硝苯地平对人孕烷X受体介导的CYP2B6和CYP2C9的转录调节作用  

Transcriptional regulation effects of Nifedipine for CYP2B6 and CYP2C9 mediated by hPXR

作　　者：胡东莉[1] 王果[1] 李智[1] 刘昭前[1] 周宏灏[1] 

机构地区：[1]中南大学临床药理研究所,湖南长沙410078

出　　处：《中国现代医学杂志》2008年第16期2307-2311,共5页China Journal of Modern Medicine

基　　金：中南大学博士后基金资助

摘　　要：目的建立hPXR介导的CYP2B6、CYP2C9药物诱导剂的体外筛选体系,分析硝苯地平诱导CYP2B6、CYP2C9的分子机制。方法利用双荧光素酶报告基因系统,将包含hPXR蛋白识别和结合调控元件的CYP2B6和2C9启动子序列插入报告基因上游,将表达载体和报告载体共转染HepG2细胞,以10μM利福平为阳性对照,用1、5或10μM硝苯地平处理48h后裂解细胞进行双荧光素酶活性检测;或用10μM硝苯地平分别处理细胞12、24和48h后进行双荧光素酶活性检测。结果硝苯地平激活hPXR的起始浓度为1～5μm,在5μm浓度下,硝苯地平诱导CYP2B6和2C9表达增加3.93和3.59倍,与DMSO溶媒组差异存在显著性(P<0.001),在10μm硝苯地平作用下,CYP2B6和2C9表达分别增加6.23和5.24倍,与溶媒对照组和5μm浓度组差异都存在显著性(P<0.001)。10μm硝苯地平处理12h后即能显著地诱导CYP2B6和CYP2C9表达增强,但其诱导倍数与24和48h组差异存在显著性(P<0.001)。结论该研究成功构建了hPXR介导的CYP2B6和CYP2C9药物诱导剂的体外筛选体系;硝苯地平通过激活hPXR介导了CYP2B6和2C9的表达上调,并表现出明确的剂量-效应关系。[Objective] To establish an in vitro system to screen drug inducers of CYP2B6 and CYP2C9 enzymes, and to analyze the molecular mechanism of induce effect of Nifedipine for CYP2B6 and CYP2C9 enzymes. [Method] Clonging the promoters of CYP2B6 and CYP2C9 which contain the elements that hPXR, a kind of nuclear receptor, can recognize and bind to. The promoter elements were inserted to the upstream of firefly luciferase reporter gene in pGL4.17 vector. The reporter vectors and hPXR expression vector were co-transfected to HepG2 cell line, and dual-luciferase activity was analyzed after the cell treated with different concentration nifedipine and different treatment times. [Result]The fold activation value of Nifedipine treatment group had statistical significant difference from DMSO group at the concentration of 5 μm and 10 μm（P 〈0.001）. 10 μm Nifedipine can significantly induce CYP2B6 and CYP2C9 after 12 h, 24 h and 48 h treatment, but there was no statistical significant difference between 24 h and 48 h treatment group. [Conclusion] We successfully established an in vitro system which can be used for screen the drug agonist for hPXR and increase the expression of CYP2B6 and CYP2C9 enzymes. The inductive effects of Nifedipine for CYP2B6 and CYP2C9 can be mediated by hPXR.

关 键 词：硝苯地平 hPXR CYP286 CYP2C9 

分 类 号：R972[医药卫生—药品]