Influence of ischemia reperfusion on viability and apoptosis of H9c2 myocardial cells and its mechanism

YANG Yingying; SUN Xiaoou; TAN Wen

doi:10.3969/j.issn.1002-266X.2016.39.005

Abstract：Objective To investigate the influence of ischemia reperfusion(IR)on cell viability and apoptosis of rat myocardial cells H9c2 as well as the potential molecular mechanism.Methods H9c2 cells were divided into groups A (control)and B (IR)and were seeded in 96-well plates by 1 ×104/well.After adhering for 24 h,the culture medium of group B was replaced by 100 μL hypoxia buffer and the cells were incubated in 94% N2 ,1% O2 ,and 5% CO2 for 90 min.Then we discarded the hypoxia buffer,and incubated them in normal environment with 100 μL/well DMEM of high glucose medium for another 90 min.The cell viability was detected by MTT,apoptosis by TUNEL,mitochondrial mem-brane potential (MMP)and reactive oxygen species (ROS)by laser scanning confocal microscope.Furthermore,heat shock protein 60 (HSP60),peroxiredoxin (Prx),thioredoxin (Trx)and mitochondria fission protein (Fis1 )were detec-ted by Western blotting.Results The cell viability of groups A and B was 0.77 ±0.01 and 0.40 ±0.02,respectively (P<0.05).The apoptosis rates in groups A and B were 2.24%±0.12%and 41.78%±1.43%(P<0.05).Intracellular ROS and mitochondrial membrane potential of group A was 0.58%±0.02% and 1.12%±0.05%,and was 1.13%± 0.05% and 0.76%±0.01%in the group B (all P<0.05).Compared with group A,the expression levels of HSP60 and Fis1 were increased,and the levels of Prx2 and Trx1 were decreased (all P<0.05).Conclusion After ischemia reper-fusion,the cell viability decreases and the apoptosis increases in H9c2 cells,and its mechanism may be that ischemia reperfusion promotes the expression of HSP60 and Fis1 protein and inhibits the expression of Prx2 and Trx1 protein.