Angiopoietin-1 prevents rat cardiomyocytes from H_2O_2-induced apoptosis by regulating the activity of PI3K/Akt

Objective To investigated whether angiopoietin-1(Ang-1) has direct cytoprotective effects on rat cardiomyocytes against oxidative stress and its possible signaling pathway involved.Methods A model for primary culture of cardiomyocytes injuried by H2O2 was established,the cultured cardiomyocytes were divided randomly into four groups.The morphology of cardiomyocytes was observed by Hoechst-33342.Apoptosis of cardiomyocytes was determined by flow cytometry(FCM).The expression of p-AKT and active-caspase-3 was measured by Western blot analysis.Results The percentage of apoptosis cells in control group and Ang-1 intervention group were 2.13%±0.61%,31.20%±2.01%,compared with the induced group(48.16%±1.37%)was significantly lower(P0.01),andapoptosis rate of cointervention group was 47.42%±2.02%,has no significant difference as compared with that of induced group.Western blot analysis showed Ang-1 restored the phosphorylation of Akt and inhibited the activation of caspase-3,and these effects were reversed by the presence of LY294002.Conclusion Ang-1 has direct cytoprotective effect on cardiomyocytes against oxidative stress and this effect is mediated through PI3K/Akt pathway and inhibition of caspase-3 cleavage.