1. eNOS activation induced by a polyphenol-rich grape skin extract in porcine coronary arteries.
- Author
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Madeira SV, Auger C, Anselm E, Chataigneau M, Chataigneau T, Soares de Moura R, and Schini-Kerth VB
- Subjects
- Animals, Cell Survival drug effects, Cells, Cultured, Coronary Vessels cytology, Coronary Vessels enzymology, Endothelial Cells cytology, Endothelial Cells enzymology, Enzyme Activation drug effects, MAP Kinase Signaling System drug effects, MAP Kinase Signaling System physiology, Nitric Oxide metabolism, Organ Culture Techniques, Oxidation-Reduction, Phosphorylation drug effects, Phosphorylation physiology, Polyphenols, Proto-Oncogene Proteins c-akt metabolism, Reactive Oxygen Species metabolism, Swine, Vasodilation drug effects, Vasodilation physiology, src-Family Kinases metabolism, Coronary Vessels drug effects, Endothelial Cells drug effects, Flavonoids pharmacology, Nitric Oxide Synthase Type III metabolism, Phenols pharmacology, Plant Extracts pharmacology, Vitis
- Abstract
Background/aims: Drinking red wine is associated with a decreased mortality from coronary heart diseases. This study examined whether polyphenols contained in a grape skin extract (GSE) triggered the endothelial formation of nitric oxide (NO) and investigated the underlying mechanism., Methods: Vascular reactivity was assessed in organ chambers using porcine coronary artery rings in the presence of indomethacin (a cyclooxygenase inhibitor) and charybdotoxin plus apamin (inhibitors of endothelium-derived hyperpolarizing factor-mediated responses). The phosphorylation level of Src, Akt and endothelial NO synthase (eNOS) were assessed by Western blot analysis, and the formation of reactive oxygen species (ROS) was investigated using dihydroethidine and dichlorodihydrofluorescein., Results: GSE-induced endothelium-dependent relaxations were abolished by N(G)-nitro-L-arginine (an eNOS inhibitor) and ODQ (a soluble guanylyl cyclase inhibitor), and they were reduced by MnTMPyP, polyethyleneglycol catalase, PP2 (an inhibitor of Src kinase) and wortmannin (an inhibitor of phosphoinositide 3-kinase). GSE caused phosphorylation of Src, which was prevented by MnTMPyP. It also caused phosphorylation of Akt and eNOS, which were prevented by MnTMPyP, polyethyleneglycol catalase, PP2, wortmannin and LY294002. GSE elicited the formation of ROS in native and cultured endothelial cells, which was prevented by MnTMPyP., Conclusions: GSE causes endothelium-dependent NO-mediated relaxations of coronary arteries. This effect involves the intracellular formation of ROS in endothelial cells leading to the Src kinase/phosphoinositide 3-kinase/Akt-dependent phosphorylation of eNOS.
- Published
- 2009
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