加味四君子汤对脑缺血大鼠脑组织Occludin,ZO-1,Claudin-1蛋白及其mRNA表达的影响

Effect of Modified Sijunzi Tang on Protein and mRNA Expressions of Occludin,ZO-1 and Claudin-1 in Cerebral Ischemia Rats

目的:探讨加味四君子汤对脑缺血大鼠脑组织闭合蛋白(Occludin),密封蛋白-1(Claudin-1),闭锁连接蛋白-1(ZO-1)表达的影响,并探讨相关的作用机制。方法:将60只雄性SD大鼠随机分为假手术组,模型组,依达拉奉组(3.2×10-3g·kg-1),加味四君子汤加减低、中、高剂量组(4.77,9.54,19.08 g·kg-1)。采用线栓法制备大脑中动脉栓塞(MCAO)模型,治疗7 d后处死,运用蛋白免疫印迹法(Western blot)检测大鼠缺血侧脑皮质区Occludin,ZO-1,Claudin-1蛋白的表达,运用实时荧光定量聚合酶链式反应(Real-time PCR)检测大鼠缺血侧脑皮质区Occludin,ZO-1,Claudin-1 mRNA表达。结果:与假手术组比较,模型组Occludin,Claudin-1,ZO-1蛋白显著减少(P<0.01);与模型组比较,依达拉奉组、加味四君子汤各剂量组Occludin,Claudin-1,ZO-1蛋白均显著增高(P<0.01)。与假手术组比较,模型组大鼠Occludin,Claudin-1,ZO-1 mRNA表达下调(P<0.01);与模型组比较,依达拉奉组、加味四君子汤各剂量组大鼠Occludin,Claudin-1,ZO-1 mRNA表达上升(P<0.01)。结论:加味四君子汤可能通过增加紧密连接蛋白Occludin,ZO-1,Claudin-1的表达,保护血脑屏障,减轻缺血性脑卒中大鼠脑水肿。

Objective:To investigate the effect of modified Sijunzi Tang on protein and mRNA expressions of Occludin,Claudin-1 and zonula occludens-1(ZO-1)in cerebral ischemia rats.Method:Totally 60 male Sprague-Dawley rats were randomly divided into sham operation group,model group,edaravone group,smalldose modified Sijunzi Tang group,middle-dose modified Sijunzi Tang group and high-dose modified Sijunzi Tang group.The middle cerebral artery occlusion(MCAO)model was prepared by suture method.After 7 days of treatment,the modeling group was put to death.Western blot was used to detect the contents of Occludin,Claudin-1 and ZO-1 in the ischemic cerebral cortex of rats.Detection of Occludin,ZO-1,Claudin-1 mRNA expression in the ischemic cortex of rats by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Result:Compared with the sham operation group,protein expressions of Occludin,Claudin-1,and ZO-1 in the model group were significantly down-regulated(P<0.01).Compared with the model group,protein expressions of Occludin,ZO-1,Claudin-1 in the positive control group and modified Sijunzi Tang groups increased significantly,with statistically significant differences(P<0.01).Compared with the Sham operation group,the expression of Occludin,Claudin-1,and ZO-1 mRNA in the model group was down-regulated(P<0.01).Compared with the model group,mRNA expressions of Occludin,Claudin-1,ZO-1 in the positive control group and modified Sijunzi Tang groups increased,with statistically significant differences(P<0.01).Conclusion:Modified Sijunzi Tang may protect the blood-brain barrier and reduce brain edema in ischemic stroke rats by increasing the expression of tight junction proteins Occludin,ZO-1 and Claudin-1.