Xu Cong, Xu Si-Yun, Hu Hai-Hong, Yu Lu-Shan, Zeng Su
Department of Pharmaceutical Analysis and Drug Metabolism, College of Pharmaceutical Sciences, Zhejiang University, Hangzhou 310058, China.
Yao Xue Xue Bao. 2013 Jan;48(1):119-24.
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.
本文旨在报道一种用于筛选人孕烷X受体（hPXR）/组成型雄烷受体（CAR）介导的CYP2B6药物诱导剂的高通量体外系统的开发，以及将其应用于快速测定各种常用中药提取物对CYP2B6的诱导活性。进行了双报告基因测定。将参与CYP2B6远端和近端启动子的hPXR/CAR表达载体和报告载体pGL3-CYP2B6-Luc共转染到HepG2细胞中。在用空白载体或药物处理48小时后，分析细胞裂解物中的相对荧光素酶活性，以确定各种常用中药提取物对CYP2B6的诱导活性。应用阳性hPXR/hCAR激活剂利福平和CITCO以确保报告基因模型成功建立。然后成功研究了5种常用中药提取物和1种草药化合物，发现其中一些可激活hPXR或hCAR，因此具有诱导CYP2B6酶的潜力。这是国内首篇报道hCAR3介导的CYP2B6诱导模型的文章，建立用于hPXR/CAR介导的CYP2B6诱导的报告基因系统可以成为一种有效且系统的体外方法，用于研究CYP2B6的药物诱导剂并解释其中涉及的机制。
