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. 1989 Apr;83(4):1211–1216. doi: 10.1172/JCI114003

Downregulation of the male-specific hepatic microsomal steroid 16 alpha-hydroxylase, cytochrome P-450UT-A, in rats with portal bypass. Relevance to estradiol accumulation and impaired drug metabolism in hepatic cirrhosis.

E Cantrill 1, M Murray 1, I Mehta 1, G C Farrell 1
PMCID: PMC303809  PMID: 2703529

Abstract

Elevated serum estradiol concentrations and specific changes in the biliary excretion of some androstenedione metabolites have been reported in male rats with portal bypass produced by portal vein ligation (PVL). In this study, the hypothesis that male-specific forms of cytochrome P-450 are altered after PVL was tested by measuring microsomal steroid hydroxylase activities. Consistent with earlier findings in the intact animal, androstenedione 16 alpha-hydroxylase activity was reduced after PVL to 44% of control (P less than 0.05). Other pathways of androstenedione hydroxylation, and total estrogen formation (after androstenedione aromatization) were unchanged. Although total estrogen formation was not different, a sevenfold greater proportion of estradiol was produced in PVL rat microsomes. Additional experiments revealed that PVL selectively reduced the rate of microsomal estradiol 16 alpha-hydroxylation (to 56% of control, P less than 0.02). Levels of cytochrome P-450UT-A, the microsomal steroid 16 alpha-hydroxylase, were lower after PVL (56% of control, P less than 0.05), so that the present observations are consistent with the earlier suggestion that portal bypass is associated with the selective downregulation of this enzyme. Since downregulation of cytochrome P-450UT-A also occurs in experimental hepatic cirrhosis, portal hypertension may well contribute significantly to altered drug metabolism in liver disease. Impaired hepatic elimination of androstenedione by hydroxylation may indirectly enhance extrahepatic aromatization of the androgen. The decreased activity of hepatic estradiol 16 alpha-hydroxylation after PVL would enhance the accumulation of estradiol, the biologically more potent estrogen.

