Vascular endothelial growth factor signals endothelial cell production of nitric oxide and prostacyclin through Flk-1/KDR activation of c-Src
Vascular endothelial growth factor signals endothelial cell production of nitric oxide and prostacyclin through Flk-1/KDR activation of c-Src
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DOI：
10.1074/jbc.274.35.25130
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发表时间：
1999-08-27
影响因子：
4.8
通讯作者：
Caldwell, RB
中科院分区：
文献类型：
--
作者：
He, H;Venema, VJ;Caldwell, RB
血管内皮生长因子(VEGF)是一种强大的内皮细胞特异性有丝分裂原，通过涉及一氧化氮(NO)和前列环素(PGI(2))的自分泌机制促进血管生成、血管高通透性和血管扩张。本实验采用免疫沉淀和免疫分析方法研究了血管内皮生长因子诱导培养的内皮细胞产生NO和PGI(2)的信号通路。结果表明，血管内皮细胞生长因子刺激Flk-1/KDR受体与c-Src形成复合体，而src激活是血管内皮生长因子诱导磷脂酶C-1活化和1，4，5-三磷酸肌醇形成所必需的。报告细胞分析显示，血管内皮细胞生长因子促进NO生成的作用类似于50倍，在5-20分钟达到峰值。这种作用是由c-Src的Flk-1/KDR激活启动的信号级联反应介导的，导致磷脂酶C-Gamma 1的激活、1，4，5-三磷酸肌醇的形成、[Ca~(2+)](I)的释放和一氧化氮合酶的激活。免疫分析显示，血管内皮生长因子诱导的6-酮-前列腺素F-1α的形成是PGI(2)产生的指标，增加了3-4倍，在45-60分钟达到峰值。PGI(2)信号通路通过释放[Ca~(2+)](I)跟随NO通路，但在NOS激活之前发生分化，还需要丝裂原激活的蛋白激酶的激活。这些结果表明，NO和PGI(2)在介导血管内皮生长因子的作用中平行发挥作用。
Vascular endothelial growth factor (VEGF) is a potent endothelial cell-specific mitogen that promotes angiogenesis, vascular hyperpermeability, and vasodilation by autocrine mechanisms involving nitric oxide (NO) and prostacyclin (PGI(2)) production, These experiments used immunoprecipitation and immunoassay procedures to characterize the signaling pathways by which VEGF induces NO and PGI(2) formation in cultured endothelial cells. The data showed that VEGF stimulates complex formation of the flk-1/kinase-insert domain-containing receptor (KDR) VEGF receptor with c-Src and that Src activation is required for VEGF induction of phospholipase C gamma 1 activation and inositol 1,4,5-trisphosphate formation. Reporter cell assays showed that VEGF promotes a similar to 50-fold increase in NO formation, which peaks at 5-20 min. This effect is mediated by a signaling cascade initiated by flk-1/KDR activation of c-Src, leading to phospholipase C gamma 1 activation, inositol 1,4,5-trisphosphate formation, release of [Ca2+](i) and nitric oxide synthase activation. Immunoassays of VEGF-induced 6-keto prostaglandin F-1 alpha formation as an indicator of PGI(2) production revealed a 3-4-fold increase that peaked at 45-60 min. The PGI(2) signaling pathway follows the NO pathway through release of [Ca2+](i), but diverges prior to NOS activation and also requires activation of mitogen-activated protein kinase, These results suggest that NO and PGI(2) function in parallel in mediating the effects of VEGF.