Aldosterone stimulates reactive oxygen species production through activation of NADPH oxidase in rat mesangial cells
Aldosterone stimulates reactive oxygen species production through activation of NADPH oxidase in rat mesangial cells
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DOI：
10.1681/asn.2005040390
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发表时间：
2005-10-01
影响因子：
13.6
通讯作者：
Nishiyama, A
中科院分区：
文献类型：
--
作者：
Miyata, K;Rahman, M;Nishiyama, A
最近的研究表明，长期接受醛固酮和盐治疗的大鼠肾小球系膜损伤与肾皮质活性氧 (ROS) 水平升高有关。本研究旨在确定培养的大鼠系膜细胞 (RMC) 中醛固酮诱导 ROS 产生的机制。氧化荧光二氢乙锭用于评估完整细胞中超氧阴离子（O-2（-））的细胞内产生。光泽精衍生的化学发光测定用于测定 NADPH 氧化酶活性。醛固酮（1 至 100 nmol/L）以剂量依赖性方式增强二氢乙锭的染色，在 RMC 中在 3 It 处出现峰值。醛固酮（100 nmol/L，持续 3 小时）还显着增加了 NADPH 氧化酶活性，从 232 +/- 18 增加到 346 +/- 30 cpm/5 X 10(4) 细胞。免疫印迹数据显示醛固酮（100 nmol/L，持续 3 小时）使膜组分中的 p47phox 和 p67phox 蛋白水平分别增加约 2.1 和 2.3 倍。另一方面，RMC 中 NADPH 氧化酶膜成分 p22phox、Nox-1 和 Nox-4 的 mRNA 表达不被醛固酮改变（持续 3 至 12 小时）。与选择性盐皮质激素受体（MR）拮抗剂依普利酮（10 μmol/L）预孵育，显着减弱醛固酮诱导的O-2(-)产生、NADPH氧化酶激活以及p47phox和p67phox的膜易位。这些结果表明，醛固酮诱导的 ROS 生成与 RMC 中 p47phox 和 p67phox 的 MR 介导的膜易位与 NAPDH 氧化酶激活相关。醛固酮的这些细胞作用可能在肾小球系膜损伤的发病机制中发挥作用。
It has recently been shown that glomerular mesangial injury is associated with increases in renal cortical reactive oxygen species (ROS) levels in rats treated chronically with aldosterone and salt. This study was conducted to determine the mechanisms responsible for aldosterone-induced ROS production in cultured rat mesangial cells (RMC). Oxidative fluorescent dihydroethidium was used to evaluate intracellular production of superoxide anion (O-2(-)) in intact cells. The lucigenin-derived chemiluminescence assay was used to determine NADPH oxidase activity. The staining of dihydroethidium was increased in a dose-dependent manner by aldosterone (1 to 100 nmol/L) with a peak at 3 It in RMC. Aldosterone (100 nmol/L for 3 h) also significantly increased NADPH oxidase activity from 232 +/- 18 to 346 +/- 30 cpm/5 X 10(4) cells. Immunoblotting data showed that aldosterone (100 nmol/L for 3 h) increased p47phox and p67phox protein levels in the membrane fraction by approximately 2.1- and 2.3-fold, respectively. On the other hand, mRNA expression of NADPH oxidase membrane components, p22phox, Nox-1, and Nox-4, were not altered by aldosterone (for 3 to 12 h) in RMC. Pre-incubation with the selective mineralocorticoid receptor (MR) antagonist, eplerenone (10 mu mol/L), significantly attenuated aldosterone-induced O-2(-) production, NADPH oxidase activation and membranous translocation of p47phox and p67phox. These results suggest that aldosterone-induced ROS generation is associated with NAPDH oxidase activation through MR-mediated membranous translocation of p47phox and p67phox in RMC. These cellular actions of aldosterone may play a role in the pathogenesis of glomerular mesangial injury.