Regulation on autophagy with tanshinone ⅡA for anti-oxidative stress damage of endothelial cells through PI3K/Akt/mTOR pathway
Objective To investigate the protective and mechanism of tanshinone Ⅱ A (TS Ⅱ A) in oxidative stress damage of endothelial cells induced by oxidized low-density lipoprotein (ox-LDL) based on PI3K/Akt/mTOR autophagy pathway.Methods EA.hy926 cells were cultured in vitro,and then randomly divided into normal group,model group,ox-LDL + TS Ⅱ A group,ox-LDL + TS Ⅱ A + LY294002 group,TS Ⅱ A group and LY294002 group.The content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) were detected by using chromatoptometry,autophagy status was observed by using FITC-LC3 fluorescence microscope and Naolive 3D Cell Explorer Real-time Microscopic Imaging System,and meanwhile,expressions of autophagy-related proteins and PI3K/Akt/mTOR pathway proteins were detected by using Western Blotting.Results Compared with normal group,MDA content increased,SOD activity decreased and content of LC3-Ⅰ/LC3-Ⅱ protein increased in model group (P ＜0.01),and after TS Ⅱ A intervention,MDA content decreased,SOD activity increased and content of LC3-Ⅰ/LC3-Ⅱ protein increased (P ＜ 0.01).Compared with normal group,protein expressions of PI3K,p-Akt and p-mTOR decreased significantly in model group (P ＜ 0.05 or P ＜ 0.01).Compared with model group,protein expressions of PI3K,p-Akt and p-mTOR decreased significantly after TS ⅡA intervention (P ＜0.05 or P ＜0.01).Compared with ox-LDL + TS Ⅱ A group,content of LC3-Ⅰ/LC3-Ⅱ protein decreased and autophagy level decreased (P ＜ 0.01) after applying LY294002,an inhibitor of PI3K.Conclusion Tanshinone ⅡA may improve autophagy through regulating PI3K/Akt/mTOR pathway to relieve the oxidative stress damage of EA.Hy926 cells and prevent atherosclerosis.
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