以方剂配伍减毒为切入点，构建3D人体血脑屏障（BBB）模型，研究配伍对中药毒性成分脑部转运的影响。课题组前期与新加坡国立大学合作，采用光刻微加工技术制造出水凝胶微孔阵列芯片，并成功培养了多种细胞的3D体外模型。本课题拟进一步构建可模拟体内微环境的3D人体BBB模型，探索不同培养因素对BBB紧密连接和转运蛋白调控的影响；结合转运蛋白过表达细胞和动物模型，以具有中枢毒性的中药麻黄和附子为研究示范，从分子、细胞、3D微组织以及生物体水平阐明有毒生物碱成分的BBB转运机制；运用该模型诠释配伍对这些生物碱成分脑部摄取的拮抗作用，揭示配伍减毒机理，丰富祖国医学治疗思想，拓展中药药代动力学研发思路。

A 3D human blood-brain barrier (BBB) model using microwell arrays will be built, to study the central toxic attenuation by compatibility of Traditional Chinese Medicine. Working with collaborators from the National University of Singapore, we have successfully fabricated hydrogel microwell arrays by a photolithography to establish various in vitro 3D cell culture models. In this proposal, an in vitro 3D human BBB model will be developed, and the effects of culture conditions on the BBB tight junction and the regulation of drug transporters will be investigated. Combining with the transporter over-expressing cells and animals, Ephedra and Aconiti Lateralis Radix Praeparata will be used as the model with CNS toxicity to study the BBB transport mechanism of their toxic alkaloids. Furthermore, the effects on the BBB transport of these alkaloids by different compatibility will be investigated to explain the toxic attenuation mechanism. The present work will be of great help for the pharmacokinetic studies of the Traditional Chinese Medicine.