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. 1990 May 15;268(1):181–185. doi: 10.1042/bj2680181

Urobilinogen-i is a major derivative of bilirubin in bile of homozygous Gunn rats.

P Kotal 1, J Fevery 1
PMCID: PMC1131409  PMID: 2140507

Abstract

Gunn rats lack bilirubin UDP-glycosyltransferases, but diazo-negative derivatives of bilirubin have been described in their bile. In order to investigate this alternative disposal of bilirubin, crude bile samples from Gunn and Wistar rats were directly analysed by h.p.l.c. Besides bilirubin (in Gunn rats) or its glycosides (in Wistar rats), two major compounds were detected. A yellow one corresponded to the previously documented vitamin B-2 and was equally prominent in Gunn rats or Wistar-rat bile. The other compound was colourless, but on standing in contact with air it was spontaneously oxidized to a pinkish-yellow pigment. It was far more prominent in Gunn-rat bile. Analysis of bile obtained after intravenous injection of [14C]bilirubin to Gunn rats demonstrated that this compound was highly labelled. Freezing and thawing of the bile resulted in the formation of a series of diazo-negative derivatives, demonstrating that the original compound was quite labile. Spectral (adsorption and fluorescent) and chromatographic (h.p.l.c., t.l.c. and paper chromatography) analysis of the oxidized form of the labelled compound allowed its identification as urobilin-i. The colourless compound secreted in bile was urobilinogen-i. Administration of neomycin and bacitracin to Gunn rats or gut resection suppressed the biliary excretion of urobilinogen and thus confirmed its intestinal origin. Urobilinogen seems thus to represent the major bilirubin derivative present in Gunn-rat bile. Its breakdown products might represent the so-far-unidentified diazo-negative polar bilirubin derivatives. Since only a small amount of bilirubin is present in Gunn-rat bile, the urobilinogen formed in the intestinal lumen seems to be derived from bilirubin reaching the gut via routes other than the biliary one.

