丹参多酚酸盐对过氧化氢所致乳鼠心肌细胞损伤保护作用的研究

Protective Effect of Salvianolate on Cardiomyocytes Damage of Neonatal Rats Induced by H_2O_2

目的:研究丹参多酚酸盐(Salvianolate)对过氧化氢(H_2O_2)所致乳鼠心肌细胞损伤的保护作用及其机制。方法:分离并培养乳鼠心肌细胞72 h后随机分为空白对照组、H_2O_2组、丹参多酚酸盐(50、100和200 mg/L)+H_2O_2组,每组设8个复孔;给药干预24 h后,通过光学显微镜观察细胞形态,MTT法测定细胞存活率;检测细胞中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量,测定培养液中谷草转氨酶(AST)、磷酸肌酸激酶(CPK)、乳酸脱氢酶(LDH)含量;ELISA法测定培养液中TNF-α、IL-1、IL-6含量;通过Flow Cytometry检测细胞凋亡状况并计算细胞凋亡率。结果:与H_2O_2组比较,丹参多酚酸盐各干预组乳鼠心肌细胞生存状态呈不同程度好转,其中以丹参多酚酸盐(200mg/L)+H_2O_2组效果最为显著;丹参多酚酸盐(100和200 mg/L)+H_2O_2组细胞存活率显著升高,SOD、CAT活性显著升高且MDA含量显著降低,培养液中AST、CPK、LDH含量显著降低,TNF-α、IL-1、IL-6水平均显著降低;丹参多酚酸盐各干预组心肌细胞凋亡状况呈不同程度改善,其中丹参多酚酸盐(100和200 mg/L)+H_2O_2组凋亡率显著降低,上述差异均具有统计学意义(P<0.05,P<0.01)。结论:丹参多酚酸盐能够通过改善细胞生存状态、提高细胞存活率、降低氧化应激损伤、抑制炎症反应、抑制细胞凋亡而对H_2O_2所致乳鼠心肌细胞损伤起到保护作用。

Objective： To investigate the protective effect of Salvianolate on cardiomyocytes damage of neonatal rats induced by H_2O_2. Methods： Cardiomyocytes damage of neonatal rats was induced for 72 hours and they were divided into the following groups： normal control group, H_2O_2 group, Salvianolate（ 50, 100,200 mg / L） ＋ H_2O_2groups（ n = 8）. Twelve hours after the drugs were given,and the morphology changes were observed; the survival rate was detected by MTT; the activities of SOD,CAT and the content of MDA in cardiomyocytes were determinted; the activities of AST,CPK,LDH in culture medium were detected; the levels of TNF- α,IL- 1 and IL- 6 were detected by ELISA; the cardiomyocytes apoptosis was observed by flow cytometry and the apoptosis rate was calculated. Results： Compared with H_2O_2 group,the morphology in Salvianolate groups was significantly improved; the survival rates in Salvianolate（ 100,200 mg / L） ＋ H_2O_2 groups were significantly increased; the activities of SOD,CAT in cardiomyocytes were significantly increased and the contents of MDA were significantly decreased; the activities of AST,CPK,LDH in culture medium were significantly decreased; the levels of TNF- α,IL- 1,IL- 6 were significantly decreased; the cardiomyocytes apoptosis was improved and the apoptosis rates were significantly decreased. All of the differences were significant（ P〈0. 05,P〈0. 01）. Conclusion： Salvianolate can effectively improve morphology,improve the activity of antioxidase,depress oxidative stress and inhibit inflammatory response,suggesting that Salvianolate has a protective effect on cardiomyocytes damage of neonatal rats induced by H_2O_2.