Advertisement

Journal of Gastroenterology

, Volume 31, Issue 6, pp 828–835 | Cite as

Deconjugation of bilirubin accelerates coprecipitation of cholesterol, fatty acids, and mucin in human bile—In vitro study

  • Hidetaka Higashijima
  • Hitoshi Ichimiya
  • Toru Nakano
  • Hiroyuki Yamashita
  • Shoji Kuroki
  • Hiroshi Satoh
  • Kazuo Chijiiwa
  • Masao Tanaka
Liver, Pancreas, and Biliary Tract

Abstract

To examine the initial step of brown pigment gallstone formation, sterile human gallbladder bile samples were incubated with or without β-glucuronidase in vitro. Enhanced bilirubin deconjugation achieved by adding β-glucuronidase significantly accelerated the formation of a precipitate that contained bilirubin (28.2±3.8% of dry weight), cholesterol (14.3±5.2%), free fatty acids (12.0±1.3%), and glycoprotein (10.0±6.7%). Both the composition and scanning electron microscopic appearance of the precipitate were similar to these features in brown pigment gallstones. The cholesterol saturation index and nucleation time in the supernatant did not change with various incubation periods. The weight ratios of bilirubin to cholesterol in the precipitates correlated with those in bile (r=0.76;P=0.017). Gel chromatography of the precipitate showed high molecular weight glycoprotein to be the major constituent. Bilirubin, cholesterol, fatty acids, and mucin were found to coprecipitate in accordance with bilirubin deconjugation, which process may play an important role in an early stage of the formation of brown pigment gallstones.

