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claudin5的影响

摘要

M edicine o fM inistry o f Educa tion and Beijing, Dongzhim en Hospita l,Be ijing University o f Chinese M edicine(Be ijing 100700), China ABSTRACT :O bjective To observe the effects o f hypoxia on expression o f tight junction claudin 5 o f bra in m icrovascular en-do the lia l ce ls and the ro le o fQ ingka iling injection.M e thods M ouse cerebra lm icrovascula ture endo the lia l ce ls o f Ba lb/c w as cul-tured and divided into five g roups :Contro lg roup,m ode lg roup, Q ingka iling low, m edium and high concentra tion g roups. The m ode lo f ischem ic injury w as established by oxygen and sugar deprived fo r 24 h.Rea l T im e PCR w as used to eva lua te the gene expression o f claudin 5,and pro te in expression w as tested by w estern blo t.Results C om pared w ith contro l g roup,the expression o f claudin5 w as reduced significantly after oxygen and sugar be deprived fo r 24 h( P < 0.05). Com pared w ith the m ode l g roup, gene expres-sion in the low and high dose o fQ ingka iling injection increased significantly and claudin 5 pro te in im proved obviously in the m edium g roup.Conclusion Q ingka iling injection had cerebra lpro tection ro le in ischem ic stroke by recovering the barrier function through in-creasing the expression o f claudin 5.%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Ba lb/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 C laudin 5mRNA的转录水平，w estern blo t法检测蛋白表达。结果缺氧损伤后，C laudin 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。