基于孕烷受体-细胞色素P4503A4通路筛选复方丹参中的效应成分实验研究  

Screening Active Components in Compound Danshen Based on PXR-CYP3A4:an Experimental Study

作　　者：肖勇[1,2] 马增春[2] 王宇光[2] 谭洪玲[2] 刘浩生[2] 张娴勰[2] 陆倍倍[2] 梁乾德[2] 汤响林[2] 肖成荣[2] 高月[1,2] 

机构地区：[1]中南大学药学院,长沙410013 [2]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《中国中西医结合杂志》2014年第5期606-610,共5页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家973计划项目(No.2012CB518402;No.2011CB505304);国家自然科学基金资助项目(No.81274127);北京市自然科学基金资助项目(No.7112110)

摘　　要：目的采用孕烷受体-细胞色素P450 3A4(pregnane X receptor-cytochrome P450 3A4,PXRCYP3A4)工程细胞株筛选复方丹参中诱导或抑制PXR-CYP3A4通路的化学成分。方法利用实验室构建的PXR-CYP3A4稳定转染人肝母细胞癌(human hepatoblastoma G2,HepG2)工程细胞株结合报告基因技术,筛选复方丹参中诱导或抑制PXR-CYP3A4通路的化学成分,并从酶活性水平进行确证。实验分为阳性对照组(RIF,10μmol/L)、溶剂对照组(DMSO,0.1%)、药物处理各剂量组(人参皂苷Rc、Rf、Rb2、Rg2、F2、F1、Re,丹参酮Ⅰ,异龙脑5、10、25、50、100、200μmol/L),每种药物设置6个复孔,加药36、48及60h后吸取细胞培养液上清,测定CYP3A4活性,计算诱导倍数。结果与溶剂对照组比较,人参皂苷Rc、Rf、Rb2、Rg2、F2、F1、Re,丹参酮Ⅰ,异龙脑50、100μmol/L处理30、48和60h时,诱导倍数均升高(P<0.05),其中药物浓度200μmol/L组处理48和60h时,人参皂苷Rb2、Rg2、F1组诱导倍数均显著高于RIF阳性对照组(P<0.05)。人参皂苷48h时Rc、Rf、Rb2、F2、F1组CYP3A4酶活性提高(P<0.05)。结论复方丹参中的人参皂苷Rc、Rf、Rb2、F2、F1,丹参酮Ⅰ及异龙脑可以诱导CYP3A4酶。Objective To screen active components in Compound Danshen （CD） based on pregnane X receptor-cytochrome P450 3A4 （PXR-CYP3A4）. Methods By using PXR-CYP3A stable transfection human hepatoblastoma G2 （HepG2） cell lines engineering cell strain combined reporter genes technology, active components that induce or inhibit PXR-CYP3A4 paths in CD were screened, and confirmed at the level of enzymic activities. The experiment was divided into the positive control group （RIF 10μmol/L）, the DMSO group （DMSO 0.1%）, each dose of treatment groups （ginsenoside Rc, Rf, Rb2, Rg2, F2, F1, tanshinone I , isoborneol 5, 10,25, 50, 100, and 200 μmol/L; each with six duplicates）. Cells medium was removed 36, 48, and 60 h after treatment. The activity of CYP3A4 was then determined in the supernant and the fold induction was calculated. Results Compared with the DMSO group, the fold induction increased when ginsenoside Rc, Rf, Rb2, Rg2, F2, F1, tanshinone I , and isoborneol 50 and 100 μmol/L was respectively intervened for 36, 48, and 60 h （P 〈0.05）. When cells were treated with isoborneol 200 μmol/L for 48 and 60 h,the fold induction of ginsenoside Rb2, Rg2, and F1 was significantly higher than that of the RIF group （P 〈0.05）. Enzymic activity results showed that ginsenoside Rc, Rf, Rb2, F2, and F1 could increase the enzyme activity of CYP3A4 at 48 h （P 〈0.05）. Conclusion Ginsenoside Rc, Rf, Rb2, F2, F1, tanshinone I , and isoborneol in DC could induce CYP3A4 enzymes.

关 键 词：细胞色素P450 3A4 孕烷受体 复方丹参 相互作用 

分 类 号：R735.7[医药卫生—肿瘤]