Abstract
The effects of toluene in dimethylformamide (DMF)-induced hepatotoxicity were investigated with respect to the induction of cytochrome P-450 (CYP) and the activities of related enzymes. The rats were treated intraperitoneally with the organic solvents in olive oil (Single treatment groups: 450 [D1], 900 [D2], 1,800 [D3] mg DMF, and 346 mg toluene [T] per kg of body weight; Combined treatment groups: D1+T, D2+T, and D3+T) once a day for three days, while the control group received just the olive oil. Each group consisted of 4 rats. The activities of the xenobiotic metabolic enzymes and the hepatic morphology were assessed. The immunoblots indicated that the expression of CYP2E1 was considerably enhanced depending on the dosage of DMF and the CYP2E1 blot densities were significantly increased after treatment with both DMF and toluene, compared to treatment with DMF alone. The activities of glutathione- S-transferase and glutathione peroxidase were either decreased or remained unaltered after treatment with DMF and toluene, whereas the lipid peroxide levels were increased with increasing dosage of DMF and toluene. The liver tissue in the D3 group (1,800 mg/kg of DMF) showed signs of microvacuolation in the central vein region and a large necrotic zone around the central vein, in rats treated with both DMF (1,800 mg/kg) and toluene (D3T). These results suggest that the expression of CYP2E1 is induced by DMF and enhanced by toluene. These changes may have facilitated the accelerated formation of N-methylformamide (NMF) from toluene, and the generated NMF may directly induce liver damage.
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References
IPCS. (1991) Dimethylformamide. Environmental Health Criteria 114. WHO, Geneva, pp. 21–23.
Lauwerys, R.R., Kivits, A., Lhoir, M., Rigolet, P., Houbeau, D., Buchet, J.P. and Roels, H.A. (1980) Biological surveillance of workers exposed to dimethylformamide and the influence of skin protection on its percutaneous absorption. Int. Arch. Occup. Environ. Health, 45, 189–203.
Kennedy, G.L. (2012) Toxicology of dimethyl and monomethyl derivatives of acetamide and formamide: a second update. Crit. Rev. Toxicol., 42, 793–826.
Amato, G., Longo, V., Mazzaccaro, A. and Gervasi, P.G. (1996) Microsomal oxidation of N,N-diethylformamide and its effect on P-450-dependent monooxygenase in rat liver. Chem. Res. Toxicol., 9, 882–890.
Kim, K.W., Choi, B.S., Kang, S.K. and Moon, Y.H. (1999) A study on the N-methylformamide excretion rate of workers at synthetic leather factories in Korea. Korean J. Occup. Environ. Med., 11, 106–112.
Koh, S.B., Cha, B.S., Kim, K.W., Kang, S.K. and Jeong, H.S. (1999) Induction of hepatic microsomal cytochrome P-450 by N,N-dimethylformamide in Sprague-Dawley rats. Korean J. Prev. Med., 32, 88–94.
Pearson, P.G., Slatter, J.G., Rashed, M.S., Han, D.H. and Baillie, T.A. (1991) Carbamoylation of peptides and proteins in vitro by S-(N-methylcarbamoyl) glutathione and S-(Nmethylcarbamoyl)- cysteine, two electrophilic S-linked conjugates of methyl isocyanate. Chem. Res. Toxicol., 4, 436–444.
Mráz, J. and Nohová, H. (1992) Absorption, metabolism and elimination of N,N-dimethyl-formamide in humans. Int. Arch. Occup. Environ. Health, 64, 85–92.
Gescher, A. (1993) Metabolism of N,N-dimethylformamide: key to the understanding of its toxicity. Chem. Res. Toxicol., 6, 245–251.
Chang, H.Y., Yun, Y.D., Yu, Y.C., Shih, T.S., Lin, M.S., Kuo, H.W. and Chen, K.M. (2005) The effects of simultaneous exposure to methyl ethyl ketone and toluene on urinary biomarkers of occupational N,N-dimethylformamide exposure. Toxicol. Lett., 155, 385–395.
