Induction of Microsomal Epoxide Hydrolase, rGSTA2, rGSTA3/5, and rGSTM1 by Disulfiram, but not by Diethyldithiocarbamate, a Reduced Form of Disulfiram

  • Kim, Sang-Geon (College of Pharmacy, Duksung Women's University, 419 Ssangmoon-dong, Dobong-gu Seoul 132-714) ;
  • Kim, Hye-Jung (College of Pharmacy, Duksung Women's University, 419 Ssangmoon-dong, Dobong-gu Seoul 132-714)
  • Published : 1997.12.01


Disulfiram (DSF) and diethyldithiocarbamate (DDC), a reduced form of DSF, protect the liver against toxicant-induced injury through inhibition of cytochrome P450 2E1. The effect of DSF and DDC on the levels of major hepatic microsomal epoxide hydrolase (mEH) and glutathione S-transferase (GST) expression was comparatively studied, given the view that these enzymes are involved in terminal detoxification events for high energy intermediates of xenobiotics. Treatment of rats with a single dose of DSF (20-200 mg/kg, po) resulted in 2- to 15-fold increases in the mEH mRNA level at 24 hr with the ED$_{50}$ value being noted as 60 mg/kg. The mEH mRNA level was elevated ~15-fold at 24 hr after treatment at the dose of 100 mg/kg, whereas the hepatic mRNA level was rather decreased from the maximum at the dose of 200 mg/kg, indicating that DSF might cause cytotoxicity at the dose. In contrast to the effect of DSF, DDC only minimally elevated the mEH mRNA level at the doses employed. DSF moderately increased the major GST mRNA levels in the liver as a function of dose, resulting in rGSTA2, rGSTA3/5 or rGSTM1 mRNA levels being elevated 3- to 4-fold at 24 hr post-treatment, whereas the rGSTM2 mRNA level was not altered. DDC, however, failed to stimulate the mRNA levels for major GST subunits, indicating that the reduced form of DSF was ineffective in stimulating the GST the expression. The effect of other organosulfides including aldrithiol, 2, 2'-dithiobis(benzothiazole) (DTB), tetramethylthiouram disulfide (TMTD) and allyl disulfide (ADS) on the hepatic mEH and GST mRNA expression was assessed in rats in order to further confirm the increase in the gene expression by other disulfides. Treatment of rats with aldrithiol (100 mg/kg, po) resulted in a 16-fold increase in the mEH mRNA level at 24 hr post-treatment. DTB, TMTD and ADS also caused 5-, 9- and 12-fold increases in the rnRNA level, respectively, as compared to control. Thus, all of the disulfides examined were active in stimulating the mEH gene in the liver. The organosulfides significantly increased the rGSTA2, rGSTA3, rGSTA5 and rGSTM1 mRNA levels at 24 hr after administration. In particular, aldrithiol was very efficient in stimulating the rGSTA and rGSTM genes among the disulfides examined. These results provide evidence that DSF and other sulfides effectively stimulate the mEH and major GST gene expression at early times in the liver and that DDC, a reduced form of DSF, was ineffective in stimulating the expression of the genes, supporting the conclusion that reduced form(s) of organosulfur compound(s) might be less effective in inducing the mEH and GST genes through the antioxidant responsive element(s).


