriuretic peptides elevate in proportion to the left ventricular end-systolic wall stress in patients with aortic stenosis." Am.Heart J.133. 307-314 (1997)
[Publications] H.Itoh, et al.: "Impaired guanosine 3',5'-cyclin phosphate production in severe pregnancy-induced hypertension with high plasma levels of atrial and brain natriuretic pepitdes." Endocrine J.44. 389-393 (1997)
[Publications] K.Yoshimine, et al: "Altered expression of atrial natriuretic peptide and contractile protein genes in hypertrophied ventricle of JVS mice with systemic carnitine deficiency." J.Mol.Cell Cardiol.29. 571-578 (1997)
[Publications] H.Itoh, et al.: "Significance of vascular natriuretic peptide system in vascular remodeling in humans and its application to gene therapy." Annals New York Academy of Sciences. 811. 533-541 (1997)
[Publications] M.Harada, et al.: "Significance of ventricular myocytes and nonmyocytes interaction during cardiocyte hypertrophy-Evidence for endothelin-1 as a paracrine hypertrophic factor from cardiac nonmyocytes." Circulation. 96. 3737-3744 (1997)
[Publications] K.Doi, et al: "Adenovirus-mediated gene transfer of C-type natriuretic peptide causes Gl growth inhibition of cultured vascular smooth muscle cells." Biochem.Biophys.Res.Commun.239. 889-894 (1997)
[Publications] J.Matsuda.et al: "Cloning of rat uncoupling protein-3 and uncoupling protein-2 cDNAs : their gene expression in rats fed high-fat diet." FEBS Letters. 418. 200-204 (1997)
[Publications] K.Doi, et al: "Expression of natriuretic peptide system during embryonic stem cell vasculogenesis." Heart and Vessels. Suppl.12. 18-22 (1997)
[Publications] M.Harada, et al.: "Role of cardiac nonmyocytes in cyclic mechanical stretch-induced myocyte hypertrophy." Heart and Vessels. Suppl.12. 198-200 (1997)
[Publications] K.Masatsugu, et al: "Physiologic shear stress suppresses endothelin-converting enzyme-l 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] S.Suga, et al.: "Regulation of endothelial production of C-type natriuretic peptide by interaction between endothelial cells and macrophages." Endocrinology. (in press). (1998)
[Publications] M.Suda, et al.: "Skeletal overgrowth in transgenic mice that overexpress brain natriuretic peptide." Proc.Natl.Acad.Sci.U.S.A. (in press). (1998)
[Publications] Y.Numata, et al.: "An immunoradiometric assay kit for N-terminal fragment of proatrial natriuretic peptide (Pro ANP N-peptide) in human plasma as a useful clinical test for heart failure." Endocrinol.Metabol.(in press). (1998)
[Publications] H.Itoh, et al.ed.by W.K.Samson and E.R.Levin: "“Contemporary Endocrinology : Natriuretic Peptides in Health and Disease"" Humana Press Inc.,Totowa,NJ, (1997)

1997 Fiscal Year Annual Research Report

cGMP系の血管平滑筋細胞分化決定における新しい意義に関する分子医学的研究

Principal Investigator

伊藤 裕 京都大学, 医学研究科, 助手 (40252457)

Research Products

[Publications] J.Yamashita, et al.: "Opposite regulation of Gax homeobox expression by angiotensin II and C-type natriuretic peptide." Hypertension. 29. 381-387 (1997)

[Publications] T-H.Chun, et al.: "Shear stress augments expression of C-type natriuretic peptide and adrenomedullin." Hypertension. 29. 1296-1302 (1997)

[Publications] T.Ikeda, et al.: "The plasma levels of brain and atrial natriuretic peptides elevate in proportion to the left ventricular end-systolic wall stress in patients with aortic stenosis." Am.Heart J.133. 307-314 (1997)

[Publications] H.Itoh, et al.: "Impaired guanosine 3',5'-cyclin phosphate production in severe pregnancy-induced hypertension with high plasma levels of atrial and brain natriuretic pepitdes." Endocrine J.44. 389-393 (1997)

[Publications] K.Yoshimine, et al: "Altered expression of atrial natriuretic peptide and contractile protein genes in hypertrophied ventricle of JVS mice with systemic carnitine deficiency." J.Mol.Cell Cardiol.29. 571-578 (1997)

[Publications] H.Itoh, et al.: "Significance of vascular natriuretic peptide system in vascular remodeling in humans and its application to gene therapy." Annals New York Academy of Sciences. 811. 533-541 (1997)

[Publications] M.Harada, et al.: "Significance of ventricular myocytes and nonmyocytes interaction during cardiocyte hypertrophy-Evidence for endothelin-1 as a paracrine hypertrophic factor from cardiac nonmyocytes." Circulation. 96. 3737-3744 (1997)

[Publications] K.Doi, et al: "Adenovirus-mediated gene transfer of C-type natriuretic peptide causes Gl growth inhibition of cultured vascular smooth muscle cells." Biochem.Biophys.Res.Commun.239. 889-894 (1997)

[Publications] J.Matsuda.et al: "Cloning of rat uncoupling protein-3 and uncoupling protein-2 cDNAs : their gene expression in rats fed high-fat diet." FEBS Letters. 418. 200-204 (1997)

[Publications] K.Doi, et al: "Expression of natriuretic peptide system during embryonic stem cell vasculogenesis." Heart and Vessels. Suppl.12. 18-22 (1997)

[Publications] M.Harada, et al.: "Role of cardiac nonmyocytes in cyclic mechanical stretch-induced myocyte hypertrophy." Heart and Vessels. Suppl.12. 198-200 (1997)

[Publications] K.Masatsugu, et al: "Physiologic shear stress suppresses endothelin-converting enzyme-l 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] S.Suga, et al.: "Regulation of endothelial production of C-type natriuretic peptide by interaction between endothelial cells and macrophages." Endocrinology. (in press). (1998)

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

[Publications] Y.Numata, et al.: "An immunoradiometric assay kit for N-terminal fragment of proatrial natriuretic peptide (Pro ANP N-peptide) in human plasma as a useful clinical test for heart failure." Endocrinol.Metabol.(in press). (1998)

[Publications] H.Itoh, et al.ed.by W.K.Samson and E.R.Levin: "“Contemporary Endocrinology : Natriuretic Peptides in Health and Disease"" Humana Press Inc.,Totowa,NJ, (1997)

伊藤裕京都大学, 医学研究科, 助手 (40252457)