Significance Of Apoptosis And Autophagy In Myocardial Cell In Jury Resulting Of Oxidative Stress

ObjectiveDetecting the expression of autophagy factor for Beclin-1, LC3-Ⅱ, ULK1 and apoptosis factor for Bax, caspase in myocardial cell injury model that 3% hydrogen peroxide intervent it. To investigate the effect of autophagy in myocardial cell injury resulting of oxidative stress and discuss the correlation of apoptosis and autophagy.MethodsHCM-C myocardial cell were trained in Incubator of 37℃,5%C02 and passaged according to the growth condition of cells. shRNA targeting Beclin-1 were chemically synthesized and transfected into HCM-C cell lines in vitro. The Beclin-1 was activated by rapamycin. A cell model of myocardial cell injury was established by 3%H2O2 interventing HCM-C cardiomyocytes for 6h. The cells were divided into four groups: including blank control group and experimental groups H2O2 group, Beclin-1 shRNA +H2O2 group and rapamycin +H2O2 group).The mRNA and protein levels of Beclinl、LC3-Ⅱ、ULK1、Bax、caspase3 expression were detected by RT-PCR and Western-blot. Apoptosis were determined by flow cytometry (FCM). CCK-8 assay was used to assess the cell survival rate.Results1.A final concentration of 200umol/L was established by the 3% H2O2 2.3ul was added 10% DMEM (containing fetal bovine serum) culture medium of 10ml.Which intervented HCM-C myocardial cell for 6h, to establish the most appropriate model of myocardial cell injury.2.The mRNA and protein levels of Beclin-1、LC3-Ⅱ、Bax、caspase-3、ULK1 was increased in myocardial cell injury group compared with control group.3.The apoptotic rate of the Beclin-1 shRNA +H2O2 group was higher than the H2O2 group, whereas the rapamycin +H2O2 group was lower than the H2O2 group.4.The survival rate of the Beclin-1 shRNA +H2O2 group was higher than the H2O2 group, whereas the rapamycin +H2O2 group was lower than the H2O2 group.Conclusion1.During myocardial cell injury induced by H2O2, autophagy and apoptosis are enhanced.2.In the process of oxidative stress by H2O2, autophagy and apoptosis are mutual antagonism and coexist in the cell death process. autophagy and apoptosis program will be started up. During this process, autophagy plays a major role.3. Under oxidative stress, autophagy exists as a disadvantage in myocardial cell injury.myocardial cells occur autophagic death in order to adapt to environmental changes, when autophagy was inhibited, the cell survival rate increased.