丹参多酚酸盐对过氧化氢所致乳鼠心肌细胞氧化应激损伤影响及其机制

Effects and Mechanism of Salvianolate on Oxidative Stress of Neonatal Rat Cadiocytes Induced by H_2O_2

目的:研究丹参多酚酸盐(Salvianolate)对过氧化氢(H2O2)所致乳鼠心肌细胞氧化应激损伤的影响及其作用机制。方法:于无菌环境下分离并培养乳鼠心肌细胞72 h后设空白对照组、H2O2组、丹参多酚酸盐(50、100、200 mg/L)+H2O2组,每组设8个复孔;给药干预24 h后,MTT法检测细胞存活率,检测细胞中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量,采用氧敏感荧光探针检测氧自由基(ROS)含量,检测培养液中心肌酶(AST、CPK、LDH);通过Flow Cytometry检测细胞凋亡状况并计算凋亡率,采用Western blot法检测NF-κB蛋白表达并进行半定量分析。结果:与H2O2组比较,丹参多酚酸盐(100、200 mg/L)+H2O2组细胞存活率显著升高;细胞中SOD、GSH-Px、CAT活性显著升高,ROS含量显著降低;培养液中AST、CPK、LDH和MDA含量显著降低,细胞凋亡率显著降低,NF-κB蛋白表达显著下调,上述差异均具有统计学意义(P<0.05,P<0.01)。结论:丹参多酚酸盐具有抑制H2O2所致乳鼠心肌细胞氧化应激损伤的作用;其作用机制可能与丹参多酚酸盐能够改善抗氧化酶活性,提高ROS清除能力,下调NF-κB蛋白表达,抑制细胞凋亡有关。

Objective：To investigate the effects and mechanism of salvianolate on oxidative stress of neonatal rat cadiocytes induced by H2O2. Methods：Cadiocytes of neonate rat was cultivated for 72 hours and divided into six groups：normal control group,H2O2 group,salvianolate（50,100,200 mg/L）＋H2O2groups（n=8）. Twelve hours after the drugs were given,the survival rate was detected by MTT;the activity of SOD,CAT,GSH-Px and the content of ROS in cardiomyocytes were determinted;the content of AST,CPK,LDH,MDA in culture medium were detected;the cardiomyocytes apoptosis was observed by flow cytometry and the apoptosis rate was calculated;the expression of NF-κB protein were detected by Western blot and semi-quantitative analysized. Results：Compared with H2O2 group,the survival rate in salvianolate（100,200 mg/L）＋H2O2 groups was significantly increased;the activity of SOD,GSH-Px,CAT in cardiomyocytes were significantly increased and the content of ROS were significantly decreased;the content of AST,CPK,LDH and MDA in culture medium were significantly decreased;the cardiomyocytes apoptosis were improved and the apoptosis rate were significantly decreased,the expression of NF-κB protein was significantly down-regulated,all of the difference were significant（P〈0.05,P〈0.01）. Conclusion：salvianolate had inhibitive effects on oxidative stress of neonatal rat cadiocytes induced by H2O2,which perhaps related to its effects of improving the activity of antioxidase,enhancing the free radical scavenging ability,down-regulating the expression of NF-κB protein,depressing apoptosis.