Skip to main content
SpringerLink
Log in

Oxidation of barbiturates and the glucuronidation of 1-napthol in perfused rat liver and in microsomes

  • Published:
Naunyn-Schmiedeberg's Archives of Pharmacology Aims and scope Submit manuscript

Summary

An attempt has been made to relate quantitatively drug metabolism in perfused liver with that in microsomes. Therefore in both systems microsomal monooxygenase (hexobarbital as substrate) and UDP-glucuronyltransferase (1-naphthol as substrate) have been studied.

The rate of hexobarbital oxidation in perfused liver was slightly higher than in microsomes incubated with an NADPH regenerating system, indicating that the generation of reducing equivalents is not rate-limiting in livers of normal rats. The rate of phenobarbital oxidation was only about 3% of the rate of hexobarbital oxidation in perfused liver.

Microsomal UDP-glucuronyltransferase can be activated about 10-fold in vitro. The formation of naphthol glucuronide in perfused liver (determined from the appearance of glucuronide in liver tissue, perfusate and bile) corresponded well with UDP-glucuronyltransferase activity in non-activated microsomes when the intracellular UDP-glucuronate level was taken into account. This suggests that UDP-glucuronyltransferase is mostly latent within the cell.

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

  • Axelrod, J., Reichenthal, J., Brodie, B. B.: Mechanism of the potentiating action of β-diethylaminoethyl diphenylpropylacetate. J. Pharmacol. exp. Ther. 112, 49–54 (1954)

    Google Scholar 

  • Berenbom, M., Young, L.: Biochemical studies of toxic agents. Biochem. J. 49, 165–169 (1951)

    Google Scholar 

  • Bock, K. W., Fröhling, W.: UDP-glucuronyltransferase activity in isolated perfused rat liver. Naunyn-Schmiedeberg's Arch. Pharmacol. 277, 103–106 (1973)

    Google Scholar 

  • Bock, K. W., Fröhling, W., Remmer, H.: Influence of fasting and hemin on microsomal cytochromes and enzymes. Biochem. Pharmacol. 22, 1557–1564 (1973a)

    Google Scholar 

  • Bock, K. W., Fröhling, W., Remmer, H., Rexer, B.: Effects of phenobarbital and 3-methylcholanthrene on substrate specificity of rat liver microsomal UDP-glucuronyltransferase. Biochim. biophys. Acta (Amst.) 327, 46–56 (1973b)

    Google Scholar 

  • Bock, K. W., Fröhling, W., Schlote, W.: Activity and stability of microsomal mixed function oxidase and NAD glycohydrolase in isolated perfused rat liver. Naunyn-Schmiedeberg's Arch. Pharmacol. 273, 193–203 (1972)

    Google Scholar 

  • Bock, K. W., Krauss, E., Fröhling, W.: Regulation of δ-aminolevulinic acid synthetase by drugs and steroids in vivo and in isolated perfused rat liver. Europ. J. Biochem. 23, 366–371 (1971)

    Google Scholar 

  • Bock, K. W., Schwarz, L. R., White, I. N. H.: UDP-glucuronyltransferase in perfused liver and in microsomes: Influence of phenobarbital and 3-methylcholanthrene. Naunyn-Schmiedeberg's Arch. Pharmacol. 282, R 10 (1974)

    Google Scholar 

  • Bray, G. A.: A simple efficient liquid scintillator for counting aqueous solutions in a liquid scintillation counter. Analyt. Biochem. 1, 279–285 (1960)

    Google Scholar 

  • Butler, T. C.: The metabolic hydroxylation of phenobarbital. J. Pharmacol. exp. Ther. 116, 326–336 (1956)

    Google Scholar 

  • Cooper, J. R., Brodie, B. B.: The enzymatic metabolism of hexobarbital. J. Pharmacol. exp. Ther. 114, 409–417 (1955)

    Google Scholar 

  • Degkwitz, E., Ullrich, V., Staudinger, H. J., Rummel, W.: Metabolism and cytochrome P-450 binding spectra of (+)- and (−)-hexobarbital in rat liver microsomes. Hoppe-Seylers Z. physiol. Chem. 350, 547–553 (1969)

    Google Scholar 

  • Greim, H.: Synthesesteigerung und Abbauhemmung bei der Vermehrung der mikrosomalen Cytochrome P-450 und b-5 durch Phenobarbital. Naunyn-Schmiedebergs Arch. Pharmak. 266, 261–275 (1970)

    Google Scholar 

  • Hänninen, O., Puukka, R.: Effect of digitonin on UDP-glucuronyltransferase in microsomal membranes. Suom. Kemistilehti B 43, 451–456 (1970)

    Google Scholar 

  • Henderson, P. Th., Dewaide, J. H.: Metabolism of drugs in isolated rat hepatocytes. Biochem. Pharmacol. 18, 2087–2094 (1969)

