CYP2B6体外诱导活性评价模型的建立及其在中药活性成分筛选的应用  

Establishment of in vitro evaluation model for CYP2B6 induction and its application to screen inducers among TCMs

作　　者：徐聪[1] 徐思云[1] 胡海红[1] 余露山[1] 曾苏[1] 

机构地区：[1]浙江大学药学院药物分析与药物代谢研究室,浙江杭州310058

出　　处：《药学学报》2013年第1期119-124,共6页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81173120);国家重大专项资助项目(2012ZX09506001-004)

摘　　要：建立基于hPXR/CAR的CYP2B6药物诱导剂的体外高通量筛选模型,用于快速筛选通过hPXR/CAR途径对CYP2B6具有潜在诱导能力的中药活性成分,并帮助阐述其诱导机制。本论文采用双荧光素酶报告基因系统,即将插入有CYP2B6远端和近端启动子序列的报告基因质粒pGL3-CYP2B6-Luc与hPXR/CAR3表达质粒瞬时共转染HepG2细胞,用含有空白溶剂或药物的培养基培养48 h后裂解测定双荧光素酶活性。运用PXR和CAR的阳性诱导剂利福平和CITCO验证报告基因模型构建成功后,考察5种常见中药提取物和1种中药单体通过hPXR/CAR途径对CYP2B6的诱导作用。本研究构建的基于hCAR的CYP2B6报告基因模型中用hCAR3代替hCARwt是国内首次报道,并且配合基于hPXR的报告基因模型可以系统且有效地用于CYP2B6药物诱导剂的体外筛选并阐释其诱导机制。This paper is to report the development of a high-throughput in vitro system to screen hPXR/CAR mediated CYP2B6 drug inducers,and the application of it into the quick determination of induction activity toward CYP2B6 by various commonly used traditional Chinese medicines（TCMs） extract.Dual reporter gene assays were performed.The hPXR/CAR expression vectors and the reporter vector pGL3-CYP2B6-Luc involved in the distal and proximal promoters of CYP2B6 were co-transfected into HepG2 cells.Relative luciferase activities in cell lysate were analyzed after 48 h treatment of blank vehicle or drugs to determine the induction activity toward CYP2B6 by various commonly used TCMs extract.The positive hPXR/hCAR activators rifampicin and CITCO were applied to make sure that the reporter gene model was successfully established.Then 5 kinds of commonly used TCM extracts and 1 herbal compound were successfully investigated,some were found to activate hPXR or hCAR and therefore have the potential to induce CYP2B6 enzyme.This is the first domestic article to report the hCAR3-mediated CYP2B6 induction model and the establishment of a reporter gene system for hPXR/CAR-mediated CYP2B6 induction can be an effective and systemic in vitro method to investigate the drug inducers of CYP2B6 and to explain the mechanism involved.

关 键 词：CYP2B6 诱导 hPXR CAR 报告基因 中药 

分 类 号：R917[医药卫生—药物分析学]