Nox4- and Nox2-dependent oxidant production is required for VEGF-induced SERCA cysteine-674 S-glutathiolation and endothelial cell migration.
Nox4- and Nox2-dependent oxidant production is required for VEGF-induced SERCA cysteine-674 S-glutathiolation and endothelial cell migration.
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DOI：
10.1016/j.freeradbiomed.2012.10.546
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发表时间：
2012-12-15
影响因子：
7.4
通讯作者：
Cohen, Richard A.
中科院分区：
文献类型：
--
作者：
Evangelista, Alicia M.;Thompson, Melissa D.;Bolotina, Victoria M.;Tong, XiaoYong;Cohen, Richard A.
内皮细胞（EC）对VEGF的迁移反应是生理性和病理性血管生成的关键步骤。尽管VEGF信号传导已被广泛研究，但VEGF依赖性活性氧（ROS）产生影响EC信号传导的机制尚不清楚。本研究的目的是阐明Nox2-和nox4依赖性ROS在vegf介导的EC Ca2+调节和迁移中的作用。VEGF诱导人主动脉EC在6小时内向抓伤创面迁移，过氧化氢酶或SOD的过表达均可抑制这种迁移。过氧化氢酶能抑制微摩尔浓度H2O2对EC的刺激，但SOD也能抑制。VEGF和H2O2都增加了SERCA2b的s -谷胱甘肽化，增加了Ca2+内流到EC，这些事件可以通过过氧化氢酶过表达或SERCA2b过表达来阻断，其中反应性半胱氨酸-674突变为丝氨酸。在确定vegf介导的ROS产生的来源时，我们的研究表明，特异性敲低Nox2或Nox4均可抑制vegf诱导的SERCA、Ca2+内流和EC迁移的s -谷胱甘肽化。H2O2处理诱导SERCA和EC Ca2+内流的s -谷胱甘肽化，克服了Nox4的敲低，而不是Nox2， Amplex Red测量表明Nox4是H2O2的来源。这些结果表明，VEGF通过增加SERCA的s -谷胱甘肽和Ca2+内流，以Nox4-和h2o2依赖的方式刺激EC迁移，需要下游Nox2。
Endothelial cell (EC) migration in response to VEGF is a critical step in both physiological and pathological angiogenesis. Although VEGF signaling has been extensively studied, the mechanisms by which VEGF-dependent reactive oxygen species (ROS) production affects EC signaling are not well-understood. The aim of this study was to elucidate the involvement of Nox2- and Nox4-dependent ROS in VEGF-mediated EC Ca2+ regulation and migration. VEGF induced migration of human aortic EC into a scratch wound over 6 hours that was inhibited by overexpression of either catalase or SOD. EC stimulation by micromolar concentrations of H2O2 was inhibited by catalase, but also unexpectedly by SOD. Both VEGF and H2O2 increased S-glutathiolation of SERCA2b and increased Ca2+ influx into EC, and these events could be blocked by overexpression of catalase or overexpression of SERCA2b in which the reactive cysteine-674 was mutated to a serine. In determining the source of VEGF-mediated ROS production, our studies show that specific knock down of either Nox2 or Nox4 inhibited VEGF-induced S-glutathiolation of SERCA, Ca2+ influx, and EC migration. Treatment with H2O2 induced S-glutathiolation of SERCA and EC Ca2+ influx, overcoming the knockdown of Nox4, but not Nox2, and Amplex Red measurements indicated that Nox4 is the source of H2O2. These results demonstrate that VEGF stimulates EC migration through increased S-glutathiolation of SERCA and Ca2+ influx in a Nox4- and H2O2-dependent manner, requiring Nox2 downstream.
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影响因子：
7.4
作者：
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通讯作者：
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