Effects of transfection with HACE1 gene overexpression or knockdown viral plasmids on viability,apoptosis,and HO-1 expression in cardiomyocytes with acute oxidative stress injury
Objective To investigate the effects of transfection with HACE1 gene overexpression or knockdown viral plasmids on cell viability,apoptosis,and heme oxygenase-1(HO-1)expression in cardiomyocytes H9C2 with acute oxi-dative stress injury.Methods H9C2 cardiomyocytes were divided into three groups:control group,HACE1 overex-pression(HACE1-OE)group,and HACE1 knockdown(shRNA-HACE1)group.The cells in the HACE1-OE group were transfected with the HACE1 overexpression viral plasmid,and cells in the shRNA-HACE1 group were transfected with HACE1 knockdown viral plasmid(shRNA-HACE1),respectively.After cells were all cultured for 72 h under nor-moxic condition,cells were then cultured for 4 h in a hypoxic chamber,followed by reoxygenation for 2 h to simulate acute oxidative stress injury in H9C2 cardiomyocytes.Cell viability was assessed using CCK-8,the apoptosis rate in each group was measured via Hoechst 33342 staining,and HO-1 protein expression level as an indicator of oxidative stress in-jury was detected by Western blotting.Results The cell viability in the control group,HACE1-OE group,and shRNA-HACE1 group was 67.0%±4.4%,75.2%±4.2%,and 56.7%±3.0%,respectively,with statistically significant dif-ferences between these groups(all P<0.05).Apoptosis rates of the control group,HACE1-OE group,and shRNA-HACE1 group were 18.90%±1.02%,13.30%±0.86%,and 28.70%±0.67%,and there were statistically signifi-cant differences between these groups(all P<0.05).The relative expression levels of HO-1 protein in the control group,HACE1-OE group,and shRNA-HACE1 group were 1.00±0.17,0.71±0.19,and 1.96±0.28,and there were statistically significant differences between these groups(all P<0.05).Conclusion Transfection with HACE1 gene overexpression viral plasmid in H9C2 cardiomyocytes with acute oxidative stress injury significantly promotes cell viability,inhibits apoptosis,and reduces HO-1 protein expression;conversely,transfection with HACE1 gene knock-down viral plasmid leads to decreased cell viability accompanied by increased apoptosis rate and elevated HO-1 protein expression.
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