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Abstract

In the present study, we characterized the kinetic parameters of 7-ethoxyresorufin O-deethylation (EROD) and 7-methoxyresorufin O-demethylation (MROD) in hepatic microsomes from entire and castrated male pigs. Validation parameters of an HPLC-based method to analyse EROD and MROD activities are also described. Eadie-Hofstee plot analysis demonstrated a biphasic kinetic of EROD, indicating that at least two forms of cytochrome P450 are involved in this reaction. MROD followed monophasic kinetic, suggesting that a single enzyme, or enzymes with similar affinities, is responsible for the reaction. Inhibitory effects of alpha-naphthoflavone (ANF), ellipticine and furafylline were studied using microsomes from entire and castrated male pigs. ANF is a known inhibitor of both cytochrome P(450) 1A1 and 1A2 (CYP1A1 and CYP1A2); the presence of ANF in the incubations resulted in the inhibition of both EROD and MROD activities in porcine liver microsomes. EROD activities in porcine liver microsomes were also inhibited by selective CYP1A1 inhibitor ellipticine, but not by CYP1A2 inhibitor furafylline. MROD activities were strongly inhibited by ellipticine and to a much lesser extent by furafylline. Further studies are needed to evaluate substrate specificities of porcine CYP1A1 and CYP1A2.

Keywords

Entire male pigs; castrated male pigs; porcine hepatic microsomes; EROD; MROD; HPLC; alpha-naphthoflavone; ellipticine; furafylline

Published in

Sensors
2009, volume: 9, number: 3, pages: 2134-2147
Publisher: MOLECULAR DIVERSITY PRESERVATION INTERNATIONAL-MDPI

SLU Authors

UKÄ Subject classification

Food Science
Animal and Dairy Science
Veterinary Science

Publication identifier

  • DOI: https://doi.org/10.3390/s90302134

Permanent link to this page (URI)

https://res.slu.se/id/publ/61646