血脑屏障微流控芯片模型的构建及其中药活性成分跨膜转运研究  

Construction of a blood-brain barrier microfluidic chip model and evaluation of the permeability of active components in traditional Chinese medicine

作　　者：施艺玮 蔡颖 何晓莉 洪战英 柴逸峰[1] SHI Yi-wei;CAI Ying;HE Xiao-li;HONG Zhan-ying;CHAI Yi-feng(School of Pharmacy,Naval Medical University,Shanghai 200433,China;School of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]海军军医大学药学系,上海200433 [2]福建中医药大学药学院,福建福州350122

出　　处：《药学学报》2022年第3期802-808,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81872829,81673386,82173777)。

摘　　要：构建血脑屏障微流控芯片模型,应用于中药活性成分跨血脑屏障渗透性研究。该芯片模型由垂直交叉的两层通道及单层聚碳酸酯膜组成,采用原代人脑微血管内皮细胞,并能模拟血管剪切应力。通过活/死细胞染色和免疫荧光染色观察芯片上细胞生长状态,细胞生长状态良好,且在动态培养下细胞间黏附连接蛋白结构完整;考察该芯片模型对荧光示踪剂和3种模型药物的渗透性和P-糖蛋白(P-gp)的表达情况,结果显示荧光示踪剂和模型药物的渗透性与文献报道一致,芯片上P-gp表达和功能正常,表明该血脑屏障芯片模型具有完整的结构和功能。将构建成功的芯片模型应用于6种中药活性成分跨血脑屏障渗透性评价,采用HPLC-MS/MS法测定跨膜转运液中的药物浓度,获得各成分的P;结果。延胡索甲素P_(app)为(4.51±1.90)×10^(-7)cm·s^(-1)、延胡索乙素P_(app)为(9.10±6.59)×10^(-7)cm·s^(-1)、欧前胡素P_(app)为(9.38±2.53)×10^(-7)cm·s^(-1),而异欧前胡素、黄芩苷和绿原酸浓度低于定量限,推测其在芯片模型上渗透性较低。本研究成功构建血脑屏障微流控芯片模型,屏障功能更加完善且更接近生理环境,有望作为一种新的体外药物渗透性评价工具。A blood-brain barrier microfluidic chip platform for studying the permeability of active components in traditional Chinese medicine was developed. This model used primary human brain microvascular endothelial cells on a microfluidic chip consisting of two perpendicularly-crossing channels and a single layer porous polycarbonate membrane. The physiological shear stress in the human vasculature was also modeled in this device.Cell viability on the chip was monitored by cell staining and immunofluorescence staining. The cells spread well and the structure of an intercellular adhesion protein was satisfactory. The permeability of fluorescent tracers and three model drugs and the functional expression of P-glycoprotein(P-gp)on the blood-brain barrier were investigated. The results show that the apparent permeability coefficients(P;) of the fluorescent tracers and three model drugs were consistent with those reported in the literature, and P-gp on the chip showed normal function,indicating that there was a complete structure and a functional BBB. The permeability of six active components of traditional Chinese medicine was investigated through this microfluidic chip and the drug concentration was determined by HPLC-MS/MS to obtain the P_(app)of each component. The P;of corydaline was(4.51 ± 1.90)×10^(-7)cm·s^(-1), the P_(app)of tetrahydropalmatine was(9.10 ± 6.59) ×10^(-7)cm·s^(-1), and the P_(app)of imperatorin was(9.38 ± 2.53) ×10^(-7)cm·s^(-1);the concentration of isoimperatorin, baicalin and chlorogenic acid was below the limit of quantification, which suggested that isoimperatorin, baicalin and chlorogenic acid have poor permeability in this BBB chip. This blood-brain barrier microfluidic platform possesses a complete barrier function and near-physiological conditions and could be a valuable in vitro tool for drug permeability evaluation.

关 键 词：血脑屏障 微流控芯片 中药活性成分 药物渗透性 表观渗透系数 

分 类 号：R917[医药卫生—药物分析学]