Grape juice causes endothelium-dependent relaxation via a redox-sensitive Src- and Akt-dependent activation of eNOS
Grape juice causes endothelium-dependent relaxation via a redox-sensitive Src- and Akt-dependent activation of eNOS
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DOI：
10.1016/j.cardiores.2006.08.004
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发表时间：
2007-01-15
影响因子：
10.8
通讯作者：
Schini-Kerth, Valerie B.
中科院分区：
文献类型：
--
作者：
Anselm, Eric;Chataigneau, Marta;Schini-Kerth, Valerie B.
目的：一氧化氮（NO）和内皮源性超极化因子（EDHF）的内皮形成增强，被认为有助于适量饮用红酒对冠状动脉疾病的保护作用。本研究描述了康科德葡萄汁（CGJ）对冠状动脉的内皮依赖性松弛作用，CGJ是一种富含葡萄多酚的非酒精来源。方法：将猪冠状动脉环悬挂在器官室中，以测量吲哚美辛存在下等长张力的变化。通过电子自旋共振光谱法评估 NO 形成，并通过蛋白质印迹分析评估培养的内皮细胞中 Src、Akt 和内皮 NO 合酶 (eNOS) 的磷酸化。结果：L-NA 轻微但显着地减少了内皮依赖性 CGJ 松弛，不受 Charybdotoxin (CTX) 加 apamin（APA，EDHF 介导反应的两种抑制剂）的影响，而 L-NA、CTX 加 APA 的组合将最大松弛度降低至约 50%。在 CTX 加 APA 存在的情况下，超氧化物歧化酶 (SOD) 的膜渗透模拟物、MnTMPyP、过氧化氢酶聚乙二醇过氧化氢酶 (PFG-过氧化氢酶) 的膜渗透类似物、PP2（Src 激酶抑制剂）和渥曼青霉素（PI3 激酶抑制剂）显着减少了 CGJ 的松弛。 CGJ 刺激内皮细胞中活性氧的形成以及 N-w-硝基-L-精氨酸、PP2 和渥曼青霉素敏感的 NO 形成。 NO 的形成与 Src、Akt 和 eNOS 的氧化还原敏感且时间依赖性磷酸化有关。结论：CGJ 诱导冠状动脉内皮依赖性松弛，其中涉及 NO 介导的成分，并且在较小程度上还涉及 EDHF 介导的成分。此外，CGJ 诱导的 NO 形成是由于 Src 激酶的氧化还原敏感激活以及随后的 eNOS PI3 激酶/Akt 依赖性磷酸化所致。 (c) 2006 年欧洲心脏病学会。由 Elsevier B.V. 出版。保留所有权利。
Objectives: An enhanced endothelial formation of nitric oxide (NO) and endothelium-derived hyperpolarizing factor (EDHF), is thought to contribute to the protective effect of moderate consumption of red wine on coronary diseases. The present study has characterized endothelium-dependent relaxations to Concord grape juice (CGJ), a non-alcoholic rich source of grape-derived polyphenols, in the coronary artery.Methods: Porcine coronary artery rings were suspended in organ chambers for the measurement of changes in isometric tension in the presence of indomethacin. NO formation was assessed by electron spin resonance spectroscopy, and the phosphorylation of Src, Akt and endothelial NO synthase (eNOS) by Western blot analysis in cultured endothelial cells.Results: Endothelium-dependent relaxations to CGJ were slightly but significantly reduced by L-NA, not affected by charybdotoxin (CTX) plus apamin (APA, two inhibitors of EDHF-mediated responses) whereas the combination of L-NA, CTX plus APA reduced maximal relaxation to about 50%. In the presence of CTX plus APA, relaxations to CGJ were markedly reduced by the membrane permeant mimetic of superoxide dismutase (SOD), MnTMPyP, the membrane permeant analogue of catalase polyethyleneglycol-catalase (PFG-catalase), PP2, an inhibitor of Src kinase, and by wortmannin, an inhibitor of the PI3-kinase. CGJ stimulated the formation of reactive oxygen species and the N-w-nitro-L-arginine-, PP2- and wortmannin-sensitive formation of NO in endothelial cells. The formation of NO was associated with a redox-sensitive and time-dependent phosphorylation of Src, Akt and eNOS.Conclusions: CGJ induces endothelium-dependent relaxations of coronary arteries, which involve a NO-mediated component and also, to a minor extent, an EDHF-mediated component. In addition, CGJ-induced NO formation is due to the redox-sensitive activation of Src kinase with the subsequent PI3-kinase/Akt-dependent phosphorylation of eNOS. (c) 2006 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.