Role of c-Src in the regulation of vascular contraction and Ca2+ signaling by angiotensin II in human vascular smooth muscle cells

被引:86
作者
Touyz, RM
Wu, XH
He, G
Park, JB
Chen, X
Vacher, J
Rajapurohitam, V
Schiffrin, EL
机构
[1] Univ Montreal, Clin Res Inst Montreal, Lab Cellular Interact & Dev, Montreal, PQ H2W 1R7, Canada
[2] Sungkyunkwan Univ, Sch Med, Samsung Cheil Hosp, Seoul, South Korea
[3] Sungkyunkwan Univ, MRC, Multidisciplinary Res Grp Hypertens, Seoul, South Korea
关键词
signal transduction; calcium; contractility;
D O I
10.1097/00004872-200103000-00012
中图分类号
R6 [外科学];
学科分类号
1002 ; 100210 ;
摘要
Objective Tyrosine kinases, typically associated with growth-signaling pathways, also play a role in Ang II-stimulated vascular contraction. However the specific kinases involved are unclear. We hypothesize here that c- Src, a non-receptor tyrosine kinase, is an important upstream regulator of vascular smooth muscle cell (VSMC) Ca2+ signaling and associated vascular contraction induced by Ang II, Methods Cultured VSMCs from resistance arteries of healthy subjects were studied. Human VSMCs electroporated with anti-c-Src antibody and c-Src-deficient VSMCs from small arteries of c-Src knockout mice (Src-/- mVSMCs) were also investigated. Intracellular free Ca2+ concentration ([Ca2+](i)), c-Src activity and IP3 production were measured by fura 2, immunoblot and radioimmunoassay respectively. Contraction was examined in intact rat small arteries. Results Ang II rapidly increased VSMC c-Src activity, with peak responses obtained at 1 min. Ang II induced a biphasic [Ca2+](i) response (E-max = 636 +/- 123 nmol/l). The initial [Ca2+](i) transient, mediated primarily by Ca2+ mobilization, was dose-dependently attenuated by the selective Src inhibitor, PP2, but not by PP3 (inactive analogue). Ang II-elicited [Ca2+](i) responses were blunted in cells electroporated with anti-c-Src antibodies and in c Src-/-mVSMCs, Src inhibition decreased Ang II-induced generation of IP3 in human VSMCs. Ang II dose-dependently increased vascular contraction (E-max = 40 +/- 6.5%). These responses were attenuated by PP2 (E-max = 7.8 +/- 0.08%) but not by PP3 (E-max = 35 +/- 4.5%). Conclusions Our findings identify c-Src as an important regulator of VSMC [Ca2+](i) signaling and implicate a novel contractile role for this non-receptor tyrosine kinase in Ang Ii-stimulated vascular smooth muscle. I Hypertens 19:441-449 (C) 2001 Lippincott Williams & Wilkins.
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页码:441 / 449
页数:9
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