复氧前后抑制线粒体通透性转换对心肌细胞低氧/复氧损伤的影响

Effect of inhibiting mitochondrial permeability transition before/after reoxygenation on cardiomyocytes against hypoxia-reoxygenation injury.

目的观察复氧前后抑制线粒体通透性转换(MPT)对心肌细胞低氧/复氧损伤的影响。方法新生SD大鼠心肌细胞原代培养48h后按孔随机分为空白对照组、低氧/复氧组、复氧前处理组和复氧后处理组,分别在复氧前、后10min给予1μmol/L环胞菌素A,复氧50min后用激光共聚焦显微镜观察MPT,复氧120min后采用流式细胞术和western blotting法分别观察心肌细胞凋亡和线粒体Cyt C释放。结果与低氧/复氧组比较,复氧前处理组细胞MPT明显减弱,细胞凋亡程度明显减轻,线粒体Cyt C释放显著减少。而复氧后处理组与低氧/复氧组比较差异无统计学意义。结论复氧前抑制线粒体通透性转换可以减轻心肌细胞低氧/复氧损伤,复氧后抑制线粒体通透性转换不能产生细胞保护作用。

Objective To explore the influence of inhibiting mitochondrial permeability transition before/after reoxyganation on cardiomyocytes against hypoxia-reoxygenation injury.Methods The primary cultured neonate SD rat cardiomyocytes were randomly divided into Control,H/R,CsA 1 and CsA 2 which were treated with 1 μmol/L CsA plus H/R at 10 minutes before/after reoxygenation.At 50 min after reoxygenation,the MPT was monitored by confocal imaging.At 120 min after reoxygenation,the apoptosis index were detected with Flow cytometry and the releases of cytochrome C from mitochondria were detected through western-blotting.Results Compraed to H/R group,MPT in CsA 1 group was inhibited obviously;The apoptosis and releases of cytochrome C in CsA 1 group were lessened obviously.There is no statistical difference between CsA 2 and H/R group.Conclusion Inhibiting MPT before reoxygenation can protect cardiomyocytes against hypoxia-reoxygenation injury.Inhibiting MPT after reoxygenation has no protective effect.