目的 建立体外模拟体内肠道细胞——人结肠癌上皮细胞（the human colon adenocarcinoma cell line，Caco-2）的Transwell模型，以此研究钩藤碱在Caco-2细胞模型上的跨膜转运特征。方法 采用聚酯碳酸酯膜连续培养Caco-2细胞21 d，形成致密的单层细胞模型。然后对影响钩藤碱在Caco-2细胞模型上转运特征的因素(包括浓度、时间及跨膜转运蛋白P-糖蛋白)进行考察；同时采用液相色谱-质谱法对溶液中钩藤碱的含量进行测定。结果 钩藤碱在Millicell系统上的转运量具有一定的浓度-时间依赖性，主要以主动转运的方式进行吸收，且随着时间和药物浓度的增加，转运量明显增加。不同浓度钩藤碱从细胞基底侧（basolateral, BL）到细胞刷状缘侧（apical，AP）的表观渗透系数（apparent permeability coefficient, Papp）与AP侧到BL侧的Papp比值均在1.5以上，且钩藤碱从BL侧到AP侧的外排速率明显高于从AP侧到BL侧的吸收速率。当加入P-糖蛋白抑制剂后，药物由BL侧向AP侧的转运量显著减少，而由AP侧到BL侧的转运量显著增加。结论 钩藤碱在Caco-2细胞上转运存在一定的浓度及时间依赖性，且P-糖蛋白介导钩藤碱在Caco-2细胞上转运。
Objective To establish an in vitro Transwell model of intestinal Caco-2 cells, and to investigate the transmembrane transport characteristics of rhynchophylline in the Caco-2 cell model. Methods Caco-2 cells were cultured with polycarbonate membrane for 21 consecutive days to form a dense monolayer cell model, and then the factors affecting the transport characteristics of rhynchophylline in Caco-2 cell model were assessed, including concentration, time, and the transmembrane transporter P-glycoprotein. Liquid chromatography-mass spectrometry was used to measure the content of rhynchophylline in solution. Results The amount of transport of rhynchophylline in the Millicell system showed a concentration-time-dependent manner to a certain degree, and rhynchophylline was mainly absorbed by active transport. The amount of transport increased significantly with the increase in time and drug concentration. Different concentrations of rhynchophylline had an apparent permeability coefficient of above 1.5 from basolateral（BL） side to apical（AP） side and from AP side to BL side, and the efflux rate of rhynchophylline from BL side to AP side was significantly higher than the absorption rate of rhynchophylline from AP side to BL side. After a P-glycoprotein inhibitor was added, there was a significant reduction in the amount of transport of rhynchophylline from BL side to AP side and a significant increase in the amount of transport of rhynchophylline from AP side to BL side. Conclusion The transport of rhynchophylline in Caco-2 cells shows a concentration-time-dependent manner, and P-glycoprotein mediates the transport of rhynchophylline in Caco-2 cells.
孟 杰,苏 瑞,廖 赟,袁 波,李 玲.钩藤碱在Caco-2细胞模型中吸收规律研究[J].安徽中医药大学学报,2020,39(5):60-65复制