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The Effect of Copper on the mRNA Expression Profile of Xenobiotic-Metabolizing Enzymes in Cultured Rat H4-II-E Cells

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Abstract

Copper (Cu2+) is an essential element that plays important roles in physiological functions of the body. However, high Cu2+ levels can have toxic implications. This study aims to investigate the constitutive response to Cu2+ exposure of xenobiotic-metabolizing enzymes in cultured rat liver (H4-II-E) cell lines. Rat cells were exposed to copper sulfate (0–500 μM) for 24 h. The effects of Cu2+ on the messenger RNA (mRNA) expressions of phase I and II enzymes and regulatory elements were examined using real-time PCR. Metallothionein mRNA expression was induced in a dose-dependent manner after treatment with Cu2+. mRNA expressions of phase I enzymes such as cytochrome P450 1A1 and 1A2 (CYP1A1 and CYP1A2) were slightly induced after exposure to low concentrations of Cu2+; however, CYP1A1 and CYP1A2 mRNA expressions were significantly downregulated at higher Cu2+ concentrations. These effects corresponded with expression of aryl hydrocarbon receptor mRNA. The mRNA expressions of phase II enzymes were reduced upon exposure to Cu2+. In conclusion, phase I and II enzyme expressions were significantly modulated upon Cu2+ exposure. These results indicated that Cu2+ exposure had toxicological implications for cultured H4-II-E cells.

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References

  1. Linder MC, Hazegh-Azam M (1996) Copper biochemistry and molecular biology. Am J Clin Nutr 63(5):797S–811S

    CAS  PubMed  Google Scholar 

  2. Letelier ME, Faúndez M, Jara-Sandoval J, Molina-Berríos A, Cortés-Troncoso J, Aracena-Parks P, Marín-Catalán R (2009) Mechanisms underlying the inhibition of the cytochrome P450 system by copper ions. J Appl Toxicol 8:695–702

    Article  Google Scholar 

  3. Goldstein S, Czapski G (1986) Mechanisms of the reactions of some copper complexes in the presence of DNA with superoxide, hydrogen peroxide, and molecular oxygen. J Am Chem Soc 108(9):2244–2250

    Article  CAS  PubMed  Google Scholar 

  4. Nakayama SM, Ikenaka Y, Hamada K, Muzandu K, Choongo K, Teraoka H, Mizuno N, Ishizuka M (2011) Metal and metalloid contamination in roadside soil and wild rats around a Pb-Zn mine in Kabwe, Zambia. Environ Pollut 159(1):175–181

    Article  CAS  PubMed  Google Scholar 

  5. Sato M, Kondoh M (2002) Recent studies on metallothionein: protection against toxicity of heavy metals and oxygen free radicals. Tohoku J Exp Med 196:9–22

    Article  CAS  PubMed  Google Scholar 

  6. Darwish WS, Ikenaka Y, Eldaly E, Ishizuka M (2010) Mutagenic activation and detoxification of benzo[a]pyrene in vitro by hepatic cytochrome P450 1A1 and phase II enzymes in three meat-producing animals. Food Chem Toxicol 48:2526–2531

    Article  CAS  PubMed  Google Scholar 

  7. Darwish WS, Ikenaka Y, El-Ghareeb W, Ishizuka M (2010) High expression of the mRNA of cytochrome P450 and phase II enzymes in the lung and kidney tissues of cattle. Animal 4:2023–2029

    Article  CAS  PubMed  Google Scholar 

  8. Braeuning A, Köhle C, Buchmann A, Schwarz M (2011) Coordinate regulation of cytochrome P450 1a1 expression in mouse liver by the aryl hydrocarbon receptor and the beta-catenin pathway. Toxicol Sci 122(1):16–25

    Article  CAS  PubMed  Google Scholar 

  9. Darwish WS, Ikenaka Y, Ohno M, Eldaly E, Ishizuka M (2010) Carotenoids as regulators for inter-species difference in cytochrome P450 1A expression and activity in food producing animals and rats. Food Chem Toxicol 48:3201–3208

    Article  CAS  PubMed  Google Scholar 

  10. Tchounwou PB, Newsome C, Williams J, Glass K (2008) Copper-induced cytotoxicity and transcriptional activation of stress genes in human liver carcinoma (HepG(2)) cells. Met Ions Biol Med 10:285–290

    PubMed Central  PubMed  Google Scholar 

  11. Jönsson ME, Carlsson C, Smith RW, Pärt P (2006) Effects of copper on CYP1A activity and epithelial barrier properties in the rainbow trout gill. Aquat Toxicol 79(1):78–86

    Article  PubMed  Google Scholar 

  12. Campana O, Sarasquete C, Blasco J (2003) Effect of lead on ALA-D activity, metallothionein levels, and lipid peroxidation in blood, kidney, and liver of the toadfish Halobatrachus didactylus. Ecotoxicol Environ Saf 55:116–125

    Article  CAS  PubMed  Google Scholar 

  13. Ikenaka Y, Nakayama SM, Muroya T, Yabe J, Konnai S, Darwish WS, Muzandu K, Choongo K, Mainda G, Teraoka H, Umemura T, Ishizuka M (2012) Effects of environmental lead contamination on cattle in a lead/zinc mining area: changes in cattle immune systems on exposure to lead in vivo and in vitro. Environ Toxicol Chem 31(10):2300–2305

    Article  CAS  PubMed  Google Scholar 

  14. Darwish WS, Ikenaka Y, Ishizuka M (2013) Biological responses of xenobiotic metabolizing enzymes to lead exposure in cultured H4IIE rat cells. Jpn J Vet Res 61:S13–S22

    PubMed  Google Scholar 

  15. Whitlock JP (1999) Induction of cytochrome P4501A1. Annu Rev Pharmacol Toxicol 39:103–125

    Article  CAS  PubMed  Google Scholar 

  16. Korashy H, El-Kadi A (2004) Differential effects of mercury, lead, and copper on the constitutive and inducible expression of aryl hydrocarbon receptor (AHR)-regulated genes in cultured hepatoma Hepa 1c1c7 cells. Toxicology 201:153–172

    Article  CAS  PubMed  Google Scholar 

  17. Kaliman PA, Nikitchenko IV, Sokol OA, Strel’chenko EV (2001) Regulation of heme oxygenase activity in rat liver during oxidative stress induced by cobalt chloride and Hg+2 chloride. Biochemistry 66:77–82

    CAS  PubMed  Google Scholar 

  18. Vakharia DD, Liu N, Pause R, Fasco M, Bessette E, Zhang QY, Kaminsky LS (2001) Effect of metals on polycyclic aromatic hydrocarbon induction of CYP1A1 and CYP1A2 in human hepatocyte cultures. Toxicol Appl Pharmacol 170(2):93–103

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported in part by the Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, which was awarded to M. Ishizuka (No. 24248056 and 24405004); and from the Japanese Society for Promotion of Science (JSPS) to M. Ishizuka (Core to Core program) and W. S. Darwish (No. 23001097). W. S. Darwish is a recipient of a postdoctoral fellowship from JSPS.

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Correspondence to Mayumi Ishizuka.

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Darwish, W.S., Ikenaka, Y., Nakayama, S. et al. The Effect of Copper on the mRNA Expression Profile of Xenobiotic-Metabolizing Enzymes in Cultured Rat H4-II-E Cells. Biol Trace Elem Res 158, 243–248 (2014). https://doi.org/10.1007/s12011-014-9915-9

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  • DOI: https://doi.org/10.1007/s12011-014-9915-9

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