山东医药2016，Vol.56Issue(39)：16-19,4.DOI:10.3969/j.issn.1002-266X.2016.39.005
缺血再灌注对大鼠心肌H9 c2细胞活性、凋亡的影响及其机制
Influence of ischemia reperfusion on viability and apoptosis of H9c2 myocardial cells and its mechanism
摘要
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.关键词
缺血再灌注/H9 c2心肌细胞/细胞活性/细胞凋亡/活性氧簇/线粒体膜电位/热休克蛋白60/过氧化物氧化还原酶/硫氧还原蛋白/线粒体分裂蛋白Key words
ischemia reperfusion/H9c2 cardiomyocytes/cell viability/apoptosis/reactive oxygen species/mito-chondrial membrane potential/heat shock protein 60/peroxiredoxin/thioredoxin/mitochondrial fission protein分类
医药卫生引用本文复制引用
杨莹莹,孙晓鸥,谭文..缺血再灌注对大鼠心肌H9 c2细胞活性、凋亡的影响及其机制[J].山东医药,2016,56(39):16-19,4.基金项目
华南理工大学自然科学基金资助项目（D2154630）。 （）
