PKA and Epac activation mediates cAMP-induced vasorelaxation by increasing endothelial NO production

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• 作者：Veró ; nica Garcí ; a-Morales ; Andrea Cuí ; ñ ; as ; Jacobo Elí ; es¹ ; Manuel Campos-Toimil ; ^{manuel.campos@usc.es" class="auth_mail}
• 关键词：[Ca2  ; +]c ; cytoplasmic Ca2  ; + concentration ; cAMP ; 3&prime ; 5&prime ; -cyclic adenosine monophosphate ; DAF-2 ; 4 ; 5-diaminofluorescein ; DAF-2-DA ; 4 ; 5-diaminofluorescein diacetate ; db-cAMP ; dibutyryl cyclic AMP ; DMSO ; dimethyl sulfoxide ; eNOS ; endothelial NO synthase ; Epac ; exchange protein directly activated by cAMP ; FBS ; fetal bovine serum ; HUVEC ; human umbilical vein endothelial cells ; KBS ; Krebs bicarbonate solution ; L-NAME ; N蠅-nitro-l-arginine methyl ester ; PKA ; cyclic-AMP protein kinas
• 刊名：Vascular Pharmacology
• 出版年：March, 2014
• 年：2014
• 卷：60
• 期：3
• 页码：95-101
• 全文大小：625 K

文摘

Vascular relaxation induced by 3鈥?5鈥?cyclic adenosine monophosphate (cAMP) is both endothelium-dependent and endothelium-independent, although the underlying signaling pathways are not fully understood. Aiming to uncover potential mechanisms, we performed contraction-relaxation experiments on endothelium-denuded and intact rat aorta rings and measured NO levels in isolated human endothelial cells using single cell fluorescence imaging. The vasorelaxant effect of forskolin, an adenylyl cyclase activator, was decreased after selective inhibitor of protein kinase A (PKA), a cAMP-activated kinase, or L-NAME, an endothelial nitric oxide synthase (eNOS) inhibitor, only in intact aortic rings. Both selective activation of PKA with 6-Bnz-cAMP and exchange protein directly activated by cAMP (Epac) with 8-pCPT-2鈥测€?em>O-Me-cAMP significantly relaxed phenylephrine-induced contractions. The vasorelaxant effect of the Epac activator, but not that of the PKA activator, was reduced by endothelium removal. Forskolin, dibutyryl cAMP (a cAMP analogue), 6-Bnz-cAMP and 8-pCPT-2鈥测€?em>O-Me-cAMP increased NO levels in endothelial cells and the forskolin effect was significantly inhibited by inactivation of both Epac and PKA, and eNOS inhibition. Our results indicate that the endothelium-dependent component of forskolin/cAMP-induced vasorelaxation is partially mediated by an increase in endothelial NO release due to an enhanced eNOS activity through PKA and Epac activation in endothelial cells.