甘草次酸通过PI3K-AKT途径抑制H_2O_2所致大鼠心肌细胞氧化损伤

Glycyrrhetinic Acid Decreases H_2O_2-induced Oxidative Injury of H9C2 Cells through PI3K-AKT Pathway

目的:研究甘草次酸对H_2O_2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H_2O_2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;100μmol/L和200μmol/L的甘草次酸作用24 h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H_2O_2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 ceils 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 of apoptotic 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.