Vascular endothelial growth factor governs endothelial nitric-oxide synthase expression via a KDR/Flk-1 receptor and a protein kinase C signaling pathway
Vascular endothelial growth factor governs endothelial nitric-oxide synthase expression via a KDR/Flk-1 receptor and a protein kinase C signaling pathway
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DOI：
10.1074/jbc.274.46.33057
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发表时间：
1999-11-12
影响因子：
4.8
通讯作者：
Zioncheck, TF
中科院分区：
文献类型：
--
作者：
Shen, BQ;Lee, DY;Zioncheck, TF
血管内皮生长因子（VEGF）调控内皮型一氧化氮合酶（eNOS）表达的机制目前尚不清楚。在此，我们报道了VEGF处理牛肾上腺皮质内皮细胞导致eNOS蛋白和活性增加B倍，内皮NOS表达在恒定VEGF暴露（500 pM）2天后达到最大，并在第5天下降到基线水平。如果VEGF与L-N-G-硝基精氨酸甲酯共孵育，竞争性eNOS抑制剂。加入一氧化氮供体S-亚硝基-N-乙酰青霉胺（S-nitroso-N-acetylpenicillamine）可阻止VEGF诱导的eNOS表达上调。这些数据表明，一氧化氮参与了调节eNOS表达的负反馈机制。各种方法被用来研究两种高亲和力VEGF受体在eNOS上调中的作用。HDR受体选择性突变体增加eNOS表达，而Flt-1受体选择性突变体没有。此外，VEGF治疗增加eNOS表达在KDR，但不是在Flt-1受体转染的猪主动脉内皮细胞系。SU 1498，一种选择性的KDR受体酪氨酸激酶抑制剂，阻断eNOS的上调，从而提供了进一步的证据，KDR受体信号eNOS的上调，最后，治疗肾上腺皮质内皮细胞与VEGF或佛波酯导致蛋白激酶C的激活和eNOS的表达升高，而蛋白激酶C的抑制与亚型特异性抑制剂废除VEGF诱导的eNOS上调。总之，这些数据表明，VEGF通过激活KDR受体酪氨酸激酶和下游蛋白激酶C信号通路增加eNOS表达。
The mechanism by which vascular endothelial growth factor (VEGF) regulates endothelial nitric-oxide synthase (eNOS) expression is presently unclear. Here we report that VEGF treatment of bovine adrenal cortex endothelial cells resulted in a B-fold increase in both eNOS protein and activity, Endothelial NOS expression was maximal following 2 days of constant VEGF exposure (500 pM) and declined to base-line levels by day 5, The elevated eNOS protein level was sustained over the time course if VEGF was co-incubated with L-N-G-nitroarginine methyl ester, a competitive eNOS inhibitor. Addition of S-nitroso-N-acetylpenicillamine, a nitric oxide donor, prevented VEGF-induced eNOS up-regulation, These data suggest that nitric oxide participates in a negative feedback mechanism regulating eNOS expression. Various approaches were used to investigate the role of the two high affinity VEGF receptors in eNOS up-regulation. A HDR receptor-selective mutant increased eNOS expression, whereas an Flt-1 receptor-selective mutant did not. Furthermore, VEGF treatment increased eNOS expression in a KDR but not in an Flt-1 receptor-transfected porcine aorta endothelial cell line. SU1498, a selective inhibitor of the KDR receptor tyrosine kinase, blocked eNOS up-regulation thus providing further evidence that the KDR receptor signals for eNOS up-regulation, Finally, treatment of adrenal cortex endothelial cells with VEGF or phorbol ester resulted in protein kinase C activation and elevated eNOS expression, whereas inhibition of protein kinase C with isoform-specific inhibitors abolished VFGF-induced eNOS up-regulation. Taken together, these data demonstrate that VEGF increases eNOS expression via activation of the KDR receptor tyrosine kinase and a downstream protein kinase C signaling pathway.