1996 年度実績報告書

血管内皮・平滑筋細胞機能を細胞分子レベルで生体連続観察できるシステムの開発

研究課題

研究課題/領域番号	06557045
研究機関	九州大学
研究代表者	金出英夫九州大学, 医学部, 教授 (80038851)
研究分担者	西村淳二九州大学, 医学部, 助教授 (90237727) 小林誠九州大学, 医学部, 助教授 (80225515)
キーワード	血管内皮 / 血管平滑筋 / 生体計測 / 細胞質カルシウム / フラ-2
研究概要	(1)昨年完成した「血管内皮・平滑筋細胞機能を細胞分子レベルで生体連続観察できるシステム・第2号機」は細胞内の[Ca]i濃度分布画像について、256階調の[Ca]i濃度対応擬似カラーで表示出来る。この画像変化は、5Hzで観察可能である。現在、画像中の30局所の変化を連続記録可能なプログラムを開発し、その有効性を検定している。 (2)本システムやfura-2-front-surface fluorimetry を利用し、次のような研究成果を挙げた。 (1)豚大動脈弁の内皮細胞に光学的方法を応用し、Diadenosine tetraphosphate(AP_4A)は内皮細胞の細胞内Ca放出および細胞外Ca流入によって[Ca]iを上昇させ、NO産生を増加させることを証明した。[Ca]i上昇作用には、P_<2Y>およびP_<2U>受容体が関与していた。同程度の[Ca]i上昇で比較した場合、ATPに比べて、AP_4はより多量のNOを産生した。 (2)内皮を除去した豚冠動脈中膜標本にfront-surface fluorimetry 法およびα毒素処理によるスキン法を応用し、vasoactive intestinal peptide(VIP)の冠動脈弛緩機序について検討した。VIPはK-channelを介した過分極と小胞体へのCa取り込みによる[Ca]i低下、および、G蛋白を介する収縮蛋白のCa感受性低下によって冠動脈を弛緩させることが明らかとなった。 (3)冠動脈にはβ_3-adrenoceptor遺伝子が発現していることを、薬理学的方法およびRT-PCR法を用いて証明した。次に、正常者および虚血性心臓病患者白血球から得られたゲノムDNAとPCR-RFLP法を用いてβ_3-adrenoceptor遺伝子Tr64Arg変異を解析したところ、この変異は正常者に比べて、虚血性心臓病患者では優位に多いことが判った。この遺伝子変異は、冠動脈硬化症の危険因子の一つである可能性が示された。

研究成果

(18件)

すべてその他

すべて文献書誌 (18件)

[文献書誌] H.Jahan: "Endothelin 1 and angiotensin II act as progression but not cpmpetence growth factors in vascular smooth muscle cells." Eur J Pharmacol. 295. 261-269 (1996)
[文献書誌] S.Abe: "Some effects of nipradilol,a β-blocker possessing a nitroxy group,on smooth muscle of the pig coronary artery." Br J Pharmacol. 117. 1707-1715 (1996)
[文献書誌] H.Seguchi: "The mechanism of relaxation induced by atrial natriuretic peptide in the porcine renal artery" Br J Pharmacol. 118. 343-351 (1996)
[文献書誌] Y.Miyagi: "Dual regulation of cerebrovascular tone by UTP:P_<2U> receptormediated contraction and endothelium-dependent relaxation." Br J Pharmacol. 118. 847-856 (1996)
[文献書誌] C.Sakihara: "Expression of calponin mRNA in porcine aortic endothelial cells." Biochem Biophys Res Commun. 222. 195-200 (1996)
[文献書誌] Y.Miyagi: "P_<2U> purinergic activation leads to the cell cycle progression from the G_1 to the S and M phases but not the G_0 to G_1 phasein vascular smooth muscle cells in primary culture." Biochem Biophys Res Commun. 222. 652-658 (1996)
[文献書誌] Y.Miyagi: "P_<2U> receptor is linked to cytosolic Ca^<2+> transient and release of of vasorelaxing factorin bovine endothelial cells in situ." J Physiol. 492.3. 751-761 (1996)
[文献書誌] Y.Fukuizumi: "The effects of calcitonin gene-related peptide on the cytosolic calcium concentration and force in the porcine coronary artery." J Pharmacol Exp Ther. 278. 220-231 (1996)
[文献書誌] Y.Higuchi: "CPA induces a sustained increase in(Ca^<2+>)i of endothelial cells in situ and relaxes porcine coronary artery." Am J Physiol. 270. H2038-H2049 (1996)
[文献書誌] T.Kuga: "Cell cycle-dependent expression of L- and T-type Ca^<2+> currents invascular smooth muscle cells." Circ Res. 79. 14-19 (1996)
[文献書誌] C.Sakihara: "The direct inhibitory effect of chlorpromazineon smooth muscle of the porcine pulmonary artery." Anesthesiology. 85. 616-625 (1996)
[文献書誌] J.Nishimura: "Expression of rhoA and rho kinase mRNAs in porcine vascular smooth muscle." Biochem Biophys Res Commun. 227. 750-754 (1996)
[文献書誌] T.Kai: "Halothane counteracts acetylcholine-induced increase in Ca^<2+> sensitivity of the contractile apparatus in air way smooth muscle." Eur J Pharmacol. 315. 313-318 (1996)
[文献書誌] X.Chem: "Resting load modulates the myofilament Ca^<2+> -sensitivity in rabbit cerebral arteries." J Gereb Blood Flow Metab. (in press).
[文献書誌] G.Hiroishi: "The differential effects of diltiazem and nitroglycerin on cytosolic Ca concentration and on force in the bovine ophthalmic artery." Invest Ophthalmolol Vis Sci. (in press).
[文献書誌] J.Nishimura: "Adrenomrdullin relaxes particular smooth muscles despite a general expression of its mRNA in smooth muscle,endothelial and epithelial cells" Br J Pharmacol. (in press).
[文献書誌] D.Eguchi: "Mechanism of contraction induced by bradykinin in the rabbit saphenous vein." Br J Phaemacol. (in press).
[文献書誌] N.Niiro: "Up-regulation of RHO A and RHO-Kinase mRNA in the rat myometrium during pregnancy." Biochem Biophys Res Commun. (in press).

