醒脑静注射液对大鼠脑出血模型血脑屏障通透性及相关蛋白的影响  

Effects of Xingnaojing injection on permeability of blood-brain barrier and related proteins in cerebral hemorrhage model rats

作　　者：王丽琴 孙逸坤[1] 王晓峰 李媛媛[1] 宋珂 刘浩琦 常静玲[1] 高永红[1] WANG Liqin;SUN Yikun;WANG Xiaofeng;LI Yuanyuan;SONG Ke;LIU Haoqi;CHANG Jingling;GAO Yonghong(Key Laboratory of Chinese Internal Medicine of Ministry of Education and Beijing Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700,China)

机构地区：[1]北京中医药大学东直门医院中医内科学教育部和北京市重点实验室,北京中医药大学脑病研究院,100700

出　　处：《环球中医药》2021年第6期993-999,共7页Global Traditional Chinese Medicine

基　　金：国家自然科学基金(81673899)。

摘　　要：目的观察醒脑静注射液对脑出血(intracerebral hemorrhage,ICH)大鼠血脑屏障通透性及其相关蛋白表达的影响。方法将27只雄性SD大鼠随机分为假手术组9只和模型组18只,采用右侧尾状核微量注射胶原酶Ⅶ的方法制备大鼠脑出血模型。将造模成功的大鼠随机分为模型组和醒脑静组,每组9只。连续给药3天后,采用伊文思蓝(evens blue,EB)法观察各组大鼠血脑屏障通透性;透射电镜观察脑组织的超微结构并分析计算星形胶质细胞足突的水肿面积;免疫组化法检测脑组织水通道蛋白4(aquaporin 4,AQP4)的表达;蛋白免疫印迹法(Western blot,WB)检测脑组织AQP4、血管内皮生长因子(vascular endothelial growth factor,VEGF)、闭合蛋白(Occludin)、紧密连接蛋白(tight junction protein,ZO-1)的表达。分别比较假手术组与模型组、模型组与醒脑静组血脑屏障的EB渗透程度、足突水肿面积以及血脑屏障相关蛋白的表达。结果(1)与假手术组相比,模型组大鼠脑出血后血脑屏障EB渗透性增加;与模型组相比,醒脑静组大鼠脑组织血脑屏障EB渗透性降低。(2)与假手术组相比,模型组大鼠脑组织中星形胶质细胞足突水肿面积明显增加,星形胶质细胞和血管内皮细胞结构明显被破坏,AQP4、VEGF蛋白表达升高(P<0.05);与模型组相比,醒脑静组大鼠脑出血后脑组织水肿减轻,星形胶质细胞以及血管内皮细胞的结构有所改善,AQP4、VEGF蛋白表达显著降低(P<0.05)。(3)与假手术组相比,模型组大鼠脑组织Occludin、ZO-1表达降低(P<0.05);与模型组相比,醒脑静组大鼠脑组织Occludin、ZO-1蛋白表达升高(P<0.05)。结论醒脑静注射液可通过减轻星形胶质细胞足突水肿,改善血脑屏障结构和相关蛋白表达,降低血脑屏障通透性可能是其减缓脑出血后脑组织损伤的作用机制。Objective To observe the effects of Xingnaojing injection on permeability of blood brain barrier(BBB)and related protein expression in rats with cerebral hemorrhage.Methods 27 male SD rats were randomly divided into sham operation group with 9 rats,and model group with 18 rats.The cerebral hemorrhage rat model was established by using the right caudate nucleus trace injection of collagenaseⅦ.The successful modeling rats were randomly divided into model group and Xingnaojing group,with 9 rats in each group.After 3 days of continuous administration,Evens blue(EB)method was used to observe the blood-brain barrier permeability of rats in each group.The ultrastructure of brain tissue was observed by transmission electron microscopy and the edema area of astrocyte foot process was analyzed and calculated.The expression level of aquaporin 4(AQP4)in brain tissue was detected by immunohistochemistry.The expression levels of AQP4,vascular endothelial growth factor(VEGF),occludin and tight junction protein(ZO-1)in brain tissue of rats were detected by Western blot.The degree of EB penetration,the area of foot process edema and the expression of BBB related proteins were compared among sham operation group,model group and Xingnaojing group.Results(1)Compared with sham operation group,the EB permeability of BBB after cerebral hemorrhage in model group increased.Compared with model group,the EB permeability of BBB in brain tissue of rats in Xingnaojing group.(2)Compared with sham operation group,the area of foot process edema of astrocytes in the brain tissue of rats in model group was significantly increased,the structure of astrocytes and vascular endothelial cells was significantly destroyed,and the protein expression levels of AQP4 and VEGF were increased(P<0.05).Compared with model group,after cerebral hemorrhage,the edema of brain tissue was alleviated in Xingnaojing group,the structure of astrocytes and vascular endothelial cells was improved,and the protein expression levels of AQP4 and VEGF were significantly decrea

关 键 词：脑出血 醒脑静注射液 血脑屏障 星形胶质细胞 内皮细胞 水通道蛋白4 紧密连接蛋白 

分 类 号：R285.5[医药卫生—中药学]