新規血管虚血負荷装置の開発と新しい虚血シグナルの同定および虚血血管再生遺伝子治療

• Project/Area Number: 09877192
• Research Category: Grant-in-Aid for Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: 内分泌・代謝学
• Research Institution: Kyoto University
• Principal Investigator: 伊藤 裕 京都大学, 医学研究科, 助手 (40252457)
• Project Period (FY): 1997 – 1998
• Project Status: Completed (Fiscal Year 1998)
• Budget Amount: ¥1,900,000 (Direct Cost: ¥1,900,000); Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000); Fiscal Year 1997: ¥1,300,000 (Direct Cost: ¥1,300,000)
• Keywords: 内皮細胞 / 虚血 / ずり応力 / 酸化ストレス / 血管作動性物質 / 血管新生 / VEGF

Research Abstract

本研究は、新しい血管虚血負荷装置の開発と、それを用いた内皮細胞での虚血応答遺伝子発現プログラムを明らかにし、その知見を虚血血管再生遺伝子治療に応用することをその目的とする。昨年度は、内皮細胞において虚血刺激によりその遺伝子発現が誘導されることが知られている血管新生因子VEGF(VascularEndothelial Growth Factor)に注目し、その遺伝子発現のシグナル機構を低酸素チャンバーを用い検討した。その結果、虚血シグナルを考える上で、酸化LDLやLysoPCのシグナル伝達に関与する経路の重要性を明らかにした。本年度は、虚血刺激のもう一つの重要なコンポーネントであるずり応力(shear stress)の低下について、平行平板型ずり応力負荷装置を用い検討した。ずり応力の増加により、血管拡張ペプチドであるC型ナトリウム利尿ペプチド(CNP)及びアドレノメジュリン(AM)の発現が亢進し、一方血管収縮ペプチドであるエンドセリン(ET)及びその産生に関与するエンドセリン変換酵素(ECE)の発現は低下した。これらのずり応力の内皮由来血管作動性ペプチド発現制御は、酸化ストレスそのものであるH202によりmimicされた。更に、ずり応力を低下させることにより、ET、ECE発現が上昇することも明らかにした。従って、虚血シグナルを考える上で、血流の減少によるずり応力の低下に伴う酸化ストレスの変化が重要であることが明かとなった。更に、ずり応力の低下によりその発現の低下するCNPのアデノウィルスを用いた血管への遺伝子導入により、血管平滑筋細胞の増殖抑制・分化誘導及び内皮細胞の再生の促進が起こることが明かとなり、CNPの虚血血管再生遺伝子治療への応用の有用性が示唆された。

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 natriuretic 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.: "Vascular endothelial growth factoer(VEGF)expression in human coronary atherosclerotic lesions:Possible pathophysiological signifiance 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 on 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 natriuretic 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 inhibition 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." Molecular 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)
• [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 facter from cardiac nonmyocytes." Circulation. 96. 3737-3744 (1997)
• [Publications] K.Doi, et al.: "Adenovirus-mediated gene transfer of C-type natriuretic peptide causes G1 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.Masatsuge, et al.: "Physiologic shear stress suppresses endothelin-converting enzyme-1 expression in vascular endothelial cells." J.Cardiovasc.Pharmacol.31Suppl.1. S42-S45 (1998)
• [Publications] T.Saito, et al.: "Oxidative stress suppresses the endothelial secretion of endothelin." J.Cardiovasc.Pharmacol.31Suppl.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)