补阳还五汤对大鼠脑微血管内皮细胞氧糖剥夺再灌注损伤的保护作用  

Buyang Huanwu Tang Protects Rat Brain Microvascular Endothelial Cells Injured by Oxygen-glucose Deprivation-reperfusion

作　　者：范文涛[1] 王利胜[1] 范有明[2] 李周[1] 廖卫国[1] 

机构地区：[1]广州中医药大学中药学院,广州510006 [2]第三军医大学高原军事医学系,重庆400038

出　　处：《中国实验方剂学杂志》2016年第14期111-115,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81373973);广东省省级科技计划项目(2013B032500015)

摘　　要：目的:建立体外血脑屏障模型,研究补阳还五汤对氧糖剥夺再灌注损伤状态下血脑屏障功能及紧密连接相关蛋白表达的影响。方法:分离培养原代大鼠脑微血管内皮细胞,建立体外血脑屏障氧糖剥夺再灌注损伤模型,将细胞分为空白组、模型组、补阳还五汤提取液处理组,采用细胞计数试剂盒(CCK-8)法检测细胞活性;采用转移小室(Transwell)检测牛血清白蛋白(BSA)的透过量;蛋白质印迹(Western blot)实验检测紧密连接蛋白闭合蛋白(Occludin)和紧密连接相关蛋白-1(ZO-1)的表达;利用实时荧光定量聚合酶链式反应(q PCR)检测紧密连接蛋白Occludin,ZO-1 mRNA的表达。结果:与空白组比较,模型组细胞活力降低,BSA的透过率明显升高,Occludin,ZO-1蛋白及其mRNA的表达量明显降低(P<0.05);与模型组比较,补阳还五汤预处理后细胞存活性明显升高,BSA的透过率降低,Occludin,ZO-1蛋白及其mRNA的表达量明显升高(P<0.05)。结论:补阳还五汤能够降低氧糖剥夺再灌注处理对体外血脑屏障模型的损伤,上调紧密连接蛋白ZO-1,Occludin蛋白及其mRNA的表达量。Objective： To establish an in vitro blood-brain barrier, in order to study the effects of Buyang Huanwu Tang on blood-brain barrier function and associated protein expression under the condition of oxygen and glucose deprivation-reperfusion injury. Method： The in vitro oxygen and glucose deprivation- reperfusion injury model was established by culturing the isolated primary rat brain microvascular endothelial cells. The cells were divided into three groups, namely blank group, model group and Buyang Huanwu Tang group, and then were processed by following steps. Cell counting kit （CCK-8） method was used to detect cell activity; Transwell assay was used to detect the permeation of bovine serum albumin （BSA） ; Western blot assay was used to detect the protein expressions of Occludin, ZO-1. Real-time fluorescent quantitative PCR （qPCR） was used to detect the mRNA expressions of Occludin and ZO-1. Result： Compared with the blank group, the model group showed significant decreases in cell activity, Occludin and ZO-1 protein and their mRNA expressions, and increase in BSA permeation rate （P 〈 0.05 ）. In Buyang Huanwu Tang group, cell survival rate was significantly increased; bovine serum albumin （BSA） transmittance reduced; Occludin, ZO-1 protein and mRNA expression increased （P 〈 0.05 ）. Conclusion ： Buyang Huanwu Tang can reduce the oxygen glucose deprivation-reperfusion damage in the blood-brain barrier model in vitro and up-regulate ZO-1, Occludin protein and mRNA expression.

关 键 词：补阳还五汤 脑微血管内皮细胞 氧糖剥夺再灌注 血脑屏障 紧密连接蛋白 

分 类 号：R285.5[医药卫生—中药学]