Role of redox signaling and poly (adenosine diphosphate-ribose) polymerase activation in vascular smooth muscle cell growth inhibition by nitric oxide and peroxynitrite.
Role of redox signaling and poly (adenosine diphosphate-ribose) polymerase activation in vascular smooth muscle cell growth inhibition by nitric oxide and peroxynitrite.
复制标题
氧化还原信号传导和聚（腺苷二磷酸核糖）聚合酶激活在一氧化氮和过氧亚硝酸盐抑制血管平滑肌细胞生长中的作用。
DOI：
10.1016/j.jvs.2007.11.006
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发表时间：
2008
影响因子：
4.3
通讯作者：
Sarkar,Rajabrata
中科院分区：
文献类型：
--
作者：
Huang,James;Lin,StephanieC;Nadershahi,Afshin;Watts,StephanieW;Sarkar,Rajabrata
目的作为血管介质，一氧化氮调节血管平滑肌细胞增殖，并可与超氧化物反应形成过氧亚硝酸盐，一种高活性自由基。一氧化氮和过氧亚硝酸盐抑制平滑肌细胞生长的细胞内机制仍不清楚，过氧亚硝酸盐形成在一氧化氮的抗增殖作用中的潜在作用也不清楚。我们试图明确一氧化氮和过氧亚硝酸盐在平滑肌细胞中的细胞内效应和信号传导机制。方法用外源性一氧化氮或过氧亚硝酸盐以及一氧化氮和氧化还原信号通路抑制剂处理培养的大鼠主动脉平滑肌细胞。检测了细胞生长、DNA 合成、细胞凋亡、环鸟苷 3'-5' 单磷酸 (cGMP) 水平、聚二磷酸腺苷 [ADP]-核糖）聚合酶 (PARP) 活性和细胞毒性。通过硝基酪氨酸免疫印迹法测定过氧亚硝酸盐的形成。用一氧化氮/过氧亚硝酸盐和氧化还原剂处理后，评估离体大鼠主动脉环的血管反应性。结果外源性一氧化氮和过氧亚硝酸盐均减少培养的大鼠主动脉平滑肌细胞的生长和DNA合成，但过氧亚硝酸盐诱导的生长停滞是不可逆的，并与细胞凋亡和细胞毒性相关。鸟苷酸环化酶、PARP 活性、丝裂原激活蛋白激酶的抑制或鸟氨酸脱羧酶的旁路不能逆转一氧化氮引起的生长停滞。抗氧化剂 N-乙酰半胱氨酸、抗坏血酸和谷胱甘肽选择性地逆转一氧化氮造成的生长抑制，但不能逆转过氧亚硝酸盐造成的生长抑制。抗氧化剂不会损害平滑肌细胞中一氧化氮诱导的 cGMP 生成或一氧化氮诱导的离体主动脉环的血管舒张。一氧化氮处理不会导致过氧亚硝酸盐的形成，并且超氧化物水平的增加不会引起类似过氧亚硝酸盐的作用。过氧亚硝酸盐诱导的细胞毒性和细胞凋亡不能通过抗氧化剂或 PARP 抑制来逆转，因为过氧亚硝酸盐激活 J774 巨噬细胞中的 PARP，但无法激活平滑肌细胞中的 PARP。 结论 外源性一氧化氮通过独立于过氧亚硝酸盐形成且不同于一氧化氮血管舒张机制的氧化还原敏感机制诱导平滑肌细胞中的可逆细胞抑制。过氧亚硝酸盐不会选择性激活平滑肌细胞中的 PARP，并诱导不依赖氧化还原的平滑肌细胞细胞毒性和凋亡。因此，一氧化氮和过氧亚硝酸盐对平滑肌细胞的抗增殖作用利用具有不同氧化还原敏感性的不同细胞内途径。这些发现与血管疾病的发病机制以及基于一氧化氮的血管疾病治疗的潜在应用相关。
PURPOSEThe vascular mediator, nitric oxide regulates vascular smooth muscle cell proliferation and can react with superoxide to form peroxynitrite, a highly reactive free radical. The intracellular mechanisms by which nitric oxide and peroxynitrite inhibit smooth muscle cell growth remain undefined, as is the potential role of peroxynitrite formation in the antiproliferative effects of nitric oxide. We sought to define the intracellular effects and signaling mechanisms of nitric oxide and peroxynitrite in smooth muscle cells.METHODSCultured rat aortic smooth muscle cells were treated with exogenous nitric oxide or peroxynitrite and inhibitors of nitric oxide and redox signaling pathways. Cell growth, DNA synthesis, apoptosis, cyclic guanosine 3′-5′ monophosphate (cGMP) levels, poly(adenosine diphosphate [ADP]-ribose) polymerase (PARP) activity, and cytotoxicity were assayed. Peroxynitrite formation was determined by nitrotyrosine immunoblotting. Vasoreactivity was assessed in isolated rat aortic rings after treatment with nitric oxide/peroxynitrite and redox agents.RESULTSBoth exogenous nitric oxide and peroxynitrite decreased cell growth and DNA synthesis of cultured rat aortic smooth muscle cells, but peroxynitrite-induced growth arrest was irreversible and associated with apoptosis and cytotoxicity. Inhibition of guanylate cyclase, PARP activity, mitogen-activated protein kinase, or bypass of ornithine decarboxylase did not reverse growth arrest by nitric oxide. The antioxidants N-acetylcysteine, ascorbate, and glutathione selectively reversed growth inhibition by nitric oxide but not by peroxynitrite. Antioxidants did not impair nitric oxide–induced cGMP generation in smooth muscle cells or nitric oxide–induced vasodilatation of isolated aortic rings. Nitric oxide treatment did not result in peroxynitrite formation and augmentation of superoxide levels did not induce peroxynitrite-like effects. Peroxynitrite-induced cytotoxicity and apoptosis were not reversed by antioxidants or PARP inhibition, because peroxynitrite activated PARP in J774 macrophages but failed to activate PARP in smooth muscle cells.CONCLUSIONSExogenous nitric oxide induces reversible cytostasis in smooth muscle cells by a redox-sensitive mechanism independent of peroxynitrite formation and distinct from the nitric oxide vasodilating mechanism. Peroxynitrite does not activate PARP selectively in smooth muscle cells and induces redox-independent smooth muscle cell cytotoxicity and apoptosis. Thus, the antiproliferative effects of nitric oxide and peroxynitrite on smooth muscle cells use divergent intracellular pathways with distinct redox sensitivities. These findings are relevant to the pathogenesis of vascular disease and the potential application of nitric oxide–based therapy for vascular disease.