远隔缺血后适应通过调控线粒体膜电位改善大鼠脑缺血/再灌注炎性损伤的机制研究

The mechanism of remote ischemic postconditioning to ameliorate the inflammatory injury of cerebral ischemia/reperfusion by regulating mitochondrial membrane potentials in rats

目的探究远隔缺血后适应介导线粒体膜电位调控脑缺血/再灌注损伤大鼠炎性损伤的作用机制。方法30只SD雄性大鼠随机分为假手术组、模型组及远隔缺血后适应组,每组10只,建立脑缺血/再灌注损伤大鼠模型,即刻进行3个循环远隔缺血后适应。采用Longa评分法评价各组大鼠神经功能障碍评分;HE染色观察各组大鼠脑组织病理学形态变化;TUNEL检测神经细胞凋亡指数;JC-1法测定线粒体膜电位;ELISA试剂盒测定大鼠缺血区脑组织IL-1β、IL-18表达;Western blot检测缺血区脑组织中NLRP3、Caspase-1、ASC及GSDMD蛋白表达。结果远隔缺血后适应干预后大鼠神经功能障碍评分明显降低,缺血侧脑组织损伤明显减轻,细胞凋亡指数明显降低,线粒体膜电位水平明显升高,缺血区脑组织IL-1β、IL-18水平、NLRP3、Caspase-1、ASC及GSDMD蛋白水平明显降低。结论远隔缺血后适应改善大鼠脑缺血/再灌注炎性损伤可能与调控线粒体膜电位有关。

Objective To explore the mechanism of mitochondrial membrane potential mediated by remote ischemic postconditioning to regulate inflammatory injury in rats with cerebral ischemia/reperfusion injury.Methods 30male SD rats were randomly divided into sham group,model group and remote ischemic postconditioning group,with 10 rats in each group.The rat model of cerebral ischemia/reperfusion injury was established,and 3 cycles of remote ischemic postconditioning were immediately performed.The neurological dysfunction scores were evaluated by Longa scoring.The pathological changes of brain tissue in each group was observed by HE staining.The apoptosis index of nerve cells was observed by TUNEL staining.Mitochondrial membrane potential was measured by JC-1 method.The expressions of IL-1βand IL-18 were detected by ELISA.The expressions of NLRP3,Caspase-1,ASC and GSDMD were measured by Western blot.Results After remote ischemic postconditioning intervention,the scores of neurological dysfunction were significantly reduced.Ischemic lateral brain tissue damage was significantly reduced.The apoptosis index was significantly decreased.Mitochondrial membrane potential significantly was increased.The levels of IL-1β,IL-18,NLRP3,Caspase-1,ASC and GSDMD was significantly decreased.Conclusion The amelioration of inflammatory injury of cerebral ischemia/reperfusion in rats by remote ischemic postconditioning may be related to the regulation of mitochondrial membrane potential.