1996 年度 実績報告書

血管内皮・平滑筋細胞機能を細胞分子レベルで生体連続観察できるシステムの開発

研究代表者

金出 英夫 九州大学, 医学部, 教授 (80038851)

研究成果

[文献書誌] H.Jahan: "Endothelin 1 and angiotensin II act as progression but not cpmpetence growth factors in vascular smooth muscle cells." Eur J Pharmacol. 295. 261-269 (1996)

[文献書誌] S.Abe: "Some effects of nipradilol,a β-blocker possessing a nitroxy group,on smooth muscle of the pig coronary artery." Br J Pharmacol. 117. 1707-1715 (1996)

[文献書誌] H.Seguchi: "The mechanism of relaxation induced by atrial natriuretic peptide in the porcine renal artery" Br J Pharmacol. 118. 343-351 (1996)

[文献書誌] Y.Miyagi: "Dual regulation of cerebrovascular tone by UTP:P_<2U> receptormediated contraction and endothelium-dependent relaxation." Br J Pharmacol. 118. 847-856 (1996)

[文献書誌] C.Sakihara: "Expression of calponin mRNA in porcine aortic endothelial cells." Biochem Biophys Res Commun. 222. 195-200 (1996)

[文献書誌] Y.Miyagi: "P_<2U> purinergic activation leads to the cell cycle progression from the G_1 to the S and M phases but not the G_0 to G_1 phasein vascular smooth muscle cells in primary culture." Biochem Biophys Res Commun. 222. 652-658 (1996)

[文献書誌] Y.Miyagi: "P_<2U> receptor is linked to cytosolic Ca^<2+> transient and release of of vasorelaxing factorin bovine endothelial cells in situ." J Physiol. 492.3. 751-761 (1996)

[文献書誌] Y.Fukuizumi: "The effects of calcitonin gene-related peptide on the cytosolic calcium concentration and force in the porcine coronary artery." J Pharmacol Exp Ther. 278. 220-231 (1996)

[文献書誌] Y.Higuchi: "CPA induces a sustained increase in(Ca^<2+>)i of endothelial cells in situ and relaxes porcine coronary artery." Am J Physiol. 270. H2038-H2049 (1996)

[文献書誌] T.Kuga: "Cell cycle-dependent expression of L- and T-type Ca^<2+> currents invascular smooth muscle cells." Circ Res. 79. 14-19 (1996)

[文献書誌] C.Sakihara: "The direct inhibitory effect of chlorpromazineon smooth muscle of the porcine pulmonary artery." Anesthesiology. 85. 616-625 (1996)

[文献書誌] J.Nishimura: "Expression of rhoA and rho kinase mRNAs in porcine vascular smooth muscle." Biochem Biophys Res Commun. 227. 750-754 (1996)

[文献書誌] T.Kai: "Halothane counteracts acetylcholine-induced increase in Ca^<2+> sensitivity of the contractile apparatus in air way smooth muscle." Eur J Pharmacol. 315. 313-318 (1996)

[文献書誌] X.Chem: "Resting load modulates the myofilament Ca^<2+> -sensitivity in rabbit cerebral arteries." J Gereb Blood Flow Metab. (in press).

[文献書誌] G.Hiroishi: "The differential effects of diltiazem and nitroglycerin on cytosolic Ca concentration and on force in the bovine ophthalmic artery." Invest Ophthalmolol Vis Sci. (in press).

[文献書誌] J.Nishimura: "Adrenomrdullin relaxes particular smooth muscles despite a general expression of its mRNA in smooth muscle,endothelial and epithelial cells" Br J Pharmacol. (in press).

[文献書誌] D.Eguchi: "Mechanism of contraction induced by bradykinin in the rabbit saphenous vein." Br J Phaemacol. (in press).

[文献書誌] N.Niiro: "Up-regulation of RHO A and RHO-Kinase mRNA in the rat myometrium during pregnancy." Biochem Biophys Res Commun. (in press).

1996 年度実績報告書

金出英夫九州大学, 医学部, 教授 (80038851)