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目的研究脑缺血后血脑屏障通透性和紧密连接相关蛋白occludin和claudin-5的变化。方法线栓法制备大鼠大脑中动脉缺血模型,采用伊文氏兰(EB)法检测缺血后血脑屏障通透性的变化。应用免疫组织化学的方法观察occludin和claudin-5在缺血脑组织中的分布,采用RT-PCR、Wesrern blot法检测大鼠缺血脑组织occludin和claudin-5的mRNA和蛋白的表达变化。结果脑缺血2h后血脑屏障通透性显著增加(P<0.01),紧密连接相关蛋白occludin和claudin-5在脑微血管内皮细胞上呈阳性表达,occludin和claudin-5的mRNA和蛋白均比正常对照组显著降低(P<0.01)。结论脑缺血时血脑屏障通透性的增加可能与紧密连接相关蛋白occludin和claudin-5的降低相关。
Objective To study the changes of blood-brain barrier permeability and tight junction-associated proteins occludin and claudin-5 after cerebral ischemia. Methods The rat middle cerebral artery occlusion (MCAO) model was established by thread occlusion. The permeability of blood-brain barrier (BBB) after ischemia was measured by Evans blue (EB) method. The distribution of occludin and claudin-5 in ischemic brain tissue was observed by immunohistochemistry. The mRNA and protein expression of occludin and claudin-5 in ischemic brain tissue were detected by RT-PCR and Wesrern blot. Results The permeability of blood-brain barrier increased significantly (P <0.01) 2h after cerebral ischemia, and occludin and claudin-5 were positively expressed on brain microvascular endothelial cells. The mRNA and protein expressions of occludin and claudin-5 The normal control group was significantly lower (P <0.01). Conclusions The increased permeability of the blood-brain barrier during cerebral ischemia may be related to the decrease of tight junction-associated proteins occludin and claudin-5.