橙皮素对H2O2诱导H9c2心肌细胞凋亡的抑制效应研究

Effect of hesperetin in inhibition on H9c2 cardiomyocytes apoptosis induced by H2O2

目的 探讨橙皮素（HES）对H2O2诱导的H9c2心肌细胞凋亡的影响.方法 采用H2O2建立H9c2心肌细胞氧化应激损伤模型.实验分为4组：正常对照组（Control组）、H2O2损伤组（H2O2组）、单纯橙皮素处理组（HES组）、橙皮素预处理＋H2O2组（HES＋H2O2组）.H2O2（400μM）处理2 h建立心肌细胞氧化应激损伤模型,HES＋H2O2组于建模前1 h加入40μM橙皮素.采用CCK-8法确定H9c2细胞活性,DCFH-DA探针检测细胞活性氧簇（ROS）水平,流式细胞术检测心肌细胞凋亡,分光光度计检测Caspase-3活性.结果橙皮素预处理可明显改善H2O2诱导的H9c2心肌细胞活性降低,且浓度为40μM时保护作用最明显;给予40μM橙皮素预处理后ROS的产生明显减少,Caspase-3活性显著下降,心肌细胞凋亡率为（28.32±2.12）%,明显低于H2O2组（50.33±2.56）%（P〈0.05）.结论 橙皮素对氧化应激诱导的心肌细胞凋亡具有抑制效应.

Objective To study the protective effects of hesperetin pretreatment on H9c2 cardiomyocytes after oxidative stress injured by hydrogen peroxide （H2O2）. Methods H9c2 cells were treated with H2O2 to estab-lish a myocardial injury model. The experiment was divided into four groups：control group;model group（H2O2）;hesperetin pretreated group （HES）and hesperetin pretreated＋model group （HES＋H2O2）. H9c2 cardiomyocytes in model group and hesperetin pretreated＋model group were exposed H2O2 to for 2 h for establishing the model of ox-idative stress injury. Hesperetin pretreated＋model group were pretreated with hesperetin for 1 h befor subjected H2O2 to exposure. After the indicated treatment,the cell viability was assessd with CCK-8 kit：ROS levels were detected by DCFH-DA probe; cell apoptosis were analyzed by d flow cytometry and Caspase-3 activities were also detected. Results The improvement of hesperetin pretreatment on H9c2 cells activity decrease which induced by H2O2,and this phenomenon was observed obviously on the concentration of 40 μM. On the concentration of 40 μM, hesperetin pretreatment could reduce ROS production,increase Caspase-3 activity and the apoptosis of myocardial cells （28.32±2.12）% was significantly decreased in the H2O2 group （50.33±2.56）%（P〈0.05）. Conclusion Hes-peretin pretreatment can provide significantly cardioprotective effects against oxidative stress injury.