Wrbizky, R. (1999) Liver function in workers exposed to N,Ndimethylformamide during the production of synthetic textiles. Int. Arch. Occup. Environ. Health, 72, 19–25.
Van den Bulcke, M., Rosseel, M.T., Wijnants, P., Buylaert, W. and Belpaire, F.M. (1994) Metabolism and hepatotoxicity of N,N-dimethylformamide, N-hydroxymethyl-N-methylformamide, and N-methylformamide in the rat. Arch. Toxicol., 68, 291–295.
ACGIH. (2011) Threshold limit values for chemical substances and physical agents and biological exposure indices. ACGIH, Cincinnati, Ohio, pp. 27–57.
Wang, D.H., Horike, T., Mizuuchi, H., Ishii, K., Zhen, L.X. and Taketa, K. (1996) Liver function tests of workers exposed to toluene and toluene/dimethylformamide at low concentrations. J. Occup. Health, 38, 113–117.
Kim, T.H., Kim, Y.W., Shin, S.M., Kim, C.W., Yu, I.J. and Kim, S.G. (2010) Synergistic hepatotoxicity of N,N-dimethylformamide with carbon tetrachloride in association with endoplasmic reticulum stress. Chem. Biol. Interact., 184, 492–501.
Kim, K.W., Kim, H.Y., Park, S.S., Jeong, H.S., Park, S.H., Lee, J.Y., Jeong, J.H. and Moon, Y.H. (1999) Gender differences in activity and induction of hepatic microsomal cytochrome P-450 by 1-bromopropane in Sprague-Dawley rats. J. Biochem. Mol. Biol., 32, 232–238.
Lowry, O.H., Rosebrough, N.J., Farr, A.L. and Randall, R.J. (1951) Protein measurement with Folin phenol reagent. J. Biol. Chem., 193, 265–275.
Omura, T. and Sato, R. (1964) The carbon monooxide-binding pigment of liver microsomes. I. evidence for its hemoprotein nature. J. Biol. Chem., 239, 2370–2378.
Burke, M.D. and Meyer, R.T. (1975) Inherent specificities of purified cytochrome P-450 and P-448 toward biphenyl hydroxylation and ethoxyresorufin deethylation. Drug Metab. Dispos., 3, 245–253.
Lubet, R.A., Meyer, R.T., Cameron, J.W., Nims, R.W., Burke, M.D., Wolff, T. and Guengerich, F.P. (1985) Dealkylation of Hepatotoxicity of Dimethylformamide and Toluene in Rats 193 pentoxyresorufin: A rapid and sensitive assay for measuring induction of cytochrome (s) P-450 by phenobarbital and other xenobiotics in the rat. Arch. Biochem. Biophys., 238, 43–48.
Koop, D.R. (1986) Hydroxylation of ρ-nitrophenol by rabbit ethanol-inducible cytochrome P-450 isozymes 3a. Mol. Pharmacol., 29, 399–404.
Yagi, K. (1987) Lipid peroxide and human diseases. Chem. Phys. Lipids, 45, 337–351.
Habig, W.H., Pabest, M.J. and Jakoby, W.B. (1974) Glutathione S-transferase. The first enzymatic step in mercapturic acid formation. J. Biol. Chem., 249, 7130–7139.
Tappel, A.L. (1978) Glutathione peroxidase andhydroperoxidase; in Methods in Enzymology. Academic Press, New York, pp. 506–513.
Laemmli, U.K. (1970) Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.
Kim, K.W., Chang, S.K., Jeong, H.Y., Lee, J.Y., Moon, Y.H. and Park, S.S. (1997) Induction of hepatic microsomal cytochrome P-450 by styrene in rat. Korean J. Occup. Med., 9, 604–613.
Klotz, A.V., Stegeman, J.J. and Walsh, C. (1984) An alternative 7-ethoxyresorufin O-deethylase activity assay: A continuos visible spectrophotometric methods for measurement of cytochrome P-450 monooxygenase activity. Anal. Biochem., 140, 138–145.
Spatzenegger, M. and Jaeger, W. (1995) Clinical importance of hepatic cytochrome P450 in drug metabolism. Drug Metab. Rev., 27, 397–417.
