健脾益气利水方对脑缺血大鼠脑水肿及其脑组织Occludin、Claudin-1、ZO-1蛋白表达的影响

Effects of Jianpi Yiqi Lishui Fomula on cerebral edema and the expression of Occludin, Claudin-1 and ZO-1 protein in brain of rats with cerebral ischemia

目的:探讨健脾益气利水方(JPF)对脑缺血大鼠脑水肿及脑组织紧密连接蛋白闭合蛋白(Occludin)、密封蛋白-1(Claudin-1)、闭锁连接蛋白-1(ZO-1)表达的影响。方法:将105只雄性SD大鼠随机分为假手术组,模型组,依达拉奉组,JPF小、中、大剂量组。采用线栓法制备大脑中动脉栓塞(MCAO)模型,治疗7d后处死,运用伊文思蓝(EB)渗出法检测血脑屏障的通透性,运用免疫组化法检测大鼠缺血侧脑组织Occludin、Claudin-1、ZO-1蛋白表达。结果:与假手术组比较,模型组缺血侧脑组织EB的含量显著增加(P<0.01),Occludin、Claudin-1、ZO-1阳性细胞表达显著降低(P<0.01);与模型组比较,依达拉奉组和JPF中、大剂量组大鼠缺血侧脑组织EB的含量明显降低(P<0.05),各给药组Occludin、Claudin-1、ZO-1阳性细胞表达明显增强(P<0.01,P<0.05)。结论:健脾益气利水法组方可能通过上调紧密连接蛋白Occludin、Claudin-1、ZO-1表达保护血脑屏障,减轻脑缺血再灌注大鼠脑水肿。

Objective: To study the effects of Jianpi Yiqi Lishui Fomula(JPF) on brain edema and the expression of tight junction protein Occludin, Claudin-1 and ZO-1 in brain of rats with cerebral ischemia. Methods: A total of 105 male SpragueDawley rats were randomly divided into sham-operation group, model group, Edaravone group, JPF low dose group, JPF medium dose group and JPF high dose group. The MCAO model was made by suture method. After 7 days of treatment, the rats were killed. The permeability of blood-brain barrier was measured by evans blue exudation method, and the expression of Occludin, Claudin-1 and ZO-1 protein in ischemic brain tissue of rats was detected by immunohistochemical method. Results: Compared with sham-operation group, the content of EB in ischemic brain tissue increased significantly in model group(P<0.01), and the expression of Occludin, Claudin-1 and ZO-1 in ischemic brain tissue decreased significantly(P<0.01). Compared with model group, the content of EB decreased significantly in the Edaravone group, JPF medium dose group and JPF high dose group(P<0.05), the expression of Occludin, Claudin-1 and ZO-1 increased significantly(P<0.01, P<0.05) in the ischemic brain tissue of rats in each JPF group. Conclusion: Jianpi Yiqi Lishui therapy can protect the blood-brain barrier by increasing the expression of tight junction protein such as Occludin, Claudin-1 and ZO-1, and alleviate cerebral edema in rats with cerebral ischemiareperfusion injury.