苦参素对H2O2诱导乳鼠心肌细胞氧化应激损伤保护作用的研究  

Oxymatrine Protects Cadiocytes of Neonatal Rat Against H_2O_2-induced Oxidative Stress Injury

作　　者：王雪芬[1] 陈树杰[2] 王磊[2] 

机构地区：[1]邯郸市第二医院内一科,056001 [2]邯郸市中心医院,056001

出　　处：《医学研究杂志》2016年第8期85-89,共5页Journal of Medical Research

基　　金：河北省卫生厅重点科技研究计划项目(20130359)

摘　　要：目的研究苦参素(oxymatrine,OMT)对H_2O_2诱导乳鼠心肌细胞氧化应激损伤的保护作用。方法无菌环境下分离并培养乳鼠心肌细胞72h后随机分为空白对照组、H_2O_2(200μmol/L)干预组、OMT(20、40、60μmol/L)+H_2O_2(200μmol/L)干预组,每组设10个复孔。药物干预12h后,MTT法检测细胞存活率,采用氧敏感荧光探针DCFH-DA检测氧自由基(ROS);测定细胞中抗氧化酶(SOD、GSH-Px、CAT)活性和丙二醛(MDA)含量,检测细胞培养液中心肌酶(AST、CPK、LDH)含量,流式细胞术(flow cytometry,FCM)检测细胞凋亡并计算凋亡率,Western blot法检测细胞中caspase-3、NF-κB蛋白表达并进行半定量分析。结果与H_2O_2(200μmol/L)干预组比较,OMT(40、60μmol/L)干预12h能够显著提高H_2O_2诱导损伤乳鼠心肌细胞存活率,降低ROS含量,改善SOD、GSH-Px、CAT活性并降低MDA含量,降低培养基中AST、CPK、LDH含量,降低细胞凋亡率,降低caspase-3、NF-κB蛋白表达,差异均有统计学意义(P<0.05,P<0.01)。结论苦参素可能通过改善抗氧化酶活性而提高氧自由基清除能力、抑制促凋亡蛋白表达而降低细胞凋亡,对H_2O_2诱导乳鼠心肌细胞氧化应激损伤起到保护作用。Objective To investigate the protective effects of Oxymatrine （OMT） on cardiomyocytes of neonatal rat against H2O2 - induced oxidative stress injury. Methods Cadiocytes of neonate rat were cultivated for 72 hours and divided into six groups ： normal control group, H2O2 （200μmol/L） group, OMT（20, 40, 60μmol/L） ＋ H2O2 （200μmol/L） groups（n = 10）. Twelve hours after the drugs were given, the survival rate was detected by MTT, the content of ROS in cardiomyocytes were determinted by oxygen - sensitive fluores- cent probe, the activity of SOD, GSH -Px, CAT and the content of MDA were determinted, the content of AST, CPK, LDH in culture medium were detected, the cardiomyocytes apoptosis was observed by flow cytometry and the apoptosis rate was calculated, the expression of caspase- 3, NF - κB proten were determined by Western blot and Semi - quantitative analysized. Results Compared with H202 （200μmol/L） group, the survival rate of OMT（40, 60μmol/L） ＋ H2O2 （200 μmol/L） groups were significantly increased, the content of ROS in cardiomyocytes were significantly decreased, the activity of SOD, GSH - Px, CAT were significantly increased and the content of MDA was significantly decreased, the content of AST, CPK, LDH in culture medium were significantly decreased, the apoptosis rate were significantly decreased, the expression of caspase - 3, NF - κB protein was significantly decreased, and all of the difference above was significant（P 〈 0. 05, P 〈 0.01 ）. Conclusion OMT protectes neonatal rat eardiomyocytes from H2O2 -induced oxidative stress injury probably through improving the activity of antioxidase to increase the oxygen radical scavenging, regulating the expression of apoptosis -related protein to inhibite apoptosis.

关 键 词：苦参素 H2O2 心肌细胞保护 

分 类 号：R393[医药卫生—基础医学]