EDF-1 contributes to the regulation of nitric oxide release in VEGF-treated human endothelial cells
EDF-1 contributes to the regulation of nitric oxide release in VEGF-treated human endothelial cells
复制标题
DOI：
10.1016/j.ejcb.2010.05.001
复制
发表时间：
2010-09-01
影响因子：
6.6
通讯作者：
Maier, Jeanette A. M.
中科院分区：
文献类型：
--
作者：
Leidi, Marzia;Mariotti, Massimo;Maier, Jeanette A. M.
血管内皮生长因子（VEGF）通过触发多种细胞内信号诱导一氧化氮（NO）释放，其中包括钙/钙调素通路和Akt的激活，这些事件诱导内皮NO合成酶（eNOS）活性。由于内皮分化相关因子(EDF)-1是一种钙调素结合蛋白，并在调节内皮功能中发挥作用，我们评估了EDF-1是否与vegf处理的人内皮细胞中eNOS活性的调节有关。虽然VEGF不调节EDF-1的总量，但它促进钙调素与EDF-1的解离，这与eNOS结合的钙调素增加和诱导NO释放有关。为了更好地表征EDF-1在调节vegf诱导的NO释放中的作用，我们在内皮细胞中稳定地沉默了EDF-1。我们在这里表明，沉默EDF-1的内皮细胞比对照组产生更多的NO，并且在响应VEGF时不增加NO的释放。对VEGF不敏感的原因是，即使Akt被激活，细胞也不能使EDF-1磷酸化eNOS Ser（1177）。有趣的是，在沉默EDF-1的细胞中，丝氨酸/苏氨酸磷酸酶PP2A的药理学抑制剂冈田酸可以优先去磷酸化eNOS Ser(1177)，恢复NO释放和eNOS Ser（1177）磷酸化。我们的研究结果表明，EDF-1是人类内皮细胞中eNOS活性复杂调控的新贡献者。(C) 2010 Elsevier GmbH版权所有。
Vascular endothelial growth factor (VEGF) induces nitric oxide (NO) release by triggering multiple intracellular signals, among others the calcium/calmodulin pathway and the activation of Akt, events which induce endothelial NO synthase (eNOS) activity. Because Endothelial Differentiation-related Factor (EDF)-1 is a calmodulin binding protein and plays a role in modulating endothelial functions, we evaluated whether EDF-1 is implicated in the regulation of eNOS activity in VEGF-treated human endothelial cells. While VEGF does not modulate the total amounts of EDF-1, it promotes the dissociation of calmodulin from EDF-1 which correlates with the increase of calmodulin bound to eNOS and the induction of NO release. To better characterize the contribution of EDF-1 to the regulation of VEGF-induced NO release, we stably silenced EDF-1 in endothelial cells. We here show that endothelial cells silencing EDF-1 produce more NO than controls and do not increase NO release in response to VEGF. The insensitivity to VEGF results from the incapability of cells silencing EDF-1 to phosphorylate eNOS Ser(1177), even though Akt is activated. Interestingly, okadaic acid, a pharmacologic inhibitor of the serine/threonine phosphatase PP2A, which preferentially dephosphorylates eNOS Ser(1177), restores NO release and eNOS Ser(1177) phosphorylation in cells silencing EDF-1. Our results suggest EDF-1 as a novel contributor to the complex regulation of eNOS activity in human endothelial cells. (C) 2010 Elsevier GmbH. All rights reserved.