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Induction by Phenobarbital of Phase I and II Xenobiotic-Metabolizing Enzymes in Bovine Liver: An Overall Catalytic and Immunochemical Characterization

In cattle, phenobarbital (PB) upregulates target drug-metabolizing enzyme (DME) mRNA levels. However, few data about PB’s post-transcriptional effects are actually available. This work provides the first, and an almost complete, characterization of PB-dependent changes in DME catalytic activities in...

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Autores principales: Cantiello, Michela, Carletti, Monica, Giantin, Mery, Gardini, Giulia, Capolongo, Francesca, Cascio, Paolo, Pauletto, Marianna, Girolami, Flavia, Dacasto, Mauro, Nebbia, Carlo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8998613/
https://www.ncbi.nlm.nih.gov/pubmed/35408925
http://dx.doi.org/10.3390/ijms23073564
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author Cantiello, Michela
Carletti, Monica
Giantin, Mery
Gardini, Giulia
Capolongo, Francesca
Cascio, Paolo
Pauletto, Marianna
Girolami, Flavia
Dacasto, Mauro
Nebbia, Carlo
author_facet Cantiello, Michela
Carletti, Monica
Giantin, Mery
Gardini, Giulia
Capolongo, Francesca
Cascio, Paolo
Pauletto, Marianna
Girolami, Flavia
Dacasto, Mauro
Nebbia, Carlo
author_sort Cantiello, Michela
collection PubMed
description In cattle, phenobarbital (PB) upregulates target drug-metabolizing enzyme (DME) mRNA levels. However, few data about PB’s post-transcriptional effects are actually available. This work provides the first, and an almost complete, characterization of PB-dependent changes in DME catalytic activities in bovine liver using common probe substrates and confirmatory immunoblotting investigations. As expected, PB increased the total cytochrome P450 (CYP) content and the extent of metyrapone binding; moreover, an augmentation of protein amounts and related enzyme activities was observed for known PB targets such as CYP2B, 2C, and 3A, but also CYP2E1. However, contradictory results were obtained for CYP1A, while a decreased catalytic activity was observed for flavin-containing monooxygenases 1 and 3. The barbiturate had no effect on the chosen hydrolytic and conjugative DMEs. For the first time, we also measured the 26S proteasome activity, and the increase observed in PB-treated cattle would suggest this post-translational event might contribute to cattle DME regulation. Overall, this study increased the knowledge of cattle hepatic drug metabolism, and further confirmed the presence of species differences in DME expression and activity between cattle, humans, and rodents. This reinforced the need for an extensive characterization and understanding of comparative molecular mechanisms involved in expression, regulation, and function of DMEs.
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spelling pubmed-89986132022-04-12 Induction by Phenobarbital of Phase I and II Xenobiotic-Metabolizing Enzymes in Bovine Liver: An Overall Catalytic and Immunochemical Characterization Cantiello, Michela Carletti, Monica Giantin, Mery Gardini, Giulia Capolongo, Francesca Cascio, Paolo Pauletto, Marianna Girolami, Flavia Dacasto, Mauro Nebbia, Carlo Int J Mol Sci Article In cattle, phenobarbital (PB) upregulates target drug-metabolizing enzyme (DME) mRNA levels. However, few data about PB’s post-transcriptional effects are actually available. This work provides the first, and an almost complete, characterization of PB-dependent changes in DME catalytic activities in bovine liver using common probe substrates and confirmatory immunoblotting investigations. As expected, PB increased the total cytochrome P450 (CYP) content and the extent of metyrapone binding; moreover, an augmentation of protein amounts and related enzyme activities was observed for known PB targets such as CYP2B, 2C, and 3A, but also CYP2E1. However, contradictory results were obtained for CYP1A, while a decreased catalytic activity was observed for flavin-containing monooxygenases 1 and 3. The barbiturate had no effect on the chosen hydrolytic and conjugative DMEs. For the first time, we also measured the 26S proteasome activity, and the increase observed in PB-treated cattle would suggest this post-translational event might contribute to cattle DME regulation. Overall, this study increased the knowledge of cattle hepatic drug metabolism, and further confirmed the presence of species differences in DME expression and activity between cattle, humans, and rodents. This reinforced the need for an extensive characterization and understanding of comparative molecular mechanisms involved in expression, regulation, and function of DMEs. MDPI 2022-03-24 /pmc/articles/PMC8998613/ /pubmed/35408925 http://dx.doi.org/10.3390/ijms23073564 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Cantiello, Michela
Carletti, Monica
Giantin, Mery
Gardini, Giulia
Capolongo, Francesca
Cascio, Paolo
Pauletto, Marianna
Girolami, Flavia
Dacasto, Mauro
Nebbia, Carlo
Induction by Phenobarbital of Phase I and II Xenobiotic-Metabolizing Enzymes in Bovine Liver: An Overall Catalytic and Immunochemical Characterization
title Induction by Phenobarbital of Phase I and II Xenobiotic-Metabolizing Enzymes in Bovine Liver: An Overall Catalytic and Immunochemical Characterization
title_full Induction by Phenobarbital of Phase I and II Xenobiotic-Metabolizing Enzymes in Bovine Liver: An Overall Catalytic and Immunochemical Characterization
title_fullStr Induction by Phenobarbital of Phase I and II Xenobiotic-Metabolizing Enzymes in Bovine Liver: An Overall Catalytic and Immunochemical Characterization
title_full_unstemmed Induction by Phenobarbital of Phase I and II Xenobiotic-Metabolizing Enzymes in Bovine Liver: An Overall Catalytic and Immunochemical Characterization
title_short Induction by Phenobarbital of Phase I and II Xenobiotic-Metabolizing Enzymes in Bovine Liver: An Overall Catalytic and Immunochemical Characterization
title_sort induction by phenobarbital of phase i and ii xenobiotic-metabolizing enzymes in bovine liver: an overall catalytic and immunochemical characterization
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8998613/
https://www.ncbi.nlm.nih.gov/pubmed/35408925
http://dx.doi.org/10.3390/ijms23073564
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