[Establishment of in vitro evaluation model for CYP2B6 induction and its application to screen inducers among TCMs]

Yao Xue Xue Bao. 2013 Jan;48(1):119-24.
[Article in Chinese]

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aryl Hydrocarbon Hydroxylases* / genetics
  • Aryl Hydrocarbon Hydroxylases* / metabolism
  • Constitutive Androstane Receptor
  • Cytochrome P-450 CYP2B6
  • Drugs, Chinese Herbal / isolation & purification
  • Drugs, Chinese Herbal / pharmacology*
  • Genes, Reporter
  • Genetic Vectors
  • Hep G2 Cells
  • High-Throughput Screening Assays
  • Humans
  • Luciferases / genetics
  • Luciferases / metabolism
  • Oximes / pharmacology
  • Plants, Medicinal / chemistry*
  • Plasmids
  • Pregnane X Receptor
  • Receptors, Cytoplasmic and Nuclear* / genetics
  • Receptors, Cytoplasmic and Nuclear* / metabolism
  • Receptors, Steroid* / genetics
  • Receptors, Steroid* / metabolism
  • Rifampin / pharmacology
  • Thiazoles / pharmacology
  • Transfection

Substances

  • Constitutive Androstane Receptor
  • Drugs, Chinese Herbal
  • Oximes
  • Pregnane X Receptor
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Steroid
  • Thiazoles
  • 6-(4-chlorophenyl)imidazo(2,1-b)(1,3)thiazole-5-carbaldehyde O-(3,4-dichlorobenzyl)oxime
  • Luciferases
  • Aryl Hydrocarbon Hydroxylases
  • CYP2B6 protein, human
  • Cytochrome P-450 CYP2B6
  • Rifampin