目的 建立模拟体内血脑屏障（blood-brain barrier，BBB）模型，并运用该模型对丹酚酸组分中可能作用于中枢神经系统（CNS）的活性成分进行筛选，为CNS药物的快速高通量筛选提供实验依据。方法 将人脑微血管内皮细胞（HBMEC）和人星形胶质细胞（HA）进行共培养，模拟体内BBB，建立细胞培养水平的模型，实时考察模型形成过程中细胞的跨膜电阻值（TEER）及通透性。并运用该模型对丹酚酸组分中可能作用于CNS的有效活性成分进行筛选，利用液质联用技术对筛选出的化合物进行初步鉴定。结果 HA诱导HBMEC产生较高的跨膜电阻，于第10天达300 Ω/cm²，荧光素钠检测内皮细胞形成的紧密连接，其跨体外BBB模型的渗透系数为（2.659±0.730）×10-3 cm/min。利用该模型从丹酚酸组分中筛选出至少有6个成分能够穿越BBB模型，这6个成分被确认为丹参素、原儿茶酸、咖啡酸、异阿魏酸、紫草酸、迷迭香酸。结论 建立的体外模型在跨膜电阻和通透性方面具备了在体的基本特性，为CNS药物的快速、高通量筛选提供实验依据。

Objective To establish a blood-brain barrier (BBB) model for screening the effective ingredients of total salvianolic acids components probably effecting central nervous system (CNS), so as to provide basis for the rapid high throughput screening of CNS drugs. Methods The permeability parameters and transendothelial electrical resistance (TEER) values showed that the co-cultured human brain microvascular endothelial cells (HBMEC)/HA (human astrocytes) model was of integrity and vitality. The active components in total salvianolic acids were screened with this in vitro BBB model, and then analyzed by HPLC-MS. Results HA induced HBMEC to produce higher TEER in the first 10 d up to 300 Ω/cm², sodium fluorescein was used to detect the formation of tight junctions of endothelial cells, and the permeability coefficient of its cross-vitro BBB model was (2.659±0.730)×10-3 cm/min. We found that six compounds at least from total salvianolic acids could permeate across BBB model, in which danshensu, protocatechuic acid, caffeic acid, isoferulic acid, lithospermic acid, and rosmarinicacid were identified. Conclusion The co-cultured HBMEC/HA model combined with HPLC-MS analysis is a potential tool for screening of BBB permeable drug candidates from plant-derived materials.

国家自然科学基金资助项目（81573620，81473394）；苏州市医疗器械与新医药专项（ZXY2013022）