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Research ArticleArticle

Effect of Breed upon Cytochromes P450 and Phase II Enzyme Expression in Cattle Liver

Mery Giantin, Monica Carletti, Francesca Capolongo, Sara Pegolo, Rosa Maria Lopparelli, Federica Gusson, Carlo Nebbia, Michela Cantiello, Pascal Martin, Thierry Pineau and Mauro Dacasto
Drug Metabolism and Disposition May 2008, 36 (5) 885-893; DOI: https://doi.org/10.1124/dmd.107.019042
Mery Giantin
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Monica Carletti
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Francesca Capolongo
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Sara Pegolo
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Rosa Maria Lopparelli
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Federica Gusson
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Carlo Nebbia
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Michela Cantiello
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Pascal Martin
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Thierry Pineau
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Mauro Dacasto
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Abstract

Cattle represent an important source of animal-derived food-products; nonetheless, our knowledge about the expression of drug-metabolizing enzymes (DMEs) in present and other food-producing animals still remains superficial, despite the obvious toxicological consequences. Breed represents an internal factor that modulates DME expression and catalytic activity. In the present work, the effect of breed upon relevant phase I and phase II DMEs was investigated at the pretranscriptional and post-translational levels in male Charolais (CH), Piedmontese (PM) and Blonde d'Aquitaine (BA) cattle. Because specific substrates for cattle have not yet been identified, the breed effect upon specific cytochrome P450 (P450), UDP-glucuronosyltransferase (UGT), or glutathione S-transferase (GST) DMEs, in terms of catalytic activity, was determined by using human marker substrates. Among P450s, benzphetamine N-demethylase, 16β-, 6β-, and 2β-testosterone hydroxylase, aniline and p-nitrophenol hydroxylase, and α-naphthol and p-nitrophenol UGT activities were significantly higher in CH; in contrast, lower levels of CYP1A1-, CYP1A2-, CYP2B6-, CYP2C9-, CYP2C18-, CYP3A4-, and UGT1A1-like mRNAs were noticed, with CH < PM ≤ BA as a trend. CYP2B and CYP3A mRNA results were confirmed with immunoblotting, too. As regards conjugative DMEs, UGT1A6-like mRNA levels were consistent with respective catalytic activities. Both 1-chloro-2,4-dinitrobenzene and 3,4-dichloronitrobenzene GST activities were higher in BA, and these results agreed with GSTA1-, GSTM1-, and GSTP1-like mRNA amounts. Correlation analysis between catalytic activities and mRNAs showed either significant or uneven results, depending on the substrate. These findings confirm previous data obtained in laboratory species; however, further studies are required to ascribe this behavior to pretranscriptional or post-translational phenomena.

Footnotes

  • This study was supported by the Università degli Studi di Padova (Grants 60A08-8213/05, Studi sull'espressione degli enzimi farmaco-metabolizzanti nel fegato di bovini appartenenti a razze diverse; 60A08-7722/06, Studi sull'espressione di enzimi coniugativi nel fegato di bovini appartenenti a razze diverse; 60A08-3508/07, Valutazione dei profili di espressione di geni codificanti per gli enzimi farmaco-metabolizzanti nel fegato di bovini appartenenti a razze diverse to M.D.), by the Ministero Università e Ricerca Scientifica e Tecnologica and Università Italo-Francese (Valutazione degli effetti dei trattamenti illeciti sugli enzimi farmaco-metabolizzanti del bovino mediante tecniche di real-time PCR) (to M.D. and T.P.), by the Università degli Studi di Torino (2-year postdoctoral fellowship to M.C.), and by the Università degli Studi di Padova (Ph.D. fellowship to M.G.).

  • Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.

  • doi:10.1124/dmd.107.019042.

  • ABBREVIATIONS: DME, drug-metabolizing enzyme; P450, cytochrome P450; PM, Piedmontese; CH, Charolais; BA, Blonde d'Aquitaine; Q RT-PCR, quantitative real-time polymerase chain reaction; HPLC, high-performance liquid chromatography; PBS, phosphate-buffered saline; TST, testosterone; 2βOH-T, 2β-testosterone hydroxylase; 6βOH-T, 6β-testosterone hydroxylase; 16βOH-T, 16β-testosterone hydroxylase; EROD, ethoxyresorufin O-deethylase; ETR, ethoxyresorufin; UGT, UDP-glucuronosyltransferase; GST, glutathione S-transferase; CDNB, 1-chloro-2,4-dinitrobenzene; DCNB, 3,4-dichloronitrobenzene; GSH, glutathione; ANOVA, analysis of variance; BENZDEM, benzphetamine N-demethylase; TBT4-OH, tolbutamide 4-hydroxylase; Aniline-OH, aniline hydroxylase; pNP-OH, p-nitrophenol hydroxylase; ETDEM, ethylmorphine N-demethylase; ERDEM, erythromycin N-demethylase; TAODEM, triacetyloleandomycin N-demethylase; a.u., arbitrary unit(s).

    • Received September 27, 2007.
    • Accepted February 6, 2008.
  • The American Society for Pharmacology and Experimental Therapeutics
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Drug Metabolism and Disposition: 36 (5)
Drug Metabolism and Disposition
Vol. 36, Issue 5
1 May 2008
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Research ArticleArticle

Effect of Breed upon Cytochromes P450 and Phase II Enzyme Expression in Cattle Liver

Mery Giantin, Monica Carletti, Francesca Capolongo, Sara Pegolo, Rosa Maria Lopparelli, Federica Gusson, Carlo Nebbia, Michela Cantiello, Pascal Martin, Thierry Pineau and Mauro Dacasto
Drug Metabolism and Disposition May 1, 2008, 36 (5) 885-893; DOI: https://doi.org/10.1124/dmd.107.019042

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Research ArticleArticle

Effect of Breed upon Cytochromes P450 and Phase II Enzyme Expression in Cattle Liver

Mery Giantin, Monica Carletti, Francesca Capolongo, Sara Pegolo, Rosa Maria Lopparelli, Federica Gusson, Carlo Nebbia, Michela Cantiello, Pascal Martin, Thierry Pineau and Mauro Dacasto
Drug Metabolism and Disposition May 1, 2008, 36 (5) 885-893; DOI: https://doi.org/10.1124/dmd.107.019042
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