Development of a humanized in vitro blood-brain barrier model to screen for brain penetration of antiepileptic drugs
Development of a humanized in vitro blood-brain barrier model to screen for brain penetration of antiepileptic drugs
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DOI：
10.1111/j.1528-1167.2006.00960.x
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发表时间：
2007-03-01
期刊：
影响因子：
5.6
通讯作者：
Janigro, Damir
中科院分区：
文献类型：
--
作者：
Cucullo, Luca;Hossain, Mohammed;Janigro, Damir
目的：研究药物穿过血脑屏障（BBB）的生物技术突破将是使用可重复的体外模型，该模型再现了BBB原位的功能，结构和病理特性。我们建立了一个人源化的动态体外血脑屏障模型（DIV-BBB），该模型基于人微血管内皮细胞（HBMEC）与人脑星形胶质细胞（HA）的共培养，HBMEC来源于“正常”和耐药的癫痫脑组织，HA来源于癫痫患者或对照组。HBMEC暴露于由管腔内流动产生的生理水平的剪切应力。在对照组和耐药DIV-BBB中测量了[H-3]蔗糖、[C-14]苯妥英和[C-14]地西泮的渗透性，并使用和未使用MDR 1抑制剂XR 9576进行预处理。通过跨内皮电阻测量（TEER）监测BBB完整性。通过测量葡萄糖消耗和乳酸production.Results：P-蔗糖和TEER值不依赖于所使用的内皮细胞的起源（癫痫或正常）的细胞生长和活力进行了评估。耐药血脑屏障模型中的对苯妥英钠浓度（1.54 x 10（-6）cm/s）比对照组（1.74 x 10（-5）cm/s）低10倍。XR 9576的MDR 1阻断仅在耐药培养物中有效（增加3.5倍）。对照和耐药DIV-BBB中的P-地西泮不受XR 9576的影响，并且不依赖于内皮细胞的癫痫或对照来源。癫痫胶质细胞的pharmacoresistance的整体贡献是微不足道的。结论：这些结果表明，对于所使用的物质，人源化的DIV-BBB概括的生理渗透性特性的血脑屏障在体内，也能够模仿耐药血脑屏障表型。
Purpose: A biotechnologic breakthrough for the study of drug permeability across the blood-brain barrier (BBB) would be the use of a reproducible in vitro model that recapitulates the functional, structural, and pathologic properties of the BBB in situ. We developed a humanized dynamic in vitro BBB model (DIV-BBB) based on cocultures of human microvascular endothelial cells (HBMECs) from "normal" and drug-resistant epileptic brain tissue with human brain astrocytes (HAs) from epilepsy patients or controls.Methods: HBMECs and HAs were cocultured for 28 days in polypropylene capillaries. HBMECs were exposed to physiologic levels of shear stress generated by intraluminal flow. Permeability to [H-3]sucrose, [C-14]phenytoin, and [C-14]diazepam was measured in control and drug-resistant DIV-BBB with and without pretreatment with the MDR1 inhibitor XR9576. BBB integrity was monitored by transendothelial electrical resistance measurements (TEERs). Cell growth and viability were assessed by measurement of glucose consumption and lactate production.Results: P-Sucrose and TEER values did not depend on the origin of the endothelium used (epileptic or normal). P-Phenytoin was 10-fold less (1.54 x 10(-6) cm/s) in drug-resistant BBB models than in controls (1.74 x 10(-5) cm/s). MDR1 blockade with XR9576 was effective (3.5-fold increase) only in drug-resistant cultures. P-Diazepam in control and drug-resistant DIV-BBB was not affected by XR9576 and did not depend on the epileptic or control origin of endothelia. The overall contribution of epileptic glia to pharmacoresistance was negligible.Conclusions: These results show that, for the substances used, the humanized DIV-BBB recapitulates the physiologic permeability properties of the BBB in vivo and is also capable of mimicking a drug-resistant BBB phenotype.