  1. Hepatology v.3 Chemoprotective effects of two dithiolthiones and butylhydroxyanisole against carbon tetrachloride and acetaminophen toxicity Ansher, S.S.;Dolan, P.;Bueding, E.
  2. Toxicol. and Appl. Pharmacol. v.108 Effect of disulfiram on hepatic P4502E1, other microsomal enzymes and hepatotoxicity in rats Brady, J.F.;Xiao, F.;Wang, M.H.;Li, Y.;Ning, S.M.;Gapac, J.M.;Yang, C.S.
  3. Toxicol. Appl. Pharmacol. v.135 Induction of phase I and Phase II drug-metabolizing enzyme mRNA, protein and activity by BHA, ethoxyquin and oltipraz Buetler, T.M.;Gallagher, E.P.;Wang, C.;Stahl, D.L.;Hayes, J.D.;Eaton, D.L.
  4. Crit. Rev. Biochem. Mol. Biol. v.28 Glutathione S-transferases:gene structure and regulation of expression Daniel, V.
  5. The Phamacological Basis of Therapeutics Gilman, A.G.;Rall, T.W.;Nies, A.S.;Taylor, P.
  6. Crit. Rev. Biochem. Mol. Biol. v.30 The glutathione S-transferase supergene family : regulation of GST and the contribution of the isoenzymes to chemoprotection and drug resistance Hayes, J.D.;Pulford, D.J.
  7. Biochem. Mol. Biol. v.52 Expression of glutathione S-transfease Ya, Yb1, Yb2, Yc1 and Yc2 and microsomal epoxide hydrolase gene by thiazole, benzothiazole and benzothiadiazole Kim, S.G.;Cho, M.K.
  8. Carcinogenesis v.14 Thiazole and pyrazine induction of microsomal epoxide hydrolase in rats: 2-cyanonethylene oxide hydrosis Kim, S.G.;Kedderis, G.L.;Batra, R.;Novak, R.F.
  9. Mol. Pharmacol. v.42 Transcriptional regulation of rat microsomal epoxide hydrolase gene by imidazole antimycotic agents Kim, S. G.
  10. J. Pharmacol. Exp. Ther. v.277 Molecular mechanism for alkyl sulfide-modulated carbon tetrachloride-induced hepatoxicity : the role of P450 2E1, P450 2B and glutathione S-transferase expression Kim, S.G.;Chung, H.C.;Cho, J.Y.
  11. Biochem. Pharmacol. v.53 Inhibition of cytochrome P450 2E1 expression by 2-(allylthio)pyrazine, a potential chemoprotective agent : hepatoprotective effects Kim, N.D.;Kwak, M.K.;Kim, S.G.
  12. Mol. Pharmacol. v.51 Enhancement of radiation-inducible hepatic glutathione S-transferase Ya, Yb1, Yb2, Yc1 and Yc2 expression by oltipraz : possible role in radioprotection Kim, S.G.;Nam, S.Y.;Kim, J.H.;Cho, C.K.;Yoo, S.Y.
  13. Drug Metab. Dispos. v.25 Gadolinium chloride inhibition of rat hepatic microsomal epoxide hydrolase and glutathione S-transferase gene expression Kim, S.G.;Choi, S.H.
  14. Biochem. Pharmacol. v.45 Regluation of cytochrome P450 2B1/2 genes by diallyl sulfide, disulfiram and other organosulfur compounds in primary cultures of rat hepatocytes Pan, J.;Hong, J.Y.;Li, D.;Schuetz, E.G.;Guzelian, P. S.;Huang, W.;Yang, C.S.
  15. J. Biol. Chem. v.263 Tissue-specific inductin of murine glutathione transferase mRNAs by butylated hydroxyanisole Pearson, W.R.;Reinhart, J.;Sisk, S.C.;Anderson, K.S.;Adler, P.N.
  16. Biotechniques v.8 An improvement of the single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction Rushmore, T.H.;Morton, M.R.;Pickett, C.B.
  17. J. Biol. Chem. v.266 The antioxidant responsive element Rushmore, T.H.;Morton, M.R.;Pickett, C.B.
  18. Fund. Appl. Tox. v.31 Chelation in metal intoxication XXXVIII : effect of structurally different chelating agents in treatment of nickel intoxication in rat Tandon, S.K.;Singh, S.;Jaln, V. K.;Presad, S.
  19. Tox. Appl. Pharamacol. v.139 Disulfiram and diethyldithiocarbamate intoxication affects the storage and release of striatal dopanmine Vaccarl, A.;Saba, P. L.;Ruiu, S.;Collu, M.;Devoto, P.