Genetic demonstration of p47phox-dependent superoxide anion production in murine vascular smooth muscle cells
Genetic demonstration of p47phox-dependent superoxide anion production in murine vascular smooth muscle cells
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DOI：
10.1161/01.cir.104.1.79
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发表时间：
2001-07-03
期刊：
影响因子：
37.8
通讯作者：
Leto, TL
中科院分区：
文献类型：
--
作者：
Lavigne, MC;Malech, HL;Leto, TL
背景-以前的研究提供了证据表明，一种与吞噬细胞NADPH氧化酶相关的酶在血管壁中产生超氧化物。然而，这些数据被观察到的NADPH和NADH都是血管细胞中超氧化物产生的底物所混淆。为了阐明这个问题，我们比较了野生型血管平滑肌细胞（VSMCs）产生超氧化物的能力，（p47 phox（+/+）;吞噬细胞氧化酶）小鼠与来自缺乏p47 phox的小鼠的那些（p47phox（-/-）;“敲除”），吞噬细胞NADPH氧化酶的必需组分，方法nod结果VSMCs来源于p47 phox（+/+）或p47 phox（-/-）小鼠的主动脉组织块。p47 phox（+/+）小鼠的VSMCs在佛波醇肉豆蔻酸酯刺激后产生超氧化物，而p47 phox（-/-）小鼠的VSMCs则没有。与此一致，p47 phox仅在p47 phox（+/+）VSMCs中检测到。在血管紧张素II或血小板衍生生长因子-BE（PDGF-BB）刺激后，p47 phox转导的p47 phox（-/-）VSMCs产生显著水平的超氧化物，但没有增强的绿色荧光蛋白转导的p47 phox（-/-）VSMCs产生显著水平的超氧化物。p47 phox转导的p47 phox（-/-）细胞中重组p47 phox的表达增强与这些cells. Conclusions超氧化物的产生相关，这些数据提供了直接的功能证据，即需要p47 phox组分的氧化酶介导血管壁中VSMCs的超氧化物释放，以响应血管紧张素II或PDGF-BB。
Background-Previous investigations provide evidence that an enzyme related to the phagocyte NADPH oxidase produces superoxide in the blood vessel wall. These data, however, are confounded by observations that both NADPH and NADH serve as substrates for superoxide production in vascular cells. To clarify this issue, we compared the superoxide-generating capabilities of vascular smooth muscle cells (VSMCs) derived from wild-type (p47phox(+/+); phagocyte oxidase) mice with those from mice that lack p47phox (p47phox(-/-); "knockout"), an essential component of the phagocyte NADPH oxidase,Methods nod Results-VSMCs were derived from aortic explants harvested from p47phox(+/+) or p47phox(-/-) mice. VSMCs from p47phox(+/+) but not those from p47phox(-/-) mice produced superoxide after stimulation by phorbol myristate acetate. Consistent with this, p47phox was detected only in p47phox(+/+) VSMCs. p47phox-transduced p47phox(-/-) but not enhanced green fluorescent protein-transduced p47phox(-/-) VSMCs generated significant levels of superoxide after stimulation by angiotensin II or platelet-derived growth factor-BE (PDGF-BB). Enhanced expression of recombinant p47phox in p47phox-transduced p47phox(-/-) cells correlated with superoxide production in these cells.Conclusions-These data provide direct functional proof that an oxidase requiring the p47phox component mediates superoxide release from VSMCs in the blood vessel wall in response to angiotensin II or PDGF-BB.