Skip to main content
SpringerLink
Log in

Action of methotrexate on cytochrome P-450 monooxygenases in rats

Study performed with [13C]-aminopyrine micro breath test

  • Published:
European Journal of Drug Metabolism and Pharmacokinetics Aims and scope Submit manuscript

Summary

The aim of this work was to study the action of methotrexate on the cytochrome P-450 hepatic monooxygenases and to follow the evolution of the metabolic processes with time, after oral and intra-peritoneal administration of these drugs to rats. In order to perform this study, we used a new technique the [13C]-aminopyrine breath test (ABT). At the end of the in vivo study, the rats were sacrificed and the cytochrome P-450 concentration as well as microsomal enzymatic activities of aminopyrine N-demethylase, 7-ethoxyresorufin desalkylase, 7-ethoxycoumarin desalkylase, 7-pentoxyresorufin desalkylase were determined. A histological study of the liver was also carried out. Finally, haematological and biochemical parameters were also determined. In this study, 3 series of 6 rats were used: a control group, a group receiving 70 mg/kg of methotrexate by intra-peritoneal rout, and a group of rats treated orally with 1 mg/kg of methotrexate every 2 days. For each observation, the variation of the ABT scores and of the cytochrome P-450 amounts as well as the microsomal aminopyrine N-demethylase activity were in good agreement.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Fruchart J.C. (1988): Exploration biochimique en hépatologie. In: Methais P ed. Biochimie clinique fonctionnelle. Paris. SIMEP, pp. 95–97.

    Google Scholar 

  2. Villeneuve J.P., Infante-Rivard C., Ampelas M. et al. (1986): Pronostic value of the aminopyrine breath test in cirrhotic patients. Hepatology, 6, 928–931.

    Article  CAS  PubMed  Google Scholar 

  3. Schneider J.F., Baker A.L., Haines N.W. et al. (1980): Aminopyrine N-demethylation: a prognostic test of liver function in patients with alcoholic liver disease. Gastroenterology, 79, 1145–1150.

    CAS  PubMed  Google Scholar 

  4. Irving C.S., Schoeller D.A., Nakamura K.I., Baker A.L., Klein P.D. (1982): The aminopyrine breath test as a measure of liver function. J. Lab. Clin. Med., 3, 357–369.

    Google Scholar 

  5. Bjorkman D.J., Hammond E.H., Lee R.G., Clegg D.O., Tolman K.G. (1988): Hepatic ultrastructure after methotrexate therapy for rheumatoid arthritis. Arth. Rheum., 12, 1465–1472.

    Article  Google Scholar 

  6. Reichen J., Arts B., Schafroth U., Zimmermann A., Zeltner T., Zysset T. (1987): Aminopyrine N-demethylation by rats with liver cirrhosis. Gastroenterology, 93, 719–726.

    CAS  PubMed  Google Scholar 

  7. Phillips C.A., Cera P.J., Mangan T.F., Newman E.D. (1992): Clinical liver disease in patients with rheumatoid arthritis taking methotrexate. J. Rheum., 19, 229–233.

    CAS  PubMed  Google Scholar 

  8. Guilluy R., Billion-Rey F., Brazier J.L. (1991): On-line measurement of13C enrichments in rat breath. Non-invasive method for in vivo study of drug enzymatic induction. J. Chromatogr., 562, 341–350.

    Article  CAS  PubMed  Google Scholar 

  9. Guilluy R., Billion-Rey F., Pachiaudi C. et al. (1992): On-line purification and carbon-13 isotopic analysis of carbon dioxide in breath: evaluation of on-line gas chromatography-isotope ratio mass spectrometry. Anal. Chim. Acta, 259, 193–202.

    Article  CAS  Google Scholar 

  10. Ericksson H.J., Depierre J.W., Dallner G. (1982): Preparation and properties of microsomal fractions. In: Schenkman J.B., Kûpfer D., eds. International Encyclopedia of Pharmacology & Therapeutics. New York, Pergamon Press, pp. 9–45.

    Google Scholar 

  11. Hartree E.F. (1972): Determination of protein: a modification of the Lowry method that gives a linear photometric response. Anal. Biochem., 48, 422–427.

    Article  CAS  PubMed  Google Scholar 

  12. Omura T., Sato R. (1964): The carbon-monoxide binding pigment of liver microsomes. Solubilization, purification and properties. J. Biol. Chem., 239, 2379–2385.

    CAS  PubMed  Google Scholar 

  13. Rivière J.L. (1984): Les monooxygénases hépatiques et duodénales de la caille japonaise: mise en évidence, induction et inhibition (thèse]. Paris, Université de Paris-Sud.

