白芷香豆素成分对葛根素跨血脑屏障体内外转运的影响  

Effect of Angelica dahurica coumarins on the transport behavior of puerarin across blood-brain barrier in vitro and in vivo

作　　者：拓文静 宋继红 韩成坤 王建香 杨文雪 卢闻 TA Wen-jing;SONG Ji-hong;HAN Cheng-kun;WANG Jian-xiang;YANG Wen-xue;LU Wen(School of Pharmacy,Health Science Center,Xi'an Jiaotong University,Xi'an 710061,China)

机构地区：[1]西安交通大学医学部药学院,陕西西安710061

出　　处：《药学学报》2023年第5期1156-1164,共9页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(82173775)。

摘　　要：构建原代大鼠脑微血管内皮细胞(rat brain microvascular endothelial cells, BMEC)和大鼠星状胶质细胞(rat astrocytes, AS)共培养模型以研究葛根素-白芷香豆素成分联用后对葛根素在大鼠体外血脑屏障(blood-brain barrier, BBB)模型中转运及大鼠脑组织分布的影响。通过测定模型跨内皮电阻值、苯酚红通透性、BBB相关蛋白表达评价模型屏障功能。通透性实验和Western blot法分别研究了白芷香豆素成分对BMEC、大鼠脑组织通透性及BBB相关蛋白表达的影响。本研究中动物实验方案均获得西安交通大学医学部生物医学伦理委员会的批准(动物伦理号:2021-1329)。结果表明,构建的BMEC/AS共培养模型可用于药物体外跨BBB研究;与白芷香豆素成分联用后,葛根素在体外共培养模型及大鼠脑组织上的转运量显著提升;白芷香豆素成分可以增加BBB通透性,抑制P糖蛋白(P-glycoprotein, P-gp)、紧密连接重要蛋白闭锁连接蛋白-1 (zonula occludens-1, ZO-1)和闭合蛋白(occludin)的表达。白芷香豆素成分可能通过抑制P-gp及紧密连接蛋白表达而增加葛根素在脑组织中的含量。A BBB co-culture cell model consisting of rat brain microvascular endothelial cells(BMEC)and astrocytes(AS)was established to study the effect of Angelica dahurica coumarins on the transport behavior of puerarin across blood-brain barrier(BBB)in vitro and in vivo.The barrier function of this model was evaluated by measuring the transendothelial resistance,phenol red permeability and BBB related protein expression.The permeability assay and western blot methods were performed to study the effects of Angelica dahurica coumarins on the BBB permeability and the expression of BBB related protein.The animal experiment protocols in this study were approved by the Animal Ethics Committee of Xi'an Jiaotong University(Animal Ethics No.:2021-1329).The results showed that the established BMEC/AS co-culture model could be used to evaluate drug transport across BBB in vitro.After combined with Angelica dahurica coumarins,the transport capacity of puerarin was significantly increased in vitro and in vivo.Additionally,Angelica dahurica coumarins enhanced BBB permeability and inhibited the protein expression of P-glycoprotein(P-gp),zonula occludens-1(ZO-1)and occludin.Angelica dahurica coumarins might increase BBB permeability by inhibiting the expression of P-gp and tight junction protein,thereby increasing the content of puerarin in brain tissue.

关 键 词：葛根素 白芷香豆素成分 血脑屏障 脑转运 共培养细胞模型 

分 类 号：R917[医药卫生—药物分析学]