Protective Effect of Yiqi Fumai Injection(注射用益气复脉)on H9c2 Cell Damage Induced by TBHP Via AMPK/mTOR Pathway
Objective:To investigate the mechanism of Yiqi Fumai Injection(YQFM)in alleviating H9c2 cells injury induced by tert-butyl peroxide(TBHP)through regulating autophagy via AMPK/mTOR signaling pathway.Methods:H9c2 cells were cultured in vitro and divided into control group,TBHP group and TBHP+YQFM group.The survival rate of H9c2 cells was detected by CCK-8,and autophagy was detected by immuno-fluorescence The expression of autophagy-related proteins LC3Ⅱ/LC3Ⅰ and AMPK/mTOR pathway-related proteins were detected by Western blotting.Results:Precultured with 2,4,8,16 g/L YQFM could significantly restore the damage of H9c2 cells induced by TBHP.The immunofluorescence staining results showed that com-pared with the control group,the LC3 fluorescence intensity of cells was significantly increased in the TBHP group(P＜0.01);Compared with the TBHP group,the fluorescence intensity of LC3 in the TBHP+YQFM group was decreased,with a significant decrease observed in the 2 g/L YQFM group(P＜0.05).Western blot showed that the expressions of p-AMPK/AMPK,LC3-Ⅱ/LC3-1 and the ratio of p-mTOR/mTOR were sig-nificantly increased in TBHP group,while the ratio of p-AMPK/AMPK,LC3-Ⅱ/LC3-Ⅰ were significantly decreased in YQFM group,p-mTOR/mTOR ratio was significantly increased in YQFM group.Conclusion:Yiqi Fumai Injection can effectively antagonize the injury of H9c2 cells induced by TBHP,which may be related to the regulation of autophagy by AMPK/mTOR signaling pathway.
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