螺内酯对醛固酮诱导的大鼠肾小球系膜细胞氧化应激的影响

目的 观察螺内酯对醛固酮诱导的大鼠肾小球系膜细胞(mesangial cells,MCs)氧化应激的影响,探讨其肾脏保护机制.方法 体外培养大鼠MCs,设正常对照组、醛固酮(10-7 mol/L)刺激组、不同浓度螺内酯(10-7、10-8、10-9mol/L)干预组.以2',7'-二氯双氢荧光素二乙酸酯(2',7'-dichlorofluorescein diacetate,DCFH-DA)为细胞内H2O2的荧光探针标记细胞,采用流式细胞术检测系膜细胞内活性氧(reactive oxygen species,ROS)水平；采用比色法检测培养细胞上清液中丙二醛(malondialdehyde,MDA)的含量和超氧化物歧化酶(superoxide dismutase,SOD)活性.结果 与正常对照组比较,醛固酮刺激系膜细胞48 h后,细胞内ROS水平明显增加,上清液中MDA含量增高、SOD活性下降,差异均有统计学意义(均有P＜0.05).螺内酯干预可明显抑制醛固酮刺激下的系膜细胞的上述反应,差异均有统计学意义(均有P＜0.05),且呈浓度依赖性.结论 螺内酯可呈浓度依赖性地抑制醛固酮诱导的大鼠MCs氧化应激,该作用可能是其肾脏保护机制之一.

Effects of spironolactone on aldosterone mediated oxidative stress in cultured rat glomerular mesangial cells

Objective To observe the effects of spironolactone on aldosterone mediated oxidative stress,in order to explore its mechanisms of reno-protection.Methods Rat glomerular mesangial cells were cultured with normal glucose concentration,aldosterone（10-7mol/L） and different concentrations of spironolactone（10-7,10-8,10-9mol/L）.The levels of intracellular reactive oxygen species（ROS） were measured by flow cytometry,which used 2＇,7＇-dichlorofluorescein diacetate（DCFH-DA） as a reactive oxygen species fluorescent dye.The levels of malondialdehyde（MDA） and superoxide dismutase（SOD） were determined by colorimetry method.Results Compared with normal glucose group,the intracellular reactive oxygen species content in mesangial cells（P0.05） and malondialdehyde levels（P0.05） in the supernatants increased significantly in aldosterone group,while spironolactone inhibited those changes in a dose-dependent manner（P0.05）,and the superoxide dismutase activity in the supernatant（P0.05） displayed the opposite changes.Conclusions Spironolactone can suppress aldosterone mediated oxidative stress in glomerular mesangial cells with a dose-dependent manner,which may partly contribute to its reno-protection.