Coordinate regulation of UDP-glucuronosyltransferase UGT1A6 induction by 3-methylcholanthrene and multidrug resistance protein MRP2 expression by dexamethasone in primary rat hepatocytes

Biochem Pharmacol. 2002 Jun 15;63(12):2137-44. doi: 10.1016/s0006-2952(02)01022-5.

Abstract

Concentration-dependent regulation of 3-methylcholanthrene (MC) inducibility of UDP-glucuronosyltransferase UGT1A6 by the synthetic glucocorticoid, dexamethasone (DEX) was studied. Treatment of cultured rat hepatocytes with MC, 0.1, 1, and 10 microM DEX, and MC combined with DEX, resulted in different induction patterns measured in the intact cells compared to that observed in the microsomes prepared from the same cells. DEX treatment in various concentrations caused a concentration-dependent increase in p-nitrophenol (p-NP) conjugation in intact cells (3-, 4-, and 5-fold over control, respectively), and it positively regulated MC induction (4-, 5-, and 6-fold over control, respectively). In contrast, DEX had smaller effect on microsomal p-NP conjugation (115, 200, 220% of control, respectively) and although MC induction was increased significantly by 0.1 microM DEX (520% of control), but higher concentrations of DEX (10 microM) decreased the degree of induction to 410%. Similar results obtained from in vivo experiments showed that at high DEX concentration (100mg/kg), the rate of MC induction (540%) decreased (420%). Permeabilization of the plasma membrane resulted in a 15-fold increase of p-NP conjugation indicating the importance of transport in the rate of overall p-NP elimination, and the induction pattern was similar to that observed in microsomes isolated from cells. Hyper-osmolarity (405 mOsmol/L) led to a 3-fold decrease of p-NP conjugation, the loss of DEX inducibility and reduction of the MRP2 protein level. Our results suggest coordinated regulation of UGT1A6 inducibility and substrate or product transport by DEX.

MeSH terms

  • Animals
  • Anti-Inflammatory Agents / pharmacology
  • Blotting, Western
  • Carcinogens / pharmacology
  • Cells, Cultured
  • Dexamethasone / pharmacology*
  • Enzyme Induction / drug effects
  • Glucuronosyltransferase / biosynthesis*
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism
  • Male
  • Methylcholanthrene / pharmacology*
  • Mitochondrial Proteins*
  • Rats
  • Rats, Wistar
  • Ribosomal Proteins / biosynthesis*
  • Saccharomyces cerevisiae Proteins*

Substances

  • Anti-Inflammatory Agents
  • Carcinogens
  • MRP2 protein, S cerevisiae
  • Mitochondrial Proteins
  • Ribosomal Proteins
  • Saccharomyces cerevisiae Proteins
  • Methylcholanthrene
  • Dexamethasone
  • UDP-glucuronosyltransferase, UGT1A6
  • Glucuronosyltransferase