Transcriptional regulation of CYP3A4 and MDR1 mediated via nuclear receptor PXR by helicid

Abstract

Abstract: AIM: To establish nuclear receptor PXR-expressed CYP3A4 and MDR1 dual luciferase reporter gene platform, and analyze the effect of helicid on hPXR-mediated transcriptional regulation of CYP3A4 and MDR1 genes.METHODS: PXR expression plasmid and reporter gene plasmid of either CYP3A4 or MDR1 were constructed based upon the dual luciferase reporter gene platform, and co-transfected to HepG2 cells.The transfected cells were treated with helicid of various concentrations (0.004, 0.04 and 0.4 μmol/L) for 48 hours while rifampin (10 μmol/L) was included as the positive control.Cells were lysized and luciferase activity was determined using a Dual-luciferase reporter assay kit (Promega, Madison, WI).RESULTS: Helicid, at tested concentrations, did not significantly increase promoter activities of CYP3A4 and MDR1 in HepG2 cells transfected with PXR expression plasmid (P >0.05).CONCLUSION: Nuclear receptor PXR-expressed CYP3A4 and MDR1 dual luciferase reporter gene platforms were successfully established, and helicid does not significantly induce the transcription of CYP3A4 and MDR1.

Key words: helicid, human pregnant X receptor, CYP3A4, MDR1

CLC Number:

XIE Hai-tang, WANG Guo, FAN Lan, ZHANG Li-sheng, DAI Xiao-chang, ZHOU Honghao. Transcriptional regulation of CYP3A4 and MDR1 mediated via nuclear receptor PXR by helicid[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2008, 13(12): 1416-1421.

References

[1] Conney AH.Induction of microsomal enzymes by foreign chemicals and carcinogenesis by polycyclic aromatic hydrocarbons:G.H.A.Clowes Memorial Lecture [J]. Cancer Res, 1982, 42(12):4875-4917.
[2] Bertilsson G, Heidrich j, Svensson K, et al.Identification of a human nuclear receptor defines a new signaling pathway for CYP3A induction[J].Proc Natl Acad Sci USA, 1998, 95(21):12208-12213.
[3] Blumberg B, Sabbagh WJ, Juguilon H, et al.SXR, a novel steroid and xenobiotic-sensing nuclear receptor[J]. Genes Dev, 1998, 12(20):3195-3205.
[4] Kliewer SA, Moore JT, Wade L, et al.An orphan nuclear receptor activated by pregnanes defines a novel steroid signaling pathway[J].Cell, 1998, 92(1):73-82.
[5] Honkakoski P, Zelko I, Sueyoshi T, et al.The nuclear orphan receptor CAR-retinoid X receptor heterodimer activates the phenobarbital-responsive enhancer module of the CYP2B gene[J].Mol Cell Biol, 1998, 18(10):5652-5658.
[6] Sueyoshi T, Kawamoto T, Zelko I, et al.The repressed nuclear receptor CAR responds to phenobarbital in activating the human CYP2B6 gene[J].J Biol Chem, 1999, 274(10):6043-6046.
[7] Moore LB, Parks DJ, Jones SA, et al.Orphan nuclear receptors constitutive androstane receptor and pregnane X receptor share xenobiotic and steroid ligands[J].J Biol Chem, 2000, 275(20):15122-15127.
[8] Lehmann JM, Mckee DD, WatsonMA, et al.The human orphan nuclear receptor PXR is activated by compounds that regulate CYP3A4 gene expression and cause drug interactions[J].J Clin Invest, 1998, 102(5):1016-1023.
[9] Goodwin B, Hodgson E, Liddle C.The orphan human pregnane X receptor mediates the transcriptional activation of CYP3A4 by rifampicin through a distal enhancer module [J].Mol Pharmacol, 1999, 56(6):1329-1339.
[10] Greiner B, Eichelbaum M, Fritz P, et al.The role of intestinal P-glycop rotein in the interaction of digoxin and rifampin[J].J Clin Invest, 1999, 104(2):147-153.
[11] Geick A, Eichelbaum M, Burk O.Nuclear recep tor response elements mediate induction of intestinalMDR1 by rifampin[J].J B iol Chem, 2001, 276(18):14581-14587.
[12] Yi W, Cao R, Wen H, et al.Bioorg synthesis and biological evaluation of helicid analogues as mushroom tyrosinase inhibitors[J].Med Chem Lett, 2008, 18(24):6490-6493.
[13] Wen H, Lin C, Que L, et al.Synthesis and biological evaluation of helicid analogues as novel acetylcholinesterase inhibitors [J].Eur J Med Chem, 2008, 43(1):166-173.
[14] 杨蓉, 肖涵, 戴晓畅.豆腐果苷抗抑郁作用研究[J].中药药理与临床, 2007, 23(6):22-32.
[15] 童九翠, 孙瑞元, 江思艳, 等.豆腐果苷衍生物W0620 实验性抗抑郁作用量效关系的研究[J].中国临床药理学与治疗学, 2008;13(11):1277-1281.
[16] 豆腐果苷分散片[J].中国医药技术与市场, 2007, 7(1):48-49.
[17] Hu DL, Wang G, Li Z, et al.The Establishment of in vitro screening system for CYP3A4 and CYP2B6 inducers [J].China Medical Engineering, 2007, 15(8):646-649.
[18] Geick A, Eichelbaum M, Burk O.Nuclear receptor response elements mediate induction of intestinal MDR1 by rifampin[J].J Biol Chem, 2001, 276(18):14581-14587.
[19] Chen H, Lin RJ, Xie W, et al.Regulation of hormone induced histone hyperacetylation and gene activation via acetylation of an acety lase[J].Cell, 1999, 98(5):675-686.
[20] Xie W, Barwick JL, Simon CM, et al.Humanized xenobiotic response in mice expressing nuclear receptor SXR [J].Nature, 2000, 406(6794):435-439.
[21] Staudinger JL, Goodwin B, Jones SA, et al.The nuclear receptor PXR is a lithocholic acid sensor that protects against liver toxicity[J].Proc Natl Acad Sci USA, 2001, 98(6):3369-3374.
[22] Eichelbaum M, Fromm MF, Schwab M.Clinical aspects of the MDR1 (ABCB1) gene polymorphism[J].Ther Drug Monit, 2004, 26(2):180-185.
[23] 刘晓明, 李杰, 闫凤兰, 等.精神分裂症与抑郁症凶杀行为相关因素分析[J].中国民康医学, 2008, 20(4): 882.
[24] 凌丽, 蒋萌.抑郁症的病因探索及治疗概述[J].时珍国医国药, 2007, 18(12):5313-5314.
[25] 顾晓燕.几类常见抗抑郁症药物的特点[J].科技信息, 2008, 21:575.
[26] Hao K, Cao YG, Zhao YN, et al.Bioequivalence of two tablet formulations of helicidum adminstered in single dose to healthy Chinese volunteers[J].Arzneimittelforschung, 2007, 57(8):522-525.