HSP90 and Akt modulate Ang-1-induced angiogenesis via NO in coronary artery endothelium
HSP90 and Akt modulate Ang-1-induced angiogenesis via NO in coronary artery endothelium
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DOI：
10.1152/japplphysiol.00728.2003
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发表时间：
2004-02-01
影响因子：
3.3
通讯作者：
Meyrick, B
中科院分区：
文献类型：
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作者：
Chen, JX;Lawrence, ML;Meyrick, B
本研究探讨了热休克蛋白（HSP） 90结合一氧化氮（NO）、内皮NO合成酶（eNOS）和PI3K-Akt调节血管生成素(Ang)-1诱导的猪冠状动脉内皮细胞（PCAEC）血管生成的概念。暴露于Ang-1 （250 ng/ml）长达2小时的时间导致eNOS Ser 1177位点磷酸化的时间依赖性增加，这种增加发生在5分钟，并在60分钟达到峰值。这伴随着一氧化氮释放的逐渐增加。Ang-1还能刺激HSP90与eNOS结合，并显著增加Akt磷酸化。用1马克/毫升格尔达霉素（HSP90的特异性抑制剂）或500纳米wortmannin（一种特异性磷脂酰肌醇3 （PI3）激酶（PI3K）抑制剂）预处理细胞30分钟，可显著减弱ang -1刺激的eNOS磷酸化和NO的产生。暴露于Ang-1导致内皮细胞迁移、管状形成和PCAEC球体发芽增加，而药物阻断HSP90功能或抑制PI3K-Akt通路完全消除了这些作用。n - g-硝基- l -精氨酸甲酯（2.5 mM）对一氧化氮合酶的抑制也显著降低了ang -1诱导的血管生成。我们得出结论，刺激的HSP90结合eNOS和PI3-Akt通路的激活有助于ang -1诱导的eNOS磷酸化、NO生成和PCAEC血管生成。
This study examines the notion that heat shock protein (HSP) 90 binding to nitric oxide ( NO), endothelial NO synthase ( eNOS), and PI3K-Akt regulate angiopoietin (Ang)-1-induced angiogenesis in porcine coronary artery endothelial cells (PCAEC). Exposure to Ang-1 ( 250 ng/ml) for periods up to 2 h resulted in a time-dependent increase in eNOS phosphorylation at Ser 1177 that occurred by 5 min and peaked at 60 min. This was accompanied by a gradual increase in NO release. Ang-1 also led to stimulation of HSP90 binding to eNOS and a significant increase in Akt phosphorylation. Thirty minutes of pretreatment of cells with either 1 mug/ml geldanamycin ( a specific inhibitor of HSP90) or 500 nM wortmannin [ a specific phosphatidylinositol 3 (PI3)-kinase (PI3K) inhibitor] significantly attenuated Ang-1-stimulated eNOS phosphorylation and NO production. Exposure to Ang-1 caused an increase in endothelial cell migration, tube formation, and sprouting from PCAEC spheroids, and pharmacological blockage of HSP90 function or inhibition of PI3K-Akt pathway completely abolished these effects. Inhibition of nitric oxide synthase by N-G-nitro-L-arginine methyl ester (2.5 mM) also resulted in a significant decrease in Ang-1-induced angiogenesis. We conclude that stimulated HSP90 binding to eNOS and activation of the PI3-Akt pathway contribute to Ang-1-induced eNOS phosphorylation, NO production, and angiogenesis in PCAEC.