白藜芦醇对高脂所致心肌细胞氧化应激损伤的保护作用及其与AMPK/mTORC1/p70S6K信号通路的相关性

Protective effect of resveratrol on oxidative stress injury of cardiomyocytes induced by high fat and its correlation with AMPK/mTORC1/p70S6K signaling pathway

目的探讨白藜芦醇对脂毒性心肌细胞损伤的保护作用及其机制。方法棕榈酸(palmitic acid,PA)刺激H9c2细胞,白藜芦醇预处理后,MTT检测细胞增殖情况;免疫荧光及流式细胞法检测活性氧(reactive oxygen species,ROS)水平;用试剂盒检测丙二醛(malondialdehyde,MDA)及超氧化物歧化酶(superoxide dismutase,SOD)水平;免疫印迹法检测AMPK/m TORC1/p70S6K通路蛋白以及凋亡蛋白表达水平。结果 MTT结果示,PA(0. 4 mmol·L-1)刺激24、48 h时,心肌细胞存活率出现明显下降;免疫荧光法及流式细胞法显示,与对照组相比,PA(0. 4 mmol·L-1)刺激24 h时,ROS及MDA水平明显升高,SOD活性明显降低;免疫印迹法显示,与对照组相比,PA(0. 4 mmol·L-1)刺激24 h时,p-AMPK蛋白及Bcl-2蛋白表达降低,p-mTORC1、p-p70S6K、Bax和cleaved caspase-3蛋白表达增高。白藜芦醇预处理,可以明显逆转上述变化。结论白藜芦醇可以明显抵抗高脂所致的心肌细胞氧化应激损伤,可能与AMPK/m TORC1/p70S6K信号通路调控相关。

Aim To investigate the protective effect of resveratrol on oxidative stress injury of cardiomyocytes induced by high fat and its correlation with AMPK/mTORC1/p70S6K signaling pathway.Methods The model of cardiomyocyte lipotoxicity injury was established by palmitic acid (PA).Resveratrol pretreatment and MTT assay were used to detect cell proliferation.The expression of reactive oxygen species (ROS) was detected by fluorescence and flow cytometry, and the levels of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were detected by kit.The protein expression of AMPK/mTORC1/P70S6K signaling pathway and apoptosis related proteins were determined by Western blot.Results MTT results showed that the cell viability decreased significantly at 0.4 mmol·L^-1 and24 h-48 h stimulation time, respectively.Fluorescence and flow cytometry showed that the levels of ROS and MDA increased significantly and SOD activity decreased significantly when PA (0.4 mmol·L^-1 ) stimulated cells for24 h compared with control group.Western blot showed that the expression of p-AMPK and Bcl-2 decreased and the expression of p- mTORC1, p-p70S6K, Bax and cleaved caspase-3 increased when PA (0.4 mmol·L^-1 ) stimulated cells for24 h compared with control group.Resveratrol pretreatment could significantly reverse the above changes.Conclusions Resveratrol can significantly inhibit the apoptosis of cardiomyocytes induced by high fat.Its mechanism may be related to the regulation of AMPK/mTORC1/p70S6K signaling pathway.