Caspase Inhibitor Reduces Myocyte Cell Apoptosis Induced by Hypoxia in vitro

Objective To investigate the significance of Caspase-3 inhibitor in neonatal cardiomyocytes apoptosis induced by hypoxia.Methods The cultured Cardiomyocytes from 1～2 d newborn SD rats were randomly divided into 3 groups: control groups,ischaemic 72 h group and Caspase3 inhibitor group [addition of Z-DEVD-fmk(0.1 μmol/L)72 h before ischaemic treatment].Caspase-3,PARP mRNA and protein were detected by RT-PCR and Western blot analysis,the Caspase-3 activity was detected by caspase-3 fluorescent assay kit.Cardiomyocytes apoptosis was detected by flow cytometry.Results The flow cytometry analysis showed that the percentage of apoptosis cells in Caspase-3 inhibitor group was(14.93±2.47)%,and compared with the 72 h ischaemic group(48.43±4.18)% was significantly lower(P0.01).RT-PCR and western blot analysis showed that Caspase-3 mRNA,PARP and Caspase-3 protein expression in Caspase-3 inhibitor group were significantlylower than those in the 72 h ischeamic group(P0.01).Caspase-3 activity analysis showed that caspase-3 activity in caspase-3 inhibitor group reduced by 52.85% relative to the 72 h ischaemic group.Conclusions Caspase-3 inhibitor can efficiently inhibit apoptosis of neonatal cardiomyocytes after hypoxia insult.Caspase-3 inhibitor can efficiently inhibit expression of Caspase-3 protein and mRNA in cultured neonatal cardiomyocytes after hypoxia insult.Caspase-3 inhibitor can efficiently inhibit activity of Caspase-3 in cultured neonatal cardiomyocytes after hypoxia insult.