Kim, K.W., Park, S.S., Kim, T.K., Moon, Y.H. and Chang, S.K. (1996) Combinatory effects of benzene, toluene and xylene on the induction of rat liver microsomal enzymes and their metabolites. Toxicol. Res., 12, 9–15.
Takahashi, J., Furuhata, Y., Ikeda, A., Takahashi, M., Iwata, H., Kazusaka, A. and Fujita, S. (1999) Characterization of hepatic cytochrome P450 isozyme composition in the transgenic rat expressing low level human growth hormone. Xenobiotica, 29, 1203–1212.
Petrulis, J.R. and Bunce, N.J. (2000) Competitive behavior in the interactive toxicology of halogenated aromatic compounds. J. Biochem. Mol. Toxicol., 14, 73–81.
Porter, T.D. and Coon, M.J. (1991) Cytochrome P-450: multiplicity of isoforms, substrates and catalytic and regulatory mechanisms. J. Biol. Chem., 266, 13469–13472.
Guengerich, F.P., Kim, D.H. and Iwasaki, M. (1991) Role of cytochrome P-4502E1 in the oxidation of many low molecular weight cancer suspects. Chem. Res. Toxicol., 4, 168–179.
Nakajima, T. (1997) Cytochrome P450 isoforms and the metabolism of volatile hydrocarbons of low relative molecular mass. J. Occup. Health, 39, 83–91.
Hyland, R., Gescher, A., Thummel, K., Schiller, C., Jheeta, P., Mynett, K., Smith, A.W. and Mráz, J. (1992) Metabolic oxidation and toxification of N-methylformamide catalyzed by the cytochrome P450 isozyme CYP2E1. Mol. Pharmacol., 41, 259–266.
Chieli, E., Saviozzi, M., Menicagli, S., Branca, T. and Gervasi, P.G. (1995) Hepatotoxicity and P-4502E1-dependent metabolic oxidation of N,N-dimethylformamide in rats and mice. Arch. Toxicol., 69, 165–170.
Kestell, P., Threadgill, M.D., Gescher, A., Gledhill, A.P., Shaw, A.J. and Farmer, P.B. (1987) An investigation of the relationship between hepatotoxicity and the metabolism of Nalkyl-formamide. J. Pharmacol. Exp. Ther., 240, 265–270.
Imazu, K., Fujishiro, K. and Inove, N. (1992) Effects of dimethylformamide on hepatic microsomal monooxygenase system and glutathione metabolism in rats. Toxicology, 72, 41–50.
Kim, T.H. and Kim, S.G. (2011) Clinical outcomes of occupational exposure to n,n-dimethylformamide: perspectives from experimental toxicology. Saf. Health Work, 2, 97–104.
Kennedy, G.L. Jr. (1986) Biological effects of acetamide, formamide, and their monomethyl and dimethyl derivatives. Crit. Rev. Toxicol., 17, 129–182.
Lora, E.F., Stuart, L.S. and Carrie, A.R. (1990) Liver injury in workers exposed to dimethylform-amide. Scand. J. Work Environ. Health, 16, 289–92.
Wang, J.D., Lai, M.Y., Chen, J.S., Lin, J.M., Chiang, J.R., Shiau, S.J. and Chan, W.S. (1991) Dimethylformamideinduced liver damage among synthetic leather workers. Arch. Environ. Health, 46, 161–166.
Church, D.F. and Pryor, W.A. (1985) Free-radical chemistry of cigarette smoke and its toxicological implications. Environ. Health Perspect., 64, 111–126.
Wurzel, H., Yeh, C.C., Gairola, C. and Chow, C.K. (1995) Oxidative damage and antioxidant status in the lungs and bronchoalveolar lavage fluid of rats exposed chronically to cigarette smoke. J. Biochem. Toxicol., 10, 11–17.
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Kim, KW., Chung, Y.H. Hepatotoxicity in Rats Treated with Dimethylformamide or Toluene or Both. Toxicol Res. 29, 187–193 (2013). https://doi.org/10.5487/TR.2013.29.3.187
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DOI: https://doi.org/10.5487/TR.2013.29.3.187