Key words

Brown pigment gallstone calcium bilirubinate bilirubin cholesterol free fatty acids β-glucuronidase 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Soloway RD, Trotman BW, Ostrow JD. Progress in gastroenterology. Pigment gallstone. Gastroenterology 1977;72:167–181.PubMedGoogle Scholar
  2. 2.
    Nakayama F. Quantitative microanalysis of gallstones. J Lab Clin Med 1968;72:602–611.PubMedGoogle Scholar
  3. 3.
    Tabata M, Nakayama F. Bacteria and gallstones. Etiological significance. Dig Dis Sci 1981;26:218–224.CrossRefPubMedGoogle Scholar
  4. 4.
    Cetta FM. Bile infection documented as initial event in the pathogenesis of brown pigment biliary stone. Gastroenterology 1986;6:482–489.Google Scholar
  5. 5.
    Maki T. Pathogenesis of calcium bilirubinate gallstone: Role ofE. coli, β-glucuronidase and coagulation by inorganic ions, polyelectrolytes and agitation. Ann Surg 1964;164:90–100.Google Scholar
  6. 6.
    Junipor K, Burson EN. Biliary tract studies. The significance of biliary crystals. Gastroenterology 1957;32:175–211.Google Scholar
  7. 7.
    Van Erpecum KJ, Van Berge Henegouwen GP, Stoelwinder B, et al. Cholesterol and pigment gallstone dis-ease: Comparison of three bile tests for differentiation bet-ween the two stone types. Scand J Gastroenterol 1988;23:948–954.PubMedGoogle Scholar
  8. 8.
    Nakano T, Yanagisawa J, Nakayama F. Phospholipase activity in human bile. Hepatology 1988;8:1560–1564.PubMedGoogle Scholar
  9. 9.
    Chijiiwa K, Hirota I, Noshiro H. High vesicular cholesterol and protein in bile are associated with formation of cholesterol but not pigment gallstones. Dig Dis Sci 1993;38:161–166.CrossRefPubMedGoogle Scholar
  10. 10.
    Chijiiwa K, Nakayama F. Simultaneous microanalysis of bile acids and cholesterol in bile by glass capillary column gas chromatography. J Chromatogr 1988;431:17–25.PubMedGoogle Scholar
  11. 11.
    Talalay P. Enzymatic analysis of steroid hormone. Methods Biochem Anal 1960;8:119–143.PubMedGoogle Scholar
  12. 12.
    Bartlett GR. Phosphorus assay in column chromatography. J Biol Chem 1959;234:466–468.PubMedGoogle Scholar
  13. 13.
    Kuroki S, Cohen BI, Carey MC, et al. Rapid cholesterol saturation of bile sample. J Lipid Res 1986;27:442–446.PubMedGoogle Scholar
  14. 14.
    Carey MC. Critical tables for calculating the cholesterol saturation of native bile. J Lipid Res 1978;19:945–955.PubMedGoogle Scholar
  15. 15.
    Dole VP, Meinartz H. Microdetermination of long-chain fatty acids in plasma and tissues. J Biol Chem 1960;235:2595–2599.PubMedGoogle Scholar
  16. 16.
    Michaelsson M. Bilirubin determinations in serum and urine. Studies on diazo methods and a new copper azopigment method. Scand J Clin Lab Invest 1961;13(Suppl 56):1–80.Google Scholar
  17. 17.
    Spivak W, Yuey W. Application of a rapid and efficient HPLC method to measure bilirubin and its conjugates from native bile and in model bile system. Biochem J 1986;234:101–109.PubMedGoogle Scholar
  18. 18.
    Malloy HT, Evelyn KA. The determination of bilirubin with a photoelectric colorimeter. J Biol Chem 1937;119:481–490.Google Scholar
  19. 19.
    LaMont JT, Ventola AS, Trotman BW, et al. Mucin glycoprotein content of human pigment gallstone. Hepatology 1983;3:377–382.PubMedGoogle Scholar
  20. 20.
    Mantle M, Allen A. A colorimetric assay for glycoproteins based on the periodic acid/Schiff stain. Biochem Soc Trans 1978;6:607–609.PubMedGoogle Scholar
  21. 21.
    Messing B, Boris C, Kunstringen F, et al. Does total parenteral nutrition induce gallbladder sludge formation and lithiasis? Gastroenterology 1983;84:1012–1019.PubMedGoogle Scholar
  22. 22.
    Lee SP, Nicholls JF. Nature and composition of biliary sludge. Gastroenterology 1986;90:677–686.PubMedGoogle Scholar
  23. 23.
    Yamashita N, Nakayama F. Composition of intrahepatic calculi. Etiological significance. Dig Dis Sci 1988;33:449–453.CrossRefPubMedGoogle Scholar
  24. 24.
    Oyabu H, Tabata M, Nakayama F. Nonbacterial transformation of bilirubin in bile. Dig Dis Sci 1987;32:809–816.CrossRefPubMedGoogle Scholar
  25. 25.
    Ho KJ, Hsu SC, Chen JS, et al. Human biliary β-glucuronidase. Correlation of its activity with deconjugation of bilirubin in the bile. Eur J Clin Invest 1986;16:361–367.PubMedGoogle Scholar
  26. 26.
    Sjödahl R, Tagesson C. Formation and inhibition of lysolecithin in human gallbladder bile. Acta Chir Scand 1976;142:395–399.PubMedGoogle Scholar
  27. 27.
    Smith BF, LaMont JT. Bovine gallbladder mucin binds bilirubin in vitro. Gastroenterology 1983;85:707–712.PubMedGoogle Scholar
  28. 28.
    Smith BF, LaMont BF. Hydrophobic binding properties of bovine gallbladder mucin. J Biol Chem 1984;259:12170–12177.PubMedGoogle Scholar
  29. 29.
    Gong D, Turner B, Bhaskar KR, et al. Lipid binding to gastric mucin: Protective effect against oxygen radicals. Am J Physiol 1990;259:G681–686.PubMedGoogle Scholar
  30. 30.
    Carey MC, Small DM. The physical chemistry of cholesterol solubility in bile. Relationship to gallstone formation and dissolution in man. J Clin Invest 1978;61:998–1026.PubMedGoogle Scholar
  31. 31.
    Malet PF, Takabayashi A, Trotman BW, et al. Black and brown pigment gallstones differ in microstructure and microcomposition. Hepatology 1984;4:227–234.PubMedGoogle Scholar
  32. 32.
    Cahalane MJ, Neubrand MW, Carey MC. Physical-chemical pathogenesis of pigment gallstones. Semin Liver Dis 1988;8:317–328.PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • Hidetaka Higashijima
    • 1
  • Hitoshi Ichimiya
    • 1
  • Toru Nakano
    • 1
  • Hiroyuki Yamashita
    • 1
  • Shoji Kuroki
    • 1
  • Hiroshi Satoh
    • 1
  • Kazuo Chijiiwa
    • 1
  • Masao Tanaka
    • 1
  1. 1.First Department of SurgeryKyushu University Faculty of MedicineFukuokaJapan

Personalised recommendations