线粒体分裂抑制剂Mdivi-1对大鼠心肌微血管内皮细胞缺血再灌注损伤的保护作用

Protective Effects of Mdivi-1 against Cardiac Microvascular Endothelial Cells Injury Induced by Ischemia-Reperfusion

目的:探讨Mdivi-1对缺血再灌注损伤后大鼠心肌微血管内皮细胞(cardiac microvascular endothelial cells,CMECs)的保护作用。方法:分离培养大鼠CMECs,建立缺血再灌注(SI/R)模型,随机分为对照组、SI/R组、和SI/R+Mdivi-1组。采用MTT法检测细胞增殖能力;Transwell法检测细胞迁移能力;Annexin V-FITC/PI双标记流式细胞术检测CMECs凋亡率;western blot法检测各组细胞Drp1,Fis1水平;活性氧检测试剂盒测细胞ROS水平。结果:与对照组比较,SI/R组和SI/R+Mdivi-1组细胞增殖和迁移能力明显降低,凋亡明显增加,Drp1,Fis1表达增高,ROS水平明显升高,结果具有统计学意义(P<0.05);与SI/R组比较,SI/R+Mdivi-1组细胞增殖和迁移能力明显增加,凋亡明显降低,ROS水平明显降低,结果具有统计学意义(P<0.05),而Drp1,Fis1表达则无明显改变。结论:Mdivi-1可减轻CMECs的缺血再灌注损伤,这种保护作用可能是通过抗氧化应激来实现的。

Objective： To investigate the protective effects of Mdivi-1 against ischemia/reperfusion injury in cardiac microvascular endothelial cells （CMECs）. Mothods： CMECs were isolated from adult rat ventricles and exposed to simulated ischemia/reperfusion （SFR）. CMECs were randomly divided into three groups： control group, SFR group, and SFR＋Mdivi-1 group. Cell viability was measured by MTT assay and migration ability was detected by Transwell method. Flow cytometry based on Annexin V-FITC/PI double staining was used to detect apoptotic rates. The expression of Drpl and Fisl were analyzed by Western blot. Reactive oxygen species （ROS） levels were detected by a commercial reactive oxygen species assay kit. Rosults： Both cell viability and migration ability were impaired after SFR （P〈0.05 vs control）, and the apoptosis index increased as compared with that in the control group （P〈0.05）. In addition, the SFR group exhibited significantly higher celluar ROS level and higher expression of Drp 1 and Fisl protein than that in the controlgroup （P〈0.05）.AdrninistrationofMdivi-1 dufing reperfusion drarnatically attenuated the dysfunction of CMECs and decreased the level of ROS. However, the expression of Drpl and Fisl protein were not ultered. Conclusion： Mdivi-1 exerts protective effects against ischemia/reperfusion injury in CMECs, partly through inhibiting mitochondria fission and reducing celluar oxidative stress.