目的:研究甘草次酸对H2O2所致大鼠心肌细胞氧化损伤的影响及其可能机制.方法:采用H2O2处理建立H9c2大鼠心肌细胞氧化损伤模型后,比较模型中ROS生成和细胞凋亡比例,使用不同浓度的甘草次酸孵育H9c2细胞24、48h后,通过流式细胞仪检测ROS的生成量,Annexin V-FITC/PI双标记流式细胞术检测凋亡率,蛋白质印迹法检测检PI3K、AKT1、p-AKT蛋白的表达.结果:H9c2大鼠心肌细胞氧化损伤模型中ROS生成和细胞凋亡率分别为(49.33±3.23)％和(33.89±1.45)％,与对照组相比有显著差异;100 μmol/L和200 μmol/L的甘草次酸作用24h后,H9c2大鼠心肌细胞氧化损伤模型中表达ROS细胞的比例(35.39±1.24)％和(30.46±0.95)％,凋亡细胞比例分别为(29.47± 3.15)％和(23.17± 1.46)％,当作用48 h后,H9c2大鼠心肌细胞氧化损伤模型中表达ROS细胞的比例(42.67± 1.89)％和(35.49±1.63)％,凋亡细胞比例分别为(40.22± 3.06)％和(35.26± 2.78)％,与对照组相比有显著性差异;使用渥曼青霉素后,各甘草次酸组的与对照组无显著性差异.结论:甘草次酸可能通过PI3K-AKT途径抑制H2O2所致大鼠心肌细胞氧化损伤.%Objective:To study the effect and mechanism of glycyrrhetinic acid on the oxidative injury of rat myocardial cells.Methods:Rat myocardial H9c2 cells were treated by H2O2 to induce oxidative injury.The ROS generation and apoptosis were detected.Different concentrations of glycyrrhetinic acid were used to treat H2O2-induced H9c2 cells for 24 h and 48 h.of the ROS generation in H9c2 cells was detected by flow cytometry;proportion of apoptotic cells was detected by flow cytometry with Annexin V/PI double marker staining;PI3K,AKT1,p-AKT protein expressions were detected by Western blotting.Results:The ROS generation and cell apoptotic rate in H2O2-induced H9c2 cells were (49.33± 3.23)％ and (33.89± 1.45)％ respectively,which were significantly higher than those of the control group.100 μmol/L and 200 μmol/L glycyrrhetinic acid obviously decreased the ROS generation[(35.39± 1.24)％ and (30.46± 0.95)％],ratios ofapoptotic cell [(29.47± 3.15)％ and (23.17± 1.46)％] in H9c2 cells treated by H2O2 for 24 hours;100μmol/L and 200μmol/L glycyrrhetinic acid obviously decreased the ROS generation [(42.67± 1.89)％ and (35.49± 1.63)％],ratios ofapoptotic cells [(40.22± 3.06)％ and (35.26 ± 2.78)％] in H9c2 cells treated by H2O2 for 48 hours;wortmannin showed similar effect with glycyrrhetinic acid.Conclusion:Glycyrrhetinic acid could decrease the generation of ROS and apoptosis in H2O2-induced oxidative injury of H9c2 cells through PI3K-AKT pathway.
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