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Selected References

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  1. Blomquist C. H., Kotts C. E., Hakanson E. Y. Microsomal 17beta-hydroxysteroid dehydrogenase of guinea pig liver: detergent solubilization and a comparison of kinetic and stability properties of bound and solubilized forms. J Steroid Biochem. 1977 Mar;8(3):193–198. doi: 10.1016/0022-4731(77)90050-4. [DOI] [PubMed] [Google Scholar]
  2. Cameron D. P., Burger H. G., Catt K. J., Gordon E., Watts J. M. Metabolic clearance of human growth hormone in patients with hepatic and renal failure, and in the isolated perfused pig liver. Metabolism. 1972 Oct;21(10):895–904. doi: 10.1016/0026-0495(72)90024-8. [DOI] [PubMed] [Google Scholar]
  3. Cheng K. C., Schenkman J. B. Metabolism of progesterone and estradiol by microsomes and purified cytochrome P-450 RLM3 and RLM5. Drug Metab Dispos. 1984 Mar-Apr;12(2):222–234. [PubMed] [Google Scholar]
  4. Conney A. H. Pharmacological implications of microsomal enzyme induction. Pharmacol Rev. 1967 Sep;19(3):317–366. [PubMed] [Google Scholar]
  5. Dannan G. A., Guengerich F. P., Waxman D. J. Hormonal regulation of rat liver microsomal enzymes. Role of gonadal steroids in programming, maintenance, and suppression of delta 4-steroid 5 alpha-reductase, flavin-containing monooxygenase, and sex-specific cytochromes P-450. J Biol Chem. 1986 Aug 15;261(23):10728–10735. [PubMed] [Google Scholar]
  6. Dannan G. A., Porubek D. J., Nelson S. D., Waxman D. J., Guengerich F. P. 17 beta-estradiol 2- and 4-hydroxylation catalyzed by rat hepatic cytochrome P-450: roles of individual forms, inductive effects, developmental patterns, and alterations by gonadectomy and hormone replacement. Endocrinology. 1986 May;118(5):1952–1960. doi: 10.1210/endo-118-5-1952. [DOI] [PubMed] [Google Scholar]
  7. Einarsson K., Gustafsson J. A., Stenberg A. Neonatal imprinting of liver microsomal hydroxylation and reduction of steroids. J Biol Chem. 1973 Jul 25;248(14):4987–4997. [PubMed] [Google Scholar]
  8. Farrell G. C., Koltai A., Murray M. Source of raised serum estrogens in male rats with portal bypass. J Clin Invest. 1988 Jan;81(1):221–228. doi: 10.1172/JCI113299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Farrell G. C., Koltai A., Zaluzny L., Murray M. Effects of portal vein ligation on sex hormone metabolism in male rats: relationship to lowered hepatic cytochrome P450 levels. Gastroenterology. 1986 Feb;90(2):299–305. doi: 10.1016/0016-5085(86)90924-8. [DOI] [PubMed] [Google Scholar]
  10. Farrell G. C., Zaluzny L. Portal vein ligation selectively lowers hepatic cytochrome P450 levels in rats. Gastroenterology. 1983 Aug;85(2):275–282. [PubMed] [Google Scholar]
  11. Fishman J., Bradlow H. L., Schneider J., Anderson K. E., Kappas A. Radiometric analysis of biological oxidations in man: sex differences in estradiol metabolism. Proc Natl Acad Sci U S A. 1980 Aug;77(8):4957–4960. doi: 10.1073/pnas.77.8.4957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Guengerich F. P., Dannan G. A., Wright S. T., Martin M. V., Kaminsky L. S. Purification and characterization of liver microsomal cytochromes p-450: electrophoretic, spectral, catalytic, and immunochemical properties and inducibility of eight isozymes isolated from rats treated with phenobarbital or beta-naphthoflavone. Biochemistry. 1982 Nov 9;21(23):6019–6030. doi: 10.1021/bi00266a045. [DOI] [PubMed] [Google Scholar]
  13. Guengerich F. P. Isolation and purification of cytochrome P-450, and the existence of multiple forms. Pharmacol Ther. 1979;6(1):99–121. doi: 10.1016/0163-7258(79)90057-3. [DOI] [PubMed] [Google Scholar]
