Effects of up regulation of GDF-15 expression on biological characteristics and PI3K/AKT signaling pathway in H9C2 cardiomyocytes induced by H2O2 / 中国免疫学杂志

ABSTRACT

Objective:To investigate the effect of up regulation of GDF-15 expression on the proliferation,apoptosis and PI3K/AKT signaling pathway of H9C2 cardiomyocytes induced by H2O2.Methods:CCK8 method was used to detect the proliferation of H9C2 cardiomyocytes treated with different concentrations of H2O2;H9C2 cells were divided into Control group,NC group,H2O2group,GDF-15+H2O2group,the cells were treated for 24 h,mRNA and protein expression of GDF-15 in each group were detected by RT-PCR and Western blot;the proliferation and apoptosis of the cells were detected by CCK8 and flow cytometry respectively;DCFH-DA probe was used to detect the level of ROS;the expression of Ki67,Bcl-2,Bax,PI3K and p-AKT protein was detected by Western blot.H9C2 cells were treated with 10 μmol/L LY294002(a PI3K/AKT signal pathway inhibitor),cell viability and apoptosis rate were detected by CCK8 assay and flow cytometryin espectively.Ki67,Bcl-2,Bax,PI3K and p-AKT protein expression were detected by Western blot.Results:Cell viability was inhibited after different concentrations H2O2treated H9C2 myocardial cells,which was concentration de-pendent (P<0.05),due to H9C2 cardiomyocytes treated with 200 μmol/L H2O2inhibited nearly half of cell proliferation,and were chosen as subjects.Compared with control group,mRNA and protein expression of GDF-15 in H2O2group were significantly increased, cell proliferation was decreased significantly,the apoptosis rate was increased,ROS level was increased,the expression of Ki67,Bcl-2, PI3K,p-AKT protein were decreased,Bax protein expression was increased(P<0.05).Compared with H2O2group,mRNA and protein expression of GDF-15 in GDF-15+H2O2group were significantly increased,cell proliferation was significantly increased,the apoptosis rate was decreased,ROS level was decreased,the expression of Ki67,Bcl-2,PI3K,p-AKT protein were increased,and Bax protein expression was decreased (P<0.05).The cell viability and protein expression of Bcl-2,PI3K and p-AKT in PI3K/AKT inhibitor group were significantly lower than those in GDF-15+H2O2group,and the apoptosis rate and Bax protein expression were significantly higher than those in GDF-15+H2O2group(P<0.05).Conclusion:Up regulation of GDF-15 expression promote the proliferation of H9C2 car-diomyocytes induced by H2O2and reduce apoptosis,and the mechanism may be related to the regulation of ROS levels,Ki67,Bcl-2,Bax expression and PI3K/AKT signaling pathway in cells.