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Selected References

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  1. Berry C. S., Zarembo J. E., Ostrow J. D. Evidence for conversion of bilirubin to dihydroxyl derivatives in the gunn rat. Biochem Biophys Res Commun. 1972 Dec 4;49(5):1366–1375. doi: 10.1016/0006-291x(72)90617-1. [DOI] [PubMed] [Google Scholar]
  2. Blanckaert N. Analysis of bilirubin and bilirubin mono- and di-conjugates. Determination of their relative amounts in biological samples. Biochem J. 1980 Jan 1;185(1):115–128. doi: 10.1042/bj1850115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Blanckaert N., Fevery J., Heirwegh K. P., Compernolle F. Characterization of the major diazo-positive pigments in bile of homozygous Gunn rats. Biochem J. 1977 Apr 15;164(1):237–249. doi: 10.1042/bj1640237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blanckaert N., Heirwegh K. P., Zaman Z. Comparison of the biliary excretion of the four isomers of bilirubin-IX in Wistar and homozygous Gunn rats. Biochem J. 1977 Apr 15;164(1):229–236. doi: 10.1042/bj1640229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brodersen R., Bartels P. Enzymatic oxidation of bilirubin. Eur J Biochem. 1969 Oct;10(3):468–473. doi: 10.1111/j.1432-1033.1969.tb00712.x. [DOI] [PubMed] [Google Scholar]
  6. De Matteis F., Trenti T., Gibbs A. H., Greig J. B. Inducible bilirubin-degrading system in the microsomal fraction of rat liver. Mol Pharmacol. 1989 Jun;35(6):831–838. [PubMed] [Google Scholar]
  7. Fevery J., Blanckaert N., Leroy P., Michiels R., Heirwegh K. P. Analysis of bilirubins in biological fluids by extraction and thin-layer chromatography of the intact tetrapyrroles: application to bile of patients with Gilbert's syndrome, hemolysis, or cholelithiasis. Hepatology. 1983 Mar-Apr;3(2):177–183. doi: 10.1002/hep.1840030207. [DOI] [PubMed] [Google Scholar]
  8. Fevery J., Leroy P., Heirwegh K. P. Enzymic transfer of glucose and xylose from uridine diphosphate glucose and uridine diphosphate xylose to bilirubin by untreated and digitonin-activated preparations from rat liver. Biochem J. 1972 Sep;129(3):619–633. doi: 10.1042/bj1290619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HENRY R. J., JACOBS S. L., BERKMAN S. Studies on the determination of bile pigments. III. Standardization of the determination of urobilinogen as urobilinogen-aldehyde. Clin Chem. 1961 Jun;7:231–240. [PubMed] [Google Scholar]
  10. LESTER R., SCHMID R. INTESTINAL ABSORPTION OF BILE PIGMENTS. 3. THE ENTEROHEPATIC CIRCULATION OF UROBILINOGEN IN THE RAT. J Clin Invest. 1965 May;44:722–730. doi: 10.1172/JCI105185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lester R., Klein P. D. Bile pigment excretion: a comparison of the biliary excretion of bilirubin and bilirubin derivatives. J Clin Invest. 1966 Dec;45(12):1839–1846. doi: 10.1172/JCI105487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Leyten R., Vroemen J. P., Blanckaert N., Heirwegh K. P. The congenic normal R/APfd and jaundiced R/APfd-j/j rat strains: a new animal model of hereditary non-haemolytic unconjugated hyperbilirubinaemia due to defective bilirubin conjugation. Lab Anim. 1986 Oct;20(4):335–342. doi: 10.1258/002367786780808758. [DOI] [PubMed] [Google Scholar]
  13. Lloyd J. B., Weston N. T. A spectrometric study of the fluorescence detection of fecal urobilinoids. J Forensic Sci. 1982 Apr;27(2):352–365. [PubMed] [Google Scholar]
  14. Muraca M., Fevery J., Blanckaert N. Relationships between serum bilirubins and production and conjugation of bilirubin. Studies in Gilbert's syndrome, Crigler-Najjar disease, hemolytic disorders, and rat models. Gastroenterology. 1987 Feb;92(2):309–317. doi: 10.1016/0016-5085(87)90123-5. [DOI] [PubMed] [Google Scholar]
  15. OSTROW J. D., HAMMAKER L., SCHMID R. The preparation of crystalline bilirubin-C14. J Clin Invest. 1961 Aug;40:1442–1452. doi: 10.1172/JCI104375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. ROYER M., LOZZIO B. B., GORODISCH S. CHROMATOGRAPHIC SEPARATION AND PROPORTIONAL DETERMINATION OF L-, I- AND D-UROBILINS. Acta Physiol Lat Am. 1964;14:94–98. [PubMed] [Google Scholar]
  17. SCHMID R., HAMMAKER L. METABOLISM AND DISPOSITION OF C14-BILIRUBIN IN CONGENITAL NONHEMOLYTIC JAUNDICE. J Clin Invest. 1963 Nov;42:1720–1734. doi: 10.1172/JCI104858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. SCHMID R., HAMMAKER L. Metabolism and disposition of C14-bilirubin in congenital non-hemolytic jaundice. Trans Assoc Am Physicians. 1962;75:220–227. [PubMed] [Google Scholar]
  19. SCHMID R. STUDIES OF CONGENITAL NON-HEMOLYTIC JAUNDICE WITH C14-BILIRUBIN. Ann N Y Acad Sci. 1963 Dec 30;111:451–460. doi: 10.1111/j.1749-6632.1963.tb36984.x. [DOI] [PubMed] [Google Scholar]
  20. Spivak W., Yuey W. Application of a rapid and efficient h.p.l.c. method to measure bilirubin and its conjugates from native bile and in model bile systems. Potential use as a tool for kinetic reactions and as an aid in diagnosis of hepatobiliary disease. Biochem J. 1986 Feb 15;234(1):101–109. doi: 10.1042/bj2340101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stoll M. S., Gray C. H. The oxidation products of crude mesobilirubinogen. Biochem J. 1970 Apr;117(2):271–290. doi: 10.1042/bj1170271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Vermeir M., Vanstapel F., Blanckaert N. Radioassay of UDP-glucuronyltransferase-catalysed formation of bilirubin monoglucuronides and bilirubin diglucuronide in liver microsomes. Biochem J. 1984 Oct 15;223(2):455–459. doi: 10.1042/bj2230455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yokosuka O., Billing B. Enzymatic oxidation of bilirubin by intestinal mucosa. Biochim Biophys Acta. 1987 Feb 20;923(2):268–274. doi: 10.1016/0304-4165(87)90013-4. [DOI] [PubMed] [Google Scholar]

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