    Google Scholar 

  • Hildebrandt, A., Estabrook, R. W.: Evidence for the participation of cytochrome b5 in hepatic microsomal mixed-function oxidation reactions. Arch. Biochem. Biophys. 143, 66–79 (1971)

    Google Scholar 

  • Keppler, D. O. R., Rudigier, J. F. M., Bischoff, E., Decker, K. F. A.: The trapping of uridine phosphates by d-galactosamine, d-glucosamine and 2-deoxy-d-galactose. Europ. J. Biochem. 17, 246–253 (1970)

    Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)

    Google Scholar 

  • Lucier, G. W., McDaniel, O. S., Matthews, N. B.: Microsomal rat liver UDP-glucuronyltransferase. Arch. Biochem. Biophys. 145, 520–530 (1971)

    Google Scholar 

  • Matern, S., Fröhling, W., Bock, K. W.: Albumin synthesis in isolated perfused livers from phenobarbital pretreated rats. Naunyn-Schmiedeberg's Arch. Pharmacol. 273, 242–247 (1972)

    Google Scholar 

  • Miller, L. L., Bly, C. G., Watson, M. L., Bale, W. F.: The dominant role of the liver in plasma protein synthesis. J. exp. Med. 94, 431–453 (1951)

    Google Scholar 

  • Minck, K., Schupp, R. R., Iling, H. P. A., Kahl, G. F., Netter, K. J.: Interelationship between demethylation of p-nitroanisole and conjugation of p-nitrophenol in rat liver. Naunyn-Schmiedeberg's Arch. Pharmacol. 279, 347–360 (1973)

    Google Scholar 

  • Moldeus, P. W., Cha, Y., Cinti, D. L., Schenkman, J. B.: Hepatic organelle interaction. III. Mitochondrial modification of microsomal drug metabolism. J. biol. Chem. 248, 8574–8584 (1973)

    Google Scholar 

  • Mulder, G. J.: The effect of phenobarbital on the submicrosomal distribution of uridine diphosphate glucuronyltransferase from rat liver. Biochem. J. 117, 319–324 (1970)

    Google Scholar 

  • Müller-Oerlinghausen, B., Hasselblatt, A., Jahns, R.: Vermehrte Bildung von Bilirubinglucuronid in der Leber während der Insulin- und Sulfonylharnstoff-Hypoglykämie. Naunyn-Schmiedebergs Arch. Pharmak. 260, 254–268 (1968)

    Google Scholar 

  • Remmer, H., Siegert, M.: Kumulation und Elimination von Phenobarbital. Naunyn-Schmiedebergs Arch. exp. Path. Pharmak. 243, 479–494 (1962)

    Google Scholar 

  • Remmer, H., Schenkman, J., Estabrook, R. W., Sasame, H., Gillette, J., Narasimhulu, S., Cooper, D. Y., Rosenthal, O.: Drug interaction with hepatic microsomal cytochrome. Molec. Pharmacol. 2, 187–190 (1966)

    Google Scholar 

  • Rummel, W., Brandenburger, U., Büch, H.: Versuche zur Aufklärung der Ursachen der unterschiedlichen narkotischen Wirksamkeit von (+)- und (−)-Hexobarbital. Med. Pharmacol. exp. 16, 496–504 (1967)

    Google Scholar 

  • Schenkman, J. B., Frey, I., Remmer, H., Estabrook, R. W.: Sex differences in drug metabolism by rat liver microsomes. Molec. Pharmacol. 3, 516–525 (1967)

    Google Scholar 

  • Scholz, R., Schwarz, F., Bücher, Th.: Barbiturate und energieliefernder Stoffwechsel in der hämoglobinfrei durchströmten Leber der Ratte. Z. Klin. Chem. 4, 179–189 (1966)

    Google Scholar 

  • Verity, M. A., Caper, R., Brown, W. J.: Spectrofluorometric determination of β-glucuronidase activity. Arch. Biochem. Biophys. 106, 386–393 (1964)

    Google Scholar 

  • Vessey, D. A., Zakim, D.: Regulation of microsomal enzymes by phospholipids. J. biol. Chem. 246, 4649–4656 (1971)

    Google Scholar 

  • Winsnes, A., Dutton, G. J.: Comparison between o-aminophenol glucuronidation in liver slices and homogenates from control and phenobarbital-treated Wistar and Gunn rats. Biochem. Pharmacol. 22, 1765–1771 (1973)

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Toxikologie der Universität Tübingen, Tübingen, Germany

    K. W. Bock

Authors
  1. K. W. Bock
    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

Bock, K.W. Oxidation of barbiturates and the glucuronidation of 1-napthol in perfused rat liver and in microsomes. Naunyn-Schmiedeberg's Arch. Pharmacol. 283, 319–330 (1974). https://doi.org/10.1007/BF00499191

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00499191

Key words

Search

Navigation