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Isolation of mucin from human hepatic bile and its induced effects on precipitation of cholesterol and calcium carbonatein vitro

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Abstract

biliary mucin was isolated from human hepatic bile, and its induced effects on the appearance time of cholesterol monohydrate crystals (nucleation time) and on the precipitation of calcium carbonate were studiedin vitro to examine the possible significance of mucin for ductular gallstone formation. Mucin was isolated by gel filtration on Sepharose CL-4B and a subsequent CsCl density gradient ultracentrifugation. Mucin thus obtained had a high purity as shown by a high-molecular-weight band on SDS-polyacrylamide gel electrophoresis and by the compatible amino acid composition with mucin purified from the gallbladder. The mucin at as low a concentration as 100 μg/ml significantly shortened the cholesterol nucleation time in the supersaturated model bile, mimicking human hepatic bile. On the other hand, the addition of mucin inhibited calcium carbonate precipitationin vitro. Taking account of that both cholesterol and calcium salts are major constituents of ductular gallstones, we conclude that biliary mucin is likely to play an important regulating role in the formation of ductular stones.

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References

  1. Verschure JCM: Electro-chromatograms of human bile. Clin Chim Acta 1:38–48, 1956

    Google Scholar 

  2. Bouchier IAD, Cooperband SR, El Kodsi BM: Mucous substances and viscosity of normal and pathological human bile. Gastroenterology 49:343–353, 1965

    Google Scholar 

  3. Womack NA, Zeppa R, Irvin GL III: The anatomy of gallstones. Ann Surg 157:670–686, 1963

    Google Scholar 

  4. Maki T, Matsushiro T, Suzuki N, Nakamura N: Role of sulfated glycoprotein in gallstone formation. Surg Gynecol Obstet 132:846–854, 1971

    Google Scholar 

  5. LaMont JT, Ventola AS, Trotman BW, Soloway RD: Mucin glycoprotein content of human pigment gallstones. Hepatology 3:377–382, 1983

    Google Scholar 

  6. Pearson JP, Foster SNE: Mucous glycoprotein content of human cholesterol gallstones. Digestion 36:132–140, 1987

    Google Scholar 

  7. Lee SP, LaMont JT, Carey MC: Role of gallbladder mucus hypersecretion in the evolution of cholesterol gallstones. J Clin Invest 67:1712–1723, 1981

    Google Scholar 

  8. Levy PF, Smith BF, LaMont JT: Human gallbladder mucin accelerates nucleation of cholesterol in artificial bile. Gastroenterology 87:270–275, 1987

    Google Scholar 

  9. Smith BF: Human gallbladder mucin binds biliary lipids and promotes cholesterol crystal nucleation in model bile. J Lipid Res 28:1088–1097, 1987

    Google Scholar 

  10. Nakayama F: Quantitative microanalysis of gallstones. J Lab Clin Med 72:602–611, 1968

    Google Scholar 

  11. Lee SP, Nicholls JF: Diffusion of charged ions in mucus gel: Effect of net charge. Biorheology 24:565–570, 1987

    Google Scholar 

  12. Forstner JF, Forstner GG: Calcium binding to intestinal goblet cell mucin. Biochim Biophys Acta 386:283–292, 1975

    Google Scholar 

  13. Schiffman ML, Smith BF, Hirsch JI, Moore EW: Ca++ binding to bovine gallbladder mucin: high affinity but low capacity. Hepatology 10:601, 1989 (abstract)

    Google Scholar 

  14. Nagashima H, Masubuchi M, Yosizawa Z: Coagulating effects on calcium carbonate of sulfated glycoproteins isolated from pathological human bile. J Biochem 75:779–786, 1974

    Google Scholar 

  15. Carey MC, Small DM: The physical chemistry of cholesterol solubility in bile. J Clin Invest 61:998–1026, 1978

    Google Scholar 

  16. Rege RV, Moore EW: Pathogenesis of calcium-containing gallstones. Canine ductular bile, but not gallbladder bile, is supersaturated with calcium carbonate. J Clin Invest 77:21–26, 1986

    Google Scholar 

  17. Pearson JP, Kaura R, Taylor W, Allen A: The composition and polymeric structure of mucus glycoprotein from human gallbladder bile. Biochim Biophys Acta 706:221–228, 1982

    Google Scholar 

  18. Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F: Colorimetric method of determination of sugars and relate substances. Anal Chem 28:350–356, 1956

    Google Scholar 

  19. Starkey BJ, Snary D, Allen A: Characterization of gastric mucoproteins isolated by equilibrium density-gradient centrifugation in caesium chloride. Biochem J 141:633–639, 1974

