冠心苏合胶囊大鼠含药血清抗氧化损伤致心肌凋亡机制研究

Mechanism of Guanxin Suhe-containing serum against apoptosis of cardiomyocytes induced by hydrogen peroxide

目的探讨冠心苏合胶囊大鼠含药血清对氧化损伤心肌细胞凋亡的作用及其机制。方法从服用冠心苏合胶囊的大鼠腹主动脉采集含药血清,对原代乳鼠心肌细胞H2O2氧化损伤模型进行干预。用MTT法检测细胞存活率;Ho-echst33258荧光染色法观察药物作用后心肌细胞形态学变化;caspase-3活性测定方法探讨冠心苏合含药血清抗凋亡途径;Western-blot方法测定冠心苏合含药血清对Bcl-2及Bax蛋白表达的影响。结果 MTT法作用结果显示,将原始收集血清稀释为相当于灌胃0.4、0.6、0.8 g/kg冠心苏合胶囊时,相对于空白血清组和H2O2单纯损伤模型组,心肌细胞有较高的存活率(P<0.01),且呈现一定的剂量依赖性;Hoechst33258荧光染色结果显示,与模型组相比,含药血清组随着药物浓度升高细胞核固缩凝聚减轻,碎片依次减少;与模型组相比,caspase-3活性随含药血清浓度增高而降低,冠心苏合给药组能够下调Bax水平,上调Bcl-2水平,且呈现剂量依赖性。结论药物冠心苏合能够提高H2O2诱导的原代乳鼠心肌细胞存活率,降低caspase-3活性,减轻细胞核固缩和聚集,减少细胞核碎片,且能够降低Bax表达水平,提高Bcl-2表达水平。

AIM To study the mechanism of Guanxin Suhe-containing serum against apoptosis of cardiomyocytes induced by hydrogen peroxide. METHODS The drug-containing serum collected and separated from abdominal aorta of SD rat fed by Guanxin Suhe Capsules was used to treat the primary cultured cardiomyocytes injured by hydrogen peroxide. The cardiomyocyte viability was determined by MTY method. Morphological changes of cardiomyocytes were observed by fluorescence microscope. The anti-apoptotic effect was detected by the activity of caspase-3. The protein expressions of Bcl-2 and Bax were semi-quantified by Western-blot method. RESULTS The cardiomyocyte viability was elevated （P 〈 0. 01 ） after being treated with different concentrations of drug-contai- ning serum （0. 2, 0.4, 0. 8 g/kg, diluted from Guanxin Suhe Capsules）. Typical apoptotic features of the cardio- myocytes such as membrane blebbing, cell shrinkage and detachment, and nucleus condensation and fragmentation were dose-dependently improved after being treated with drug-containing serum. The activity of caspase-3 decreased with increasing the concentration of drug. Western blot approach showed that the Bcl-2 level increased meanwhile Bax decreased. CONCLUSION Guanxin Suhe-containing serum could increase the viability of the primary cultured cardiomyocytes in hydrogen peroxide induced injury. It could also decrease the activity of caspase-3, improve nucleus condensation and fragmentation, increase Bcl-2 level and decrease the Bax level.