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Gene and protein expression and cellular localisation of cytochrome P450 enzymes of the 1A, 2A, 2C, 2D and 2E subfamilies in equine intestine and liver.
Abstract: Among the cytochrome P450 enzymes (CYP), families 1-3 constitute almost half of total CYPs in mammals and play a central role in metabolism of a wide range of pharmaceuticals. This study investigated gene and protein expression and cellular localisation of CYP1A, CYP2A, CYP2C, CYP2D and CYP2E in equine intestine and liver. Real-time polymerase chain reaction (RT-PCR) was used to analyse gene expression, western blot to examine protein expression and immunohistochemical analyses to investigate cellular localisation. Results: CYP1A and CYP2C were the CYPs with the highest gene expression in the intestine and also showed considerable gene expression in the liver. CYP2E and CYP2A showed the highest gene expression in the liver. CYP2E showed moderate intestinal gene expression, whereas that of CYP2A was very low or undetectable. For CYP2D, rather low gene expression levels were found in both intestine and the liver. In the intestine, CYP gene expression levels, except for CYP2E, exhibited patterns resembling those of the proteins, indicating that intestinal protein expression of these CYPs is regulated at the transcriptional level. For CYP2E, the results showed that the intestinal gene expression did not correlate to any visible protein expression, indicating that intestinal protein expression of this CYP is regulated at the post-transcriptional level. Immunostaining of intestine tissue samples showed preferential CYP staining in enterocytes at the tips of intestinal villi in the small intestine. In the liver, all CYPs showed preferential localisation in the centrilobular hepatocytes. Conclusions: Overall, different gene expression profiles were displayed by the CYPs examined in equine intestine and liver. The CYPs present in the intestine may act in concert with those in the liver to affect the oral bioavailability and therapeutic efficiency of substrate drugs. In addition, they may play a role in first-pass metabolism of feed constituents and of herbal supplements used in equine practice.
Publication Date: 2014-10-08 PubMed ID: 25288196PubMed Central: PMC4192735DOI: 10.1186/s13028-014-0069-8Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article investigates the expression and location of various cytochrome P450 enzymes – particularly CYP1A, CYP2A, CYP2C, CYP2D, and CYP2E – in the liver and intestines of horses. It also analyzes the impact of these enzymes on the metabolism of pharmaceuticals and other substances.
Research Design and Methods
- The study utilised real-time polymerase chain reaction (RT-PCR) to examine gene expression, western blot for protein expression, and immunohistochemical analyses to determine cellular localisation.
- The concentrations of CYP1A, CYP2A, CYP2C, CYP2D and CYP2E in the equine intestine and liver were particularly investigated.
Results and Findings
- CYP1A and CYP2C had the highest gene expression in the intestine and significant gene expression in the liver.
- CYP2E and CYP2A had the highest gene expression in the liver.
- Within the intestine, CYP2E had moderate gene expression while CYP2A’s gene expression was minimal or unobservable.
- CYP2D gene expression in both the liver and intestines was relatively low.
- Excluding CYP2E, gene expression levels for CYP in the intestines resembled patterns seen in protein expression levels, suggesting protein expression of these CYPs in the intestine is regulated at a transcriptional level.
- For CYP2E, there seemed to be no correlation between gene and protein expression in the intestines, suggesting a post-transcriptional regulation.
- Cellular localisation was examined through immunostaining, which revealed a preferential localisation of CYP in enterocytes at the tips of intestinal villi in the small intestine, and in the liver’s centrilobular hepatocytes.
Conclusions
- The research found different gene expression profiles for the examined CYPs in the equine intestine and liver.
- The study suggests that the various CYPs present in the intestines can work together with those in the liver to influence the bioavailability and therapeutic efficiency of substrate drugs.
- Lastly, it implies that these CYPs may be part of the first-pass metabolism of feed constituents and herbal supplements used in equine practice.
Cite This Article
APA
Tydén E, Tjälve H, Larsson P.
(2014).
Gene and protein expression and cellular localisation of cytochrome P450 enzymes of the 1A, 2A, 2C, 2D and 2E subfamilies in equine intestine and liver.
Acta Vet Scand, 56(1), 69.
https://doi.org/10.1186/s13028-014-0069-8 Publication
Researcher Affiliations
- Department of Biomedical Sciences and Veterinary Public Health, Division of Pathology, Pharmacology and Toxicology, Swedish University of Agricultural Sciences, Uppsala, S-750 07, Sweden. eva.tyden@slu.se.
MeSH Terms
- Animals
- Cytochrome P-450 Enzyme System / genetics
- Cytochrome P-450 Enzyme System / metabolism
- Female
- Gene Expression
- Horses / genetics
- Horses / metabolism
- Intestinal Mucosa / metabolism
- Liver / metabolism
- Male
- Organ Specificity
- Real-Time Polymerase Chain Reaction / veterinary
- Reverse Transcriptase Polymerase Chain Reaction / veterinary
References
This article includes 51 references
Citations
This article has been cited 4 times.- Graves JP, Bradbury JA, Gruzdev A, Li H, Duval C, Lih FB, Edin ML, Zeldin DC. Expression of Cyp2c/Cyp2j subfamily members and oxylipin levels during LPS-induced inflammation and resolution in mice. FASEB J 2019 Dec;33(12):14784-14797.
- Zhou L, Cui M, Zhao L, Wang D, Tang T, Wang W, Wang S, Huang H, Qiu X. Potential Metabolic Drug-Drug Interaction of Citrus aurantium L. (Rutaceae) Evaluating by Its Effect on 3 CYP450. Front Pharmacol 2018;9:895.
- Nemeth Z, Eros K, Munkacsy G, Koller A. Differential Gene and Protein Expressions Responsible for Vasomotor Signaling Provide Mechanistic Bases for the Opposite Flow-Induced Responses of Pre- and Post-Circle of Willis Arteries. Life (Basel) 2025 May 26;15(6).
- Knauer JF, Schulz C, Zemella A, Wüstenhagen DA, Walter RM, Küpper JH, Kubick S. Synthesis of mono Cytochrome P450 in a modified CHO-CPR cell-free protein production platform. Sci Rep 2024 Jan 13;14(1):1271.
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