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). Methods: CMECs were isolated from adult rat ventricles and exposed to simulated ischemia/reperfusion (SI/R). CMECs were randomly divided into three groups: control group, SI/R group, and SI/R+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. Results: Both cell viability and migration ability were impaired after SI/R (PO.05 vs control), and the apoptosis index increased as compared with that in the control group (PO.05). In addition, the SI/R group exhibited significantly higher celluar ROS level and higher expression of Drpl and Fisl protein than that in the control group (P<0.05). Administration of Mdivi-1 during reperfusion dramatically 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.
展开▼
