基于AMPK信号通路探讨虫草素对过氧化氢致心肌细胞损伤的保护作用

Research of Protective Effect of Cordycepin on Myocardial Oxidative Damage through AMPK Signaling Pathway

目的:观察虫草素对过氧化氢致心肌细胞损伤的保护作用并探讨其与AMPK信号通路的相关性。方法:培养H9C2心肌细胞分为正常组(Control)、氧化损伤组(oxidative damage group,ODG)、虫草素组(Cordycepin);其中氧化损伤组H9C2心肌细胞通过双氧水构建氧化损伤模型,虫草素组在氧化损伤前使用虫草素预处理;CCK8实验测定各组细胞活力值,流式细胞仪检测各组细胞ROS的表达,Western blot检测各组细胞AMPK信号通路相关蛋白的表达。结果:与Control组比较,ODG组与Cordycepin组细胞活力值均明显下降(均P<0.05),细胞内ROS表达量均明显上升(均P<0.05);与ODG组比较,Cordycepin预处理后,细胞活力值明显提升(P<0.05),细胞内ROS表达量明显降低(P<0.05),p-AMKP/AMPK的表达明显上调(P<0.05)。结论:虫草素预处理可有效发挥对过氧化氢致心肌细胞氧化损伤的保护作用,其机制可能与AMPK信号通路的激活相关。

Objective:To research the protective effect of Cordycepin on oxidative damage of cardiomyocytes induced by hydrogen peroxide and its correlation with AMPK signal pathway.Methods:H9C2 cardiomyocytes were cultured and divided into three groups:Control Group,Oxidative Damage Group(ODG)and Cordycepin Group;H9C2 myocardial cells in oxidative damage group were cultured with hydrogen peroxide to establish oxidative damage model,and Cordycepin group was pretreated with Cordycepin on the basis of oxidative damage group;The optimal protective concentration of Cordycepin against oxidative damage was determined by CCK8 experiment,the expression of ROS in each group was detected by flow cytometry,and the expressions of AMPK signal pathway related proteins in each group were detected by Western blot.Results:Compared with control group,the cell viability of ODG group and Cordycepin group decreased significantly(all P<0.05),and the expression of intracellular ROS increased significantly(all P<0.05).Compared with ODG group,after Cordycepin pretreatment,the cell viability increased significantly(P<0.05),the expression of intracellular ROS decreased significantly(P<0.05),and the expression of p-AMPK/AMPK increased significantly(P<0.05).Conclusion:Cordycepin pretreatment can effectively protected myocardial cells from oxidative damage induced by hydrogen peroxide,and its mechanism may be related to the activation of AMPK signaling pathway.