  14. Jellinck P. H., Quail J. A., Crowley C. A. Normal and recombinant human growth hormone administered by constant infusion feminize catechol estrogen formation by rat liver microsomes. Endocrinology. 1985 Dec;117(6):2274–2278. doi: 10.1210/endo-117-6-2274. [DOI] [PubMed] [Google Scholar]
  15. Kamataki T., Maeda K., Yamazoe Y., Nagai T., Kato R. Sex difference of cytochrome P-450 in the rat: purification, characterization, and quantitation of constitutive forms of cytochrome P-450 from liver microsomes of male and female rats. Arch Biochem Biophys. 1983 Sep;225(2):758–770. doi: 10.1016/0003-9861(83)90087-5. [DOI] [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  18. Lauterburg B. H., Sautter V., Preisig R., Bircher J. Hepatic functional deterioration after portacaval shunt in the rat. Effects on sulfobromophthalein transport-maximum, indocyanine green clearance and galactose elimination capacity. Gastroenterology. 1976 Aug;71(2):221–227. [PubMed] [Google Scholar]
  19. Mason D. Y., Sammons R. Alkaline phosphatase and peroxidase for double immunoenzymatic labelling of cellular constituents. J Clin Pathol. 1978 May;31(5):454–460. doi: 10.1136/jcp.31.5.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Milewich L., Garcia R. L., Gerrity L. W. 17 beta-Hydroxysteroid oxidoreductase: a ubiquitous enzyme. Interconversion of estrone and estradiol-17 beta in BALB/c mouse tissues. Metabolism. 1985 Oct;34(10):938–944. doi: 10.1016/0026-0495(85)90142-8. [DOI] [PubMed] [Google Scholar]
  21. Mode A., Gustafsson J. A., Jansson J. O., Edén S., Isaksson O. Association between plasma level of growth hormone and sex differentiation of hepatic steroid metabolism in the rat. Endocrinology. 1982 Nov;111(5):1692–1697. doi: 10.1210/endo-111-5-1692. [DOI] [PubMed] [Google Scholar]
  22. Morgan E. T., MacGeoch C., Gustafsson J. A. Hormonal and developmental regulation of expression of the hepatic microsomal steroid 16 alpha-hydroxylase cytochrome P-450 apoprotein in the rat. J Biol Chem. 1985 Oct 5;260(22):11895–11898. [PubMed] [Google Scholar]
  23. Murray M., Cantrill E., Farrell G. C. Hepatic microsomal metabolism of androst-4-ene-3,17-dione: relative importance of ring hydroxylation and aromatization in control and induced rat liver. J Steroid Biochem. 1988 Feb;29(2):233–237. doi: 10.1016/0022-4731(88)90271-3. [DOI] [PubMed] [Google Scholar]
  24. Murray M. Complexation of cytochrome P-450 isozymes in hepatic microsomes from SKF 525-A-induced rats. Arch Biochem Biophys. 1988 May 1;262(2):381–388. doi: 10.1016/0003-9861(88)90202-0. [DOI] [PubMed] [Google Scholar]
  25. Murray M., Zaluzny L., Dannan G. A., Guengerich F. P., Farrell G. C. Altered regulation of cytochrome P-450 enzymes in choline-deficient cirrhotic male rat liver: impaired regulation and activity of the male-specific androst-4-ene-3,17-dione 16 alpha-hydroxylase, cytochrome P-450UT-A, in hepatic cirrhosis. Mol Pharmacol. 1987 Jan;31(1):117–121. [PubMed] [Google Scholar]
  26. Murray M., Zaluzny L., Farrell G. C. Drug metabolism in cirrhosis. Selective changes in cytochrome P-450 isozymes in the choline-deficient rat model. Biochem Pharmacol. 1986 Jun 1;35(11):1817–1824. doi: 10.1016/0006-2952(86)90298-4. [DOI] [PubMed] [Google Scholar]
  27. Murray M., Zaluzny L., Farrell G. C. Impaired androgen 16 alpha-hydroxylation in hepatic microsomes from carbon tetrachloride-cirrhotic male rats. Gastroenterology. 1987 Jul;93(1):141–147. doi: 10.1016/0016-5085(87)90326-x. [DOI] [PubMed] [Google Scholar]
  28. Murray M., Zaluzny L., Farrell G. C. Selective reactivation of steroid hydroxylases following dissociation of the isosafrole metabolite complex with rat hepatic cytochrome P-450. Arch Biochem Biophys. 1986 Dec;251(2):471–478. doi: 10.1016/0003-9861(86)90354-1. [DOI] [PubMed] [Google Scholar]
  29. Nerland D. E., Mannering G. J. Species, sex, and developmental differences in the O- and N-dealkylation of ethylmorphine by hepatic microsomes. Drug Metab Dispos. 1978 Mar-Apr;6(2):150–153. [PubMed] [Google Scholar]
