补骨脂素、异补骨脂素、补骨脂二氢黄酮与异补骨脂查尔酮对CYP2B6诱导作用研究  

Inductive effects of psoralen,isopsoralen,bavachin and isobavachalcone cytochrome on P450 2B6

作　　者：李晗[1,2] 王宇光[2] 马增春[2] 谭洪玲[2] 肖成荣[2] 汤响林[2] 张伯礼[1] 高月[2] 

机构地区：[1]天津中医药大学,天津330007 [2]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《中药药理与临床》2017年第1期15-19,共5页Pharmacology and Clinics of Chinese Materia Medica

基　　金："十二五"国家科技重大专项资助项目(2014ZX09304307001-003);"十二五"国家科技重大专项资助项目(2015ZX09501004003-003)

摘　　要：目的:研究补骨脂中4种单体成分补骨脂素、异补骨脂素、补骨脂二氢黄酮与异补骨脂查尔酮对细胞色素P450 2B6在mRNA和蛋白的影响,并初步探讨该影响的作用机制。方法:利用前期实验室构建的表达质粒pc DNA3.1-h CAR、报告基因质粒p GL4.17-CYP2B6与内参质粒pRL-TK共同转染Hep G2细胞,检测补骨脂素、异补骨脂素、异补骨脂查尔酮和补骨脂二氢黄酮对组成型雄甾烷受体(constitutive androstane receptor,CAR)的转录激活效应;并利用Real time PCR及Western Blot技术对补骨脂素、异补骨脂素、补骨脂二氢黄酮和异补骨脂查尔酮的不同浓度处理组进行mRNA水平表达及蛋白水平表达的检测。结果:双荧光素酶报告基因检测结果显示,补骨脂素、异补骨脂素、补骨脂二氢黄酮和异补骨脂查尔酮(5、10、20、50μmol/L)对CAR具有转录活化作用,其中补骨脂素与异补骨脂素对CAR转录活化效应明显;同时补骨脂素、异补骨脂素、补骨脂二氢黄酮和异补骨脂查尔酮(5、10、20、50μmol/L)均能上调CYP2B6 mRNA水平与蛋白表达水平。结论:补骨脂素、异补骨脂素、补骨脂二氢黄酮和异补骨脂查尔酮可以诱导CYP2B6 mRNA与蛋白质水平的表达,这种诱导作用可能是单体成分通过激活CAR信号通路实现。Objective： To evaluate the inductive effect of psoralen,isopsoralen,bavachin and isobavachalcone on CYP2B6,and further validate the role of constitutive androstane receptor（ CAR） in CYP2B6 expression.Methods： Dual luciferase reporter gene system was performed.psoralen,isopsoralen,bavachin and isobavachalcone（ 5,10,20,50 μmol/L） were screened for constitutive androstane receptor activation by reporter gene assays,and CITCO as the positive control.Further with different concentrations of psoralen,isopsoralen,bavachin and isobavachalcone（ 5,10,20,50 μmol/L） treated on LS174 T cells,RNA and total protein were extracted to detect the regulating effect on CYP2B6 mRNA and protein expression with Real time PCR and Western blot technology respectively.Results： Reporter gene screening showed that the psoralen,isopsoralen,isobavachalcone and bavachin can activate CAR and have potential effects on the induction of CYP2B6 enzyme.Meanwhile,the levels of CYP2B6 mRNA and protein expression were increased by psoralen,isopsoralen,bavachin and isobavachalcone in varying degrees.Conclusion： psoralen,isopsoralen,isobavachalcone and bavachin can induct the gene and protein expression through the CAR-mediated CYP2B6 pathway.

关 键 词：补骨脂 组成型雄甾烷受体 CYP2B6 PCR 蛋白质印迹法 药物相互作用 

分 类 号：R285[医药卫生—中药学]