    Google Scholar 

  20. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685, 1970

    Google Scholar 

  21. Wray W, Boulikas T, Wray VP, Hancock R: Silver staining of proteins in polyacrylamide gels. Anal Biochem 118:197–203, 1981

    Google Scholar 

  22. Fairbanks G, Steck TL, Wallach DFH: Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10:2606–2616, 1971

    Google Scholar 

  23. Kibe A, Dudley MA, Halpern Z, Lynn MP, Breuer AC, Holzbach RT: Factors affecting cholesterol monohydrate crystal nucleation time in model systems of supersaturated bile. J Lipid Res 26:1102–1111, 1985

    Google Scholar 

  24. Carey MC: Critical tables for calculating the cholesterol saturation of native bile. J Lipid Res 19:945–955, 1978

    Google Scholar 

  25. Kibe A, Holzbach RT, LaRusso NF, Mao SJT: Inhibition of cholesterol crystal formation by apolipoproteins A-I and A-II in model systems of supersaturated bile: Implications for gallstone pathogenesis in man. Science 225:514–516, 1984

    Google Scholar 

  26. Fukudome K, Chijiiwa K, Furusawa T, Nakayama F: Effect of albumin on the solubility of cholesterol in bile. Biochim Biophys Acta 922:155–161, 1987

    Google Scholar 

  27. Holan KR, Holzbach RT, Hermann RE, Cooperman AM, Claffey WJ: Nucleation time: A key factor in the pathogenesis of cholesterol gallstone disease. Gastroenterology 77:611–617, 1979

    Google Scholar 

  28. Dawes LG, Ostrow JD, Shimizu S, Moore EW, Rege RV: Inhibition of CaCO3 precipitation by canine gallbladder bile. Gastroenterology 94:A534, 1988 (abstract)

    Google Scholar 

  29. Smith BF, LaMont JT: Hydrophobic binding properties of bovine gallbladder mucin. J Biol Chem 259:12170–12177, 1984

    Google Scholar 

  30. Terada T, Nakanuma Y: Morphological examination of intrahepatic bile ducts in hepatolithiasis. Virchows Arch (A) 413:167–176, 1988

    Google Scholar 

  31. Yamasaki T, Nakayama F, Tamura S, Endo M: Characterization of mucin in the hepatic bile of the patients with intrahepatic pigment stones. J Gastroenterol Hepatol 7:36–41, 1992

    Google Scholar 

  32. Shimizu S, Sabsay B, Veis A, Ostrow JD, Rege RV, Dawes LG: Isolation of an acidic protein from cholesterol gallstones, which inhibits the precipitation of calcium carbonatein vitro. J Clin Invest 84:1990–1996, 1989

    Google Scholar 

  33. Moore EW: The role of calcium in the pathogenesis of gallstones: Ca++ electrode studies of model bile salt solutions and other biologic systems. Hepatology 4:228S-243S, 1984

    Google Scholar 

  34. Qiu SM, Crowther RS, Wen G, Soloway RD: Calcium hydroxyapatite precipitation in vitro is influenced by human gallbladder mucin. Gastroenterology 100:A787, 1991 (abstract)

    Google Scholar 

  35. Yamashita N, Yanagisawa J, Nakayama F: Composition of intrahepatic calculi. Dig Dis Sci 33:449–453, 1988

    Google Scholar 

  36. Tabata M, Nakayama F: Bacteria and gallstones. Dig Dis Sci 26:218–224, 1981

    Google Scholar 

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Authors and Affiliations

  1. Department of Surgery I, Kyushu University Faculty of Medicine, 3-1-1 maedashi, Higashi-ku, 812, Fukuoka, Japan

    Toru Yamasaki MD,  Kazuo Chijiiwa MD & Masahiko Endo MD

  2. Department of Biochemistry, Hirosaki University School of Medicine, Hirosaki, Japan

    Toru Yamasaki MD,  Kazuo Chijiiwa MD & Masahiko Endo MD

Authors
  1. Toru Yamasaki MD
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  2. Kazuo Chijiiwa MD
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  3. Masahiko Endo MD
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Yamasaki, T., Chijiiwa, K. & Endo, M. Isolation of mucin from human hepatic bile and its induced effects on precipitation of cholesterol and calcium carbonatein vitro . Digest Dis Sci 38, 909–915 (1993). https://doi.org/10.1007/BF01295919

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  • DOI: https://doi.org/10.1007/BF01295919

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