  30. Schimpff R. M., Lebrec D., Dannadieu M., Repellin A. M. Serum somatomedin activity measured as sulphation factor in peripheral, hepatic and renal veins of patients with alcoholic cirrhosis. Acta Endocrinol (Copenh) 1978 Aug;88(4):729–736. doi: 10.1530/acta.0.0880729. [DOI] [PubMed] [Google Scholar]
  31. Smuk M., Schwers J. Aromatization of androstenedione by human adult liver in vitro. J Clin Endocrinol Metab. 1977 Nov;45(5):1009–1012. doi: 10.1210/jcem-45-5-1009. [DOI] [PubMed] [Google Scholar]
  32. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Van Thiel D. H., Gavaler J. S., Schade R. R. Liver disease and the hypothalamic pituitary gonadal axis. Semin Liver Dis. 1985 Feb;5(1):35–45. doi: 10.1055/s-2008-1041756. [DOI] [PubMed] [Google Scholar]
  34. Van Thiel D. H., Gavaler J. S., Wight C., Smith W. I., Jr, Abuid J. Thyrotropin-releasing hormone (TRH)-induced growth hormone (hGH) responses in cirrhotic men. Gastroenterology. 1978 Jul;75(1):66–70. [PubMed] [Google Scholar]
  35. Van Thiel D. H., Lester R., Vaitukaitis J. Evidence for a defect in pituitary secretion of luteinizing hormone in chronic alcoholic men. J Clin Endocrinol Metab. 1978 Sep;47(3):499–507. doi: 10.1210/jcem-47-3-499. [DOI] [PubMed] [Google Scholar]
  36. Waxman D. J., Dannan G. A., Guengerich F. P. Regulation of rat hepatic cytochrome P-450: age-dependent expression, hormonal imprinting, and xenobiotic inducibility of sex-specific isoenzymes. Biochemistry. 1985 Jul 30;24(16):4409–4417. doi: 10.1021/bi00337a023. [DOI] [PubMed] [Google Scholar]
  37. Waxman D. J. Interactions of hepatic cytochromes P-450 with steroid hormones. Regioselectivity and stereospecificity of steroid metabolism and hormonal regulation of rat P-450 enzyme expression. Biochem Pharmacol. 1988 Jan 1;37(1):71–84. doi: 10.1016/0006-2952(88)90756-3. [DOI] [PubMed] [Google Scholar]
  38. Waxman D. J., Ko A., Walsh C. Regioselectivity and stereoselectivity of androgen hydroxylations catalyzed by cytochrome P-450 isozymes purified from phenobarbital-induced rat liver. J Biol Chem. 1983 Oct 10;258(19):11937–11947. [PubMed] [Google Scholar]
  39. Waxman D. J., Lapenson D. P., Park S. S., Attisano C., Gelboin H. V. Monoclonal antibodies inhibitory to rat hepatic cytochromes P-450: P-450 form specificities and use as probes for cytochrome P-450-dependent steroid hydroxylations. Mol Pharmacol. 1987 Nov;32(5):615–624. [PubMed] [Google Scholar]
  40. Waxman D. J. Rat hepatic cytochrome P-450 isoenzyme 2c. Identification as a male-specific, developmentally induced steroid 16 alpha-hydroxylase and comparison to a female-specific cytochrome P-450 isoenzyme. J Biol Chem. 1984 Dec 25;259(24):15481–15490. [PubMed] [Google Scholar]
  41. Wood A. W., Ryan D. E., Thomas P. E., Levin W. Regio- and stereoselective metabolism of two C19 steroids by five highly purified and reconstituted rat hepatic cytochrome P-450 isozymes. J Biol Chem. 1983 Jul 25;258(14):8839–8847. [PubMed] [Google Scholar]
  42. Wu A., Grant D. B., Hambley J., Levi A. J. Reduced serum somatomedin activity in patients with chronic liver disease. Clin Sci Mol Med. 1974 Oct;47(4):359–366. doi: 10.1042/cs0470359. [DOI] [PubMed] [Google Scholar]
  43. YATES F. E., HERBST A. L., URQUHART J. Sex difference in rate of ring A reduction of delta 4-3-keto-steroids in vitro by rat liver. Endocrinology. 1958 Dec;63(6):887–902. doi: 10.1210/endo-63-6-887. [DOI] [PubMed] [Google Scholar]
  44. Yamazoe Y., Shimada M., Murayama N., Kato R. Suppression of levels of phenobarbital-inducible rat liver cytochrome P-450 by pituitary hormone. J Biol Chem. 1987 May 25;262(15):7423–7428. [PubMed] [Google Scholar]

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