Novel role of gp91phox-containing NAD(P)H oxidase in vascular endothelial growth factor-induced signaling and angiogenesis (Retracted Article)
Novel role of gp91phox-containing NAD(P)H oxidase in vascular endothelial growth factor-induced signaling and angiogenesis (Retracted Article)
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DOI：
10.1161/01.res.0000046227.65158.f8
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发表时间：
2002-12-13
影响因子：
20.1
通讯作者：
Alexander, RW
中科院分区：
文献类型：
--
作者：
Ushio-Fukai, M;Tang, Y;Alexander, RW
血管内皮生长因子(VEGF)通过刺激内皮细胞的增殖和迁移，主要通过受体酪氨酸激酶VEGF受体2(Flk1KDR)诱导血管生成。由NAD(P)H氧化酶产生的活性氧物种(ROS)在血管细胞调节的许多方面都是至关重要的，而小GTP酶rac1和gp91(Phox)x都是内皮NAD(P)H氧化酶复合体的关键成分。然而，NAD(P)H氧化酶在血管内皮生长因子诱导的血管生成中的作用尚不清楚。在本研究中，我们利用电子自旋共振波谱证实，在人脐静脉内皮细胞中，血管内皮生长因子可刺激O-2(.-)的产生，这种作用可被NAD(P)H氧化酶抑制剂二苯碘、显性负性rac1(N17rac1)基因的过表达和gp91(Phox)反义寡核苷酸的过表达所抑制。抗氧化剂，包括N-乙酰半胱氨酸(NAC)、各种NAD(P)H氧化酶抑制剂和N17rac1，不仅显著抑制血管内皮生长因子诱导的KDR酪氨酸磷酸化，而且显著抑制内皮细胞的增殖和迁移。重要的是，在转染gp91(Phox)反义寡核苷酸的细胞中，血管内皮生长因子的这些作用被显著抑制。相反，ROS不参与介导1-磷酸鞘氨醇(SIP)对内皮细胞的这些影响。海绵植入实验表明，NAC处理的野生型小鼠和gp91(Phox-/-)小鼠的血管生成显著减少，而不是S1P-诱导的血管生成显著减少，这表明含有gp91(Phox)的NAD(P)H氧化酶产生的ROS在体内的血管生成中起重要作用。这些研究表明，血管内皮生长因子诱导的内皮细胞信号转导和血管生成在血管生成受体水平上受到还原/氧化环境的严格控制，为NAD(P)H氧化酶作为血管生成依赖型疾病的潜在治疗靶点提供了新的见解。
Vascular endothelial growth factor (VEGF) induces angiogenesis by stimulating endothelial cell proliferation and migration, primarily through the receptor tyrosine kinase VEGF receptor2 (Flk1KDR). Reactive oxygen species (ROS) derived from NAD(P)H oxidase are critically important in many aspects of vascular cell regulation, and both the small GTPase Rac1 and gp91(phox) x are critical components of the endothelial NAD(P)H oxidase complex. A role of NAD(P)H oxidase in VEGF-induced angiogenesis, however, has not been defined. In the present study, electron spin resonance spectroscopy is utilized to demonstrate that VEGF stimulates O-2(.-) production, which is inhibited by the NAD(P)H oxidase inhibitor, diphenylene iodonium, as well as by overexpression of dominant-negative Rac1 (N17Rac1) and transfection of gp91(phox) antisense oligonucleotides in human umbilical vein endothelial cells (ECs). Antioxidants, including N-acetylcysteine (NAC), various NAD(P)H oxidase inhibitors, and N17Rac1 significantly attenuate not only VEGF-induced KDR tyrosine phosphorylation but also proliferation and migration of ECs. Importantly, these effects of VEGF are dramatically inhibited in cells transfected with gp91(phox) antisense oligonucleotides. By contrast, ROS are not involved in mediating these effects of sphingosine 1-phosphate (SIP) on ECs. Sponge implant assays demonstrate that VEGF-, but not S1P-, induced angiogenesis is significantly reduced in wild-type mice treated with NAC and in gp91(phox-/-) mice, suggesting that ROS derived from gp91(phox)-containing NAD(P)H oxidase play an important role in angiogenesis in vivo. These studies indicate that VEGF-induced endothelial cell signaling and angiogenesis is tightly controlled by the reduction/oxidation environment at the level of VEGF receptor and provide novel insights into the NAD(P)H oxidase as a potential therapeutic target for angiogenesis-dependent diseases.