清开灵注射液对小鼠脑微血管内皮细胞缺氧模型紧密连接蛋白claudin-5的影响

目的 观察缺氧对脑微血管内皮细胞紧密连接蛋白claudin-5 表达的影响及清开灵的干预作用.方法 体外培养Balb/c 小鼠脑微血管内皮细胞,分为正常组、模型组和清开灵小中大剂量组,采用氧糖剥夺24 h 模拟缺血损伤,荧光定量PCR 法检测Claudin-5mRNA 的转录水平,western blot 法检测蛋白表达.结果 缺氧损伤后,Claudin 5mRNA 和蛋白表达水平明显降低（P 〈0.05）;与模型组比较,清开灵小、中、大各剂量组都有改善趋势,其中小、大剂量对基因转录水平提高明显,中剂量对蛋白表达的增强更显著,具有统计学意义（P 〈0.05）.结论 清开灵通过调节紧密连接蛋白claudin-5 表达,改善血脑屏障通透性从而阻抑脑缺血反应过程.

Effects of Qingkailing Injection on Expression of Tight Junction Claudin 5in the Hypoxia Model of Brain Microvascular Endothelial Cells

Objective To observe the effects of hypoxia on expression of tight junction claudin-5 of brain microvascular en- dothelial cells and the role of Qingkailing injection.Methods Mouse cerebral microvasculature endothelial cells of Balb/c was cul- tured and divided into five groups：Control group,model group,Qingkailing low,medium and high concentration groups.The model ofischemic injury was established by oxygen and sugar deprived for 24 h.Real Time PCR was used to evaluate the gene expressionof claudin 5,and protein expression was tested by western blot.Results Compared with control group,the expression of claudin5 was reduced significantly after oxygen and sugar be deprived for 24 h（P 〈0.05）.Compared with the model group,gene expres- sion in the low and high dose of Qingkailing injection increased significantly and claudin-5 protein improved obviously in the mediumgroup.Conclusion Qingkailing injection had cerebral protection role in ischemic stroke by recovering the barrier function through in- creasing the expression of claudin-5.