白藜芦醇苷对脑缺血再灌注损伤小鼠血脑屏障的保护作用研究  

Protective effect of polydatin on blood-brain barrier in mice after transient focal cerebral ischemia

作　　者：刘洋 李嘉民 陈荣 张祥建[3] LIU Yang;LI Jia-min;CHEN Rong;ZHANG Xiang-jian(Department of Neurology,Tianjin Huanhu Hospital,Tianjin300350,China;Department of Neurology,The First Hospital of Shijiazhuang City,Shijiazhuang 050011,China;Department of Neurology,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]天津市环湖医院神经内科,天津300350 [2]石家庄市人民医院神经内科,石家庄050011 [3]河北医科大学第二医院神经内科,石家庄050000

出　　处：《天津医科大学学报》2022年第3期278-283,共6页Journal of Tianjin Medical University

基　　金：河北省卫生健康委员会指令项目(20190061)。

摘　　要：目的:观察白藜芦醇苷(PD)对脑缺血再灌注损伤小鼠血脑屏障的保护作用及对紧密连接蛋白的影响。方法:CD-1小鼠(102只)应用线栓法制作短暂性大脑中动脉闭塞(tMCAO)模型。随机将实验动物分成4组:假手术组(28只)、缺血再灌注组(28只)、白藜芦醇苷低剂量组(30 mg/kg,18只),白藜芦醇苷高剂量组(60 mg/kg,28只)。于缺血再灌注后24 h进行行为学评分、TTC染色、干湿重测定、伊文思蓝染色,应用Western印迹、免疫荧光检测内皮细胞标志物血小板内皮细胞黏附分子-1(CD-31)、紧密连接蛋白闭锁小带蛋白-1(ZO-1)、咬合蛋白(Occludin)、闭合蛋白-5(Claudin-5)的表达情况。结果:(1)与缺血再灌注组相比,白藜芦醇苷高剂量组改善了缺血再灌注小鼠行为学评分、减少了脑梗死体积[(33.76%±13.27%)vs.(57.47%±9.53%),P<0.05]、减轻了缺血再灌注小鼠脑水肿程度[(75.62%±7.95%)vs.(85.72%±6.20%),P<0.05]、降低了伊文思蓝渗出量[(1.49±0.47)vs.(2.13±0.44),P<0.05]。(2)Western印迹结果显示白藜芦醇苷上调了ZO-1、Occludin、Claudin-5(均P<0.05)的表达水平。(3)免疫荧光结果显示:白藜芦醇苷组上调了CD-31在皮层缺血半暗带区的表达,且可见微血管形态相较缺血再灌注组更加光滑、完整,同时白藜芦醇苷减少了ZO-1、Claudin-5的破坏。结论:白藜芦醇苷的干预对缺血再灌注损伤小鼠起到了保护血脑屏障的作用,上调了紧密连接蛋白ZO-1、Occludin、Claudin-5的表达水平。Objective:To investigate the protective effect of polydatin(PD)on the blood-brain barrier and tight junction proteins in ischemia-reperfusion mice.Methods:A total of 102 CD-1 mice were subjected to transient middle cerebral artery occlusion model(tMCAO).The experimental animals were divided into 4 groups randomly:Sham group(n=28),tMCAO group(n=28),PD-L group(polydatin 30 mg/kg,n=18),PD-H group(polydatin 60 mg/kg,n=28).Neurological deficit scores,2,3,5-triphenyltetrazolium chloride(TTC),the standard wet-dry method,Evans blue exudation was evaluated 24 h after ischemia-reperfusion.Western blotting and immunofluorescence was applied to detect the expression level of endothelial cell markers platelet endothelial cell adhesion molecule-1(cd-31),tight junction protein atresia Band-1(ZO-1),occludin and claudin-5.Results:(1)Compared with the tMCAO group 24 h after tMCAO,the neurological deficit score was ameliorated,the infarct volume was reduced[(33.76%±13.27%)vs.(57.47%±9.53%),P<0.05],the brain water content was reduced[(75.62%±7.95%)vs.(85.72%±6.20%),P<0.05],the permeability of blood-brain barrier was decreased[(1.49±0.47)vs.(2.13±0.44),P<0.05]in PD-H group respectively.(2)Western blotting showed that the level of ZO-1,Occludin,Claudin-5,was up-regulated significantly in the PD group 24 h after operation,compared with tMCAO group(P<0.05,respectively).(3)Immunofluorescence showed that the expression of CD-31 was up-regulated in the cortex ischemic penumbra,and the tube shape of microvascular was smoother and more complete than that of the tMCAO group.Meanwhile,the disruption of ZO-1 and Claudin-5 staining was reduced in the PD group,compared with tMCAO group.Conclusion:Polydatin administration shows the protective effect of blood-brain barrier in mice with ischemia-reperfusion injury,meanwhile it can up-regulate the expression of ZO-1,Occludin,Claudin-5.

关 键 词：白藜芦醇苷 脑缺血再灌注损伤 血脑屏障 内皮细胞 紧密连接蛋白 

分 类 号：R743.3[医药卫生—神经病学与精神病学]