VEGF诱导的血管生成和血管通透性信号分化:抑制蛋白激酶C抑制VEGF诱导的血管生成,但促进VEGF诱导的no依赖性血管通透性

I. Spyridopoulos, Corinne Luedemann, Donghui Chen, M. Kearney, Dongfen Chen, T. Murohara, N. Principe, J. Isner, Douglas Losordo
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引用次数: 69

摘要

血管内皮生长因子(VEGF)通过多种机制促进血管生成,包括刺激内皮细胞增殖和迁移以及增加血管通透性。尽管其有丝分裂活性主要由蛋白激酶C (PKC)的2-异构体介导,但对其转导其他生理特性的信号通路知之甚少。因此,我们使用一种新的抑制剂分子来检测PKC异构体的作用&agr;bgr;介导vegf诱导的血管生成和血管通透性。由于PKC的常规抑制剂,如staurosporine或calphostin C,也抑制多种其他蛋白激酶,我们使用了一种新的化合物来特异性抑制PKC。一种肉豆蔻酰化肽,它模仿PKC-&agr的假底物基序;和-&bgr;已被证明是PKC的高选择性和细胞渗透性抑制剂。正如预期的那样,阻断导致体外vegf诱导的内皮细胞增殖消失。在体内,vegf诱导的血管生成被肉豆蔻酰基化肽破坏。令人惊讶的是,PKC的选择性抑制通过no依赖机制诱导了体内血管通透性。此外,PKC抑制导致内皮细胞NO合成酶(NOS)活性升高6.4倍。我们的研究结果表明,激活PKC是vegf诱导的增殖和血管生成所需的主要信号通路,而通过阻断PKC可以增强血管通透性。钙依赖性PKC本身的抑制导致NOS的诱导。尽管NOS是vegf诱导血管生成的下游靶点,但PKC抑制对其的诱导不足以促进新生血管的形成。这些结果表明,VEGF诱导的血管生成和血管通透性是由最终分化的机制介导的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Divergence of Angiogenic and Vascular Permeability Signaling by VEGF: Inhibition of Protein Kinase C Suppresses VEGF-Induced Angiogenesis, but Promotes VEGF-Induced, NO-Dependent Vascular Permeability
Vascular endothelial growth factor (VEGF) promotes angiogenesis by a variety of mechanisms including stimulation of endothelial cell proliferation and migration and increasing vascular permeability. Although its mitogenic activity is mediated primarily by the &bgr;2-isoforms of protein kinase C (PKC), little is known about the signaling pathways transducing its other physiological properties. Accordingly, we used a novel inhibitor molecule to examine the role of PKC isoforms &agr; and &bgr; in mediating VEGF-induced angiogenesis and vascular permeability. Because conventional inhibitors of PKC, such as staurosporine or calphostin C, also inhibit a variety of other protein kinases, we used a novel compound to specifically inhibit PKC. A myristoylated peptide, which mimics the pseudosubstrate motif of PKC-&agr; and -&bgr; subtypes, has been shown to be a highly selective and cell-permeable inhibitor of PKC. Blocking led, as expected, to abrogation of VEGF-induced endothelial cell proliferation in vitro. In vivo, VEGF-induced angiogenesis was impaired by myristoylated peptide. Surprisingly, selective inhibition of PKC induced vascular permeability in vivo via a NO-dependent mechanism. Moreover, PKC inhibition led to a 6.4-fold induction of NO synthase (NOS) activity in endothelial cells. Our findings demonstrate that activation of PKC is a major signaling pathway required for VEGF-induced proliferation and angiogenesis, whereas vascular permeability was enhanced by blocking PKC. Inhibition of calcium-dependent PKC by itself led to induction of NOS. Although NOS is a downstream target for VEGF-induced angiogenesis, its induction by PKC inhibition was not sufficient to promote neovascularization. These results reveal that angiogenesis and vascular permeability induced by VEGF are mediated by mechanisms which ultimately diverge.
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