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EROD and MROD as Markers of Cytochrome P450 1A Activities in Hepatic Microsomes from Entire and Castrated Male Pigs
Department of Food Science, Swedish University of Agricultural Sciences, SE-750 07, Uppsala, Sweden
University of South Bohemia, Ceske Budejovice, Research Institute of Fish Culture and Hydrobiology, Zatisi 728, 389 25 Vodňany, Czech Republic
* Author to whom correspondence should be addressed.
Received: 21 February 2009; in revised form: 18 March 2009 / Accepted: 19 March 2009 / Published: 23 March 2009
Abstract: In the present study, we characterized the kinetic parameters of 7-ethoxy-resorufin 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 α-naphthoflavone (ANF), ellipticine and furafylline were studied using microsomes from entire and castrated male pigs. ANF is a known inhibitor of both cytochrome P450 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; α-naphthoflavone; ellipticine; furafylline
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Zamaratskaia, G.; Zlabek, V. EROD and MROD as Markers of Cytochrome P450 1A Activities in Hepatic Microsomes from Entire and Castrated Male Pigs. Sensors 2009, 9, 2134-2147.
Zamaratskaia G, Zlabek V. EROD and MROD as Markers of Cytochrome P450 1A Activities in Hepatic Microsomes from Entire and Castrated Male Pigs. Sensors. 2009; 9(3):2134-2147.
Zamaratskaia, Galia; Zlabek, Vladimir. 2009. "EROD and MROD as Markers of Cytochrome P450 1A Activities in Hepatic Microsomes from Entire and Castrated Male Pigs." Sensors 9, no. 3: 2134-2147.