Angiotensin II upregulates Ca(V)1.2 protein expression in cultured arteries via endothelial H(2)O(2) production.
Abstract
We previously reported that angiotensin II caused an endothelial-dependent increase in L-type voltage-dependent Ca(2+) channel (Ca(V)1.2) in cultured arteries, but the signaling pathways are not clear. Endothelial damage was generated by brief intra-arterial perfusion with 0.3% CHAPS. Ca(V)1.2 expression, function and H(2)O(2) were measured by Western blot, tension recording and Amplex Red H(2)O(2) assay kit, respectively. Angiotensin II dose-dependently upregulated Ca(V)1.2 expression in endothelium-intact arteries. The angiotensin II upregulation of Ca(V)1.2 expression in endothelium-intact arteries was blocked by NAD(P)H oxidase inhibitor diphenyleneiodonium (DPI), apocynin, a more specific NAD(P)H oxidase inhibitor gp91ds-tat and also by catalase. H(2)O(2) similarly upregulated Ca(V)1.2 expression in endothelium-intact and endothelium-damaged arteries, and the latter effect was also blocked by DPI and apocynin. Angiotensin II increased H(2)O(2) production by endothelium-intact but not by endothelium-damaged arteries, and this effect was blocked by apocynin, catalase and gp91ds-tat. The upregulation of Ca(V)1.2 by angiotensin II and H(2)O(2) is accompanied by an increased tension response to KCl and the Ca(2+) channel activa...Continue Reading
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