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Archives of Toxicology

, Volume 77, Issue 10, pp 555–560 | Cite as

Inter-species comparisons of hepatic cytochrome P450 enzyme levels in male ruminants

  • Miroslav MachalaEmail author
  • Pavel Souček
  • Jiří Neča
  • Robert Ulrich
  • Jiří Lamka
  • Barbora Szotáková
  • Lenka Skálová
Toxicokinetics and Metabolism

Abstract

Our current knowledge about the biotransformation enzymes in wild ruminants is limited. The present study aimed to compare basic levels and specific activities of cytochrome P450 isoforms (CYP1A, 2A, 2B, 2C, 2D, 2E, 3A, 4A) in males of red deer (Cervus elaphus), fallow deer (Dama dama), roe deer (Capreolus capreolus) and mouflon (Ovis musimon). The proteins from the major cytochrome P450 (CYP) subfamilies were detected in all ruminant species by Western blotting, using polyclonal antibodies raised against rat or human CYP enzymes. The immunochemical data seem to suggest that humans and wild ruminants share some similar hepatic CYP enzymes corresponding to members of subfamilies 2 and 3; ruminant liver samples also contained two proteins cross-reacting with anti-rat CYP1A antibodies. High activities of CYP1A enzymes found in liver microsomes of male fallow deer and roe deer are indicative of increased susceptibility of these species towards promutagens that are metabolically activated by these CYPs. On the other hand, low activities of CYP1A-dependent alkoxyresorufin O-dealkylase activities were detected in male mouflons. Oxidative metabolism of testosterone was significantly higher in wild ruminants than the values previously reported from bulls. Androstene-3,17-dione and 6β-hydroxytestosterone were the most important products of testosterone oxidation in liver microsomes of all the ruminant species under study. The highest CYP3A-dependent testosterone 6β-hydroxylase activity was found in mouflons and fallow deer. A different pattern of CYP activities towards testosterone was found in roe deer, which showed high activities of testosterone 2β-hydroxylase and lower production of androstene-3,17-dione. An increased activity of CYP4A-dependent laurate 12-hydroxylase found in roe deer and mouflons might indicate a higher metabolic turnover of fatty acids. The data on CYP activities indicated that high metabolic rates of steroids, fatty acids, and xenobiotics may occur in male wild ruminants. The highest hepatic activities specific for CYP3A, CYP2C, CYP2D, and CYP2E enzymes were found in mouflon, suggesting that this species has the highest biotransformation capacity.

Keywords

CYPs Ruminants Immunoblotting Biotransformation 

Notes

Acknowledgements

The authors wish to express their sincere thanks to Prof. F.P. Guengerich for his generous gifts (antisera against human CYPs, purified human CYP enzymes and expressing clones) and to Mrs. Eva Grobnerová and Marie Gájová (VRI, Brno) for their excellent technical assistance. This work was supported by the Grant Agency of the Czech Republic (grant no. 524/00/0514), Czech Ministry of Agriculture (grant no. QC0194) and the Grant Agency of Charles University (grant no. 96/2002/C/FaF). The authors hereby declare that the experiments comply with the current laws of the Czech Republic.

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Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Miroslav Machala
    • 1
    Email author
  • Pavel Souček
    • 2
  • Jiří Neča
    • 1
  • Robert Ulrich
    • 1
  • Jiří Lamka
    • 3
  • Barbora Szotáková
    • 3
  • Lenka Skálová
    • 3
  1. 1.Veterinary Research InstituteBrnoCzech Republic
  2. 2. National Institute of Public HealthPragueCzech Republic
  3. 3.Faculty of PharmacyCharles UniversityHradec KrálovéCzech Republic

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