非接触式共培养体外血脑屏障模型的跨膜电阻及通透性  

Transendothelial electric resistance and permeability of in vitro model of no-contact co-culture blood-brain barrier

作　　者：张水华[1,2] 季龙凤 马璟 

机构地区：[1]福建医科大学福建省新药安全性评价中心,福建福州350108 [2]国家上海新药安全性评价研究中心,上海201203

出　　处：《中国药理学与毒理学杂志》2012年第6期882-887,共6页Chinese Journal of Pharmacology and Toxicology

基　　金：国家"重大新药创制"科技重大专项(2008ZX09305-006);福建医科大学博士科研启动基金(2011BS002);福建省政府专项基金(闽发改高技2009-648)~~

摘　　要：目的建立大鼠脑毛细血管内皮细胞(BCEC)与星形胶质细胞共培养血脑屏障模型并评价其功能。方法采用SD大鼠原代分离培养获得BCEC和星形胶质细胞。经细胞形态学观察、免疫组化检测相关抗原后建立非接触式共培养血脑屏障模型,测定共培养模型所形成的跨细胞电阻及荧光素钠的通透性。采用LC-MS检测6个化合物透过血脑屏障模型的通透性,并与文献报道的体内数据进行比较。结果培养的BCEC多数为短梭形外观,免疫组化检测可见细胞高表达因子Ⅷ;星形胶质细胞呈现具有细胞突起的典型形态,免疫组化检测可见细胞高表达胶质纤维酸性蛋白。共培养的体外血脑屏障模型跨BCEC单层的电阻值为(373±41)Ω·cm2,荧光素钠跨BCEC单层的通透性为(0.34±0.14)×10-3cm·min-1,符合体外血脑屏障模型要求。对所选6个化合物体内外透过BBB模型渗透系数的比较,表明具有一定的相关性(R2=0.7679,P<0.05)。结论建立的体外BBB模型在跨内皮电阻和通透性方面具备了在体BBB的基本特性,可以用于模拟体内环境,进行药物早期筛选方面的研究。OBJECTIVE To establish and evaluate a co-cukure model of blood-brain barrier （BBB） using primary rat brain capillary endothelial cells （BCEC） and astrocytes. METHODS Primary cultures of BCEC and astrocytes were prepared from SD rats. Two kinds of cells were identi- fied through cell morphology and immunohistochemistry. No-contact co-culture BBB model in vitro was developed and transendotheliar resistance and permeability of fluorescein sodium were meas- ured. Additionally in vitro BBB permeabilities of six compounds were measured by LC-MS and com- pared their results with in vivo from literatures. RESULTS The cultured BCEC possessed the spin- dle-shaped morphology and expressed factor VⅢ antigen. The cultured astrocytes possessed cell process morphology and expressed GFAP antigen. Transendothelial electric resistance （TEER） and permeability of fluorescein sodium were （ 373 ± 41 ） Ω· cm2 and （ 0.34 ± 0.14 ） × 10 -3 cm· min -1, respectively. These values were consistent with literatures. The correlation （R2 = 0. 7679, P 〈 0.05） between in vitro permeability of selected six compounds and the in vivo published data was calculated. CONCLUSION TEER and permeability of in vitro BBB model have similar properties as that of in vivo BBB model. This model can simulate environment in vivo BBB and be used for drug screening study.

关 键 词：毛细血管内皮细胞 脑 星形胶质细胞 血脑屏障 

分 类 号：R96[医药卫生—药理学]