    Google Scholar 

  14. Rifkind A.B., Muschick H. (1983): Benoxaprofen suppression of polychlorinated biphenyl toxicity without alteration of mixed function oxidase function. Nature, 303, 524–526.

    Article  CAS  PubMed  Google Scholar 

  15. Nash T. (1953): The colorimetric estimation of formaldehyde by means of the Hantzsch reaction. J. Biochem., 239, 416–421.

    Google Scholar 

  16. Données biologiques rat OFA-SD (1991). Iffa Credo (L’Arbresle, France).

  17. Brazier J.L., Descours C., Rispal F., Billon-Rey F., Guilluy R. (1992): Inhibition enzymatique par le ditiocarb sodium (Imuthiol®). Mise en évidence par microtest respiratoire13C sur petit animal. In: Brès J., Panis G. eds. Pharmacocinétique: de la recherche à la clinique. John Libbey Eurotext©, pp. 193–198.

  18. Waxman D.J., Dannan G.A., Guengerich F.P. (1985): Development of liver microsomal P-450 isozymes in male and female rats with age. Biochemistry, 24, 4409–4417.

    Article  CAS  PubMed  Google Scholar 

  19. Guengerich F.P. ed. (1985): Mammalian cytochromes P-450. Boca Raton, Florida, pp. 16–17.

    Google Scholar 

  20. Burke D.M., Wolf R.C. (1987): Substrates and inhibitors as probes of individual forms of drug metabolising systems. In: Benford D.J., Bridges J.W., Gibson G.G., eds Drug Metabolism from Molecule to Man. pp. 219–243.

  21. Nelson D.R., Kamataki T., Waxman D.J., et al. (1993): The P450 superfamily: update on new sequences, gene mapping, accession numbers, early trivial names of enzymes, and nomenclature. DNA Cell Biol., 12, 1–51.

    Article  CAS  PubMed  Google Scholar 

  22. Plouteux G., Closon M.T. (1979): Tolérance hépatique du méthotrexate. Med. Chir. Dig., 8, 385–387.

    Google Scholar 

  23. Ishikawa M., Sasaki K., Ozaki M., Wanatabe K., Takayanagi Y., Sasaki K.I. (1991): Hepatic drug metabolizing activity in rats with carrageenan induced inflammation. J. Pharmacobiodyn., 14, 132–138.

    CAS  PubMed  Google Scholar 

  24. Endo Y., Tsuriya, S., Fujihira E. (1981): Hepatic drug metabolizing enzyme activities and antiinflammatory potency of hydrocortisone in rats with granulomatous inflammation. Res. Commun. Chem. Pathol. Pharmacol., 33, 195–206.

    CAS  PubMed  Google Scholar 

  25. Williams J.F. (1991): Cytochrome P-450 Regulation during infection, inflammation and by cytokines. J. Fla. Med. Assoc., 8, 517–519.

    Google Scholar 

  26. Mahu J.L., Feldmann G. (1984): Study of biochemical behavior of some exported and non exported hepatic proteins during acute inflammatory reaction in the rat. Enzyme, 31, 234–240.

    CAS  PubMed  Google Scholar 

  27. Breithaup H., Kuenzlen E. (1982): Pharmacokinetics of methotrexate and 7-hydroxymethotrexate following infusions of high-dose methotrexate. Cancer Treat. Rep., 66, 1733–1741.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratoire d’Etudes Analytiques et Cinétiques du Médicament, Institut des Sciences Pharmaceutiques et Biologiques de Lyon, 8 Avenue Rockefeller, 69373, Lyon, Cedex 08, France

    J. Guitton, R. Guilluy & J. L. Brazier

  2. Service d’Hématologie Pédiatrique, Hôpital Debrousse, Lyon, France

    G. Souillet

  3. Laboratoire d’Ecotoxicologie, Institut National de la Recherche Agronomique (INRA), Ecole Nationale Véterinaire de Lyon (ENVL), France

    J. L. Rivière

  4. Laboratoire d’Anatomo-pathologie de l’Institut Pasteur de Lyon, Lyon, France

    F. Gerard

Authors
  1. J. Guitton
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Souillet
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. L. Rivière
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. F. Gerard
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. R. Guilluy
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. J. L. Brazier
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Guitton, J., Souillet, G., Rivière, J.L. et al. Action of methotrexate on cytochrome P-450 monooxygenases in rats. Eur. J. Drug Metab. Pharmacokinet. 19, 119–124 (1994). https://doi.org/10.1007/BF03188832

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF03188832

Keywords

Search

Navigation