ANGIOTENSIN-II STIMULATES NADH AND NADPH OXIDASE ACTIVITY IN CULTURED VASCULAR SMOOTH-MUSCLE CELLS
ANGIOTENSIN-II STIMULATES NADH AND NADPH OXIDASE ACTIVITY IN CULTURED VASCULAR SMOOTH-MUSCLE CELLS
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DOI：
10.1161/01.res.74.6.1141
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发表时间：
1994-06-01
影响因子：
20.1
通讯作者：
ALEXANDER, RW
中科院分区：
文献类型：
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作者：
GRIENDLING, KK;MINIERI, CA;ALEXANDER, RW
血管紧张素II（Ang II）在血管平滑肌细胞中的长期代谢效应所涉及的信号通路尚未完全了解，但包括可能影响氧化酶活性的分子的产生。我们研究了血管紧张素II刺激超氧阴离子形成的能力，并研究了负责其生产的氧化酶的身份。用血管紧张素II处理血管平滑肌细胞4 - 6小时，通过光泽精测定法检测到细胞内超氧阴离子形成增加2.7+/-0.4倍。这种超氧化物似乎是由NADPH和NADH氧化酶的激活引起的。在血管紧张素II作用4小时后，NADPH氧化酶活性从每毫克蛋白3.23+/-0.61增加到11.80+/-1.72 nmol O-2（-）/min，而NADH氧化酶活性从每毫克蛋白16.76+/-2.13增加到45.00+/-4.57 nmol O-2（-）/min。NADPH氧化酶活性被外源性磷脂酸和花生四烯酸刺激，并被特异性抑制剂二苯碘鎓部分抑制。花生四烯酸和亚油酸可增加NADH氧化酶的活性，对外源磷脂酸不敏感，高浓度的奎纳克林可抑制NADH氧化酶的活性。这两种氧化酶似乎都存在于质膜中，基于细胞分级分离后活性的迁移以及它们对线粒体毒物KCN的明显不敏感性。这些观察结果表明，血管紧张素II特异性激活酶系统，促进超氧化物的产生，并提高这些途径作为第二信使的长期反应，如肥大或增生的可能性。
The signaling pathways involved in the long-term metabolic effects of angiotensin II (Ang II) in vascular smooth muscle cells are incompletely understood but include the generation of molecules likely to affect oxidase activity. We examined the ability of Ang II to stimulate superoxide anion formation and investigated the identity of the oxidases responsible for its production. Treatment of vascular smooth muscle cells with Ang II for 4 to 6 hours caused a 2.7+/-0.4-fold increase in intracellular superoxide anion formation as detected by lucigenin assay. This superoxide appeared to result from activation of both the NADPH and NADH oxidases. NADPH oxidase activity increased from 3.23+/-0.61 to 11.80+/-1.72 nmol O-2(-)/min per milligram protein after 4 hours of Ang II, whereas NADH oxidase activity increased from 16.76+/-2.13 to 45.00+/-4.57 nmol O-2(-)/min per milligram protein. The NADPH oxidase activity was stimulated by exogenous phosphatidic and arachidonic acids and was partially inhibited by the specific inhibitor diphenylene iodinium. NADH oxidase activity was increased by arachidonic and linoleic acids, was insensitive to exogenous phosphatidic acid, and was inhibited by high concentrations of quinacrine. Both of these oxidases appear to reside in the plasma membrane, on the basis of migration of the activity after cellular fractionation and their apparent insensitivity to the mitochondrial poison KCN. These observations suggest that Ang II specifically activates enzyme systems that promote superoxide generation and raise the possibility that these pathways function as second messengers for long-term responses, such as hypertrophy or hyperplasia.