Abstract
1. Bile salts stimulate mucin secretion by the gallbladder epithelium. We have investigated whether this stimulatory effect is due to a detergent effect of bile salts. 2. The bile salts taurocholic acid (TC) and tauroursodeoxycholic acid (TUDC) and the detergents Triton X-100 (12.5-400 microM) and Tween-20 (0.1-3.2 mM) were applied to monolayers of cultured dog gallbladder epithelial cells. Mucin secretion was studied by measuring the secretion of [3H]N-acetyl-d-glucosamine-labelled glycoproteins. We also attempted to alter the fluidity of the apical membrane of the cells through extraction of cholesterol with beta-cyclodextrin (2.5-15 mM). The effect on TUDC-induced mucin secretion was studied. Cell viability was assessed by measuring lactate dehydrogenase (LDH) leakage or 51Cr release. 3. In contrast with the bile salts, the detergents were not able to cause an increase in mucin secretion without causing concomitant cell lysis. Concentrations of detergent that increased mucin release (>100 microM Triton X-100, >0.8 mM Tween-20), caused increased LDH release. Incubation with beta-cyclodextrin resulted in effective extraction of cholesterol without causing an increase in 51Cr release. However, no effect of the presumed altered membrane fluidity on TUDC (10 mM)-induced mucin secretion was observed. 4. The stimulatory effect of bile salts on mucin secretion by gallbladder epithelial cells is not affected by the fluidity of the apical membrane of the cells and also cannot be mimicked by other detergents. We conclude that the ability of bile salts to cause mucin secretion by the gallbladder epithelium is not determined by their detergent properties.
Full Text
The Full Text of this article is available as a PDF (230.3 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- AMADOR E., DORFMAN L. E., WACKER W. E. SERUM LACTIC DEHYDROGENASE ACTIVITY: AN ANALYTICAL ASSESSMENT OF CURRENT ASSAYS. Clin Chem. 1963 Aug;12:391–399. [PubMed] [Google Scholar]
- Beuers U., Nathanson M. H., Boyer J. L. Effects of tauroursodeoxycholic acid on cytosolic Ca2+ signals in isolated rat hepatocytes. Gastroenterology. 1993 Feb;104(2):604–612. doi: 10.1016/0016-5085(93)90433-d. [DOI] [PubMed] [Google Scholar]
- Beuers U., Nathanson M. H., Isales C. M., Boyer J. L. Tauroursodeoxycholic acid stimulates hepatocellular exocytosis and mobilizes extracellular Ca++ mechanisms defective in cholestasis. J Clin Invest. 1993 Dec;92(6):2984–2993. doi: 10.1172/JCI116921. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Beuers U., Throckmorton D. C., Anderson M. S., Isales C. M., Thasler W., Kullak-Ublick G. A., Sauter G., Koebe H. G., Paumgartner G., Boyer J. L. Tauroursodeoxycholic acid activates protein kinase C in isolated rat hepatocytes. Gastroenterology. 1996 May;110(5):1553–1563. doi: 10.1053/gast.1996.v110.pm8613063. [DOI] [PubMed] [Google Scholar]
- Bouscarel B., Ceryak S., Gettys T. W., Fromm H., Noonan F. Alteration of cAMP-mediated hormonal responsiveness by bile acids in cells of nonhepatic origin. Am J Physiol. 1995 Jun;268(6 Pt 1):G908–G916. doi: 10.1152/ajpgi.1995.268.6.G908. [DOI] [PubMed] [Google Scholar]
- Bouscarel B., Gettys T. W., Fromm H., Dubner H. Ursodeoxycholic acid inhibits glucagon-induced cAMP formation in hamster hepatocytes: a role for PKC. Am J Physiol. 1995 Feb;268(2 Pt 1):G300–G310. doi: 10.1152/ajpgi.1995.268.2.G300. [DOI] [PubMed] [Google Scholar]
- Coleman R. Bile salts and biliary lipids. Biochem Soc Trans. 1987 Dec;15 (Suppl):68S–80S. [PubMed] [Google Scholar]
- Coleman R., Lowe P. J., Billington D. Membrane lipid composition and susceptibility to bile salt damage. Biochim Biophys Acta. 1980 Jun 20;599(1):294–300. doi: 10.1016/0005-2736(80)90075-9. [DOI] [PubMed] [Google Scholar]
- Combettes L., Berthon B., Doucet E., Erlinger S., Claret M. Bile acids mobilise internal Ca2+ independently of external Ca2+ in rat hepatocytes. Eur J Biochem. 1990 Jul 5;190(3):619–623. doi: 10.1111/j.1432-1033.1990.tb15617.x. [DOI] [PubMed] [Google Scholar]
- Coquil J. F., Berthon B., Chomiki N., Combettes L., Jourdon P., Schteingart C., Erlinger S., Claret M. Effects of taurolithocholate, a Ca2(+)-mobilizing agent, on cell Ca2(+) in rat hepatocytes, human platelets and neuroblastoma NG108-15 cell line. Biochem J. 1991 Jan 1;273(Pt 1):153–160. doi: 10.1042/bj2730153. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Devor D. C., Sekar M. C., Frizzell R. A., Duffey M. E. Taurodeoxycholate activates potassium and chloride conductances via an IP3-mediated release of calcium from intracellular stores in a colonic cell line (T84) J Clin Invest. 1993 Nov;92(5):2173–2181. doi: 10.1172/JCI116819. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dray-Charier N., Paul A., Combettes L., Bouin M., Mergey M., Balladur P., Capeau J., Housset C. Regulation of mucin secretion in human gallbladder epithelial cells: predominant role of calcium and protein kinase C. Gastroenterology. 1997 Mar;112(3):978–990. doi: 10.1053/gast.1997.v112.pm9041261. [DOI] [PubMed] [Google Scholar]
- Hayashi A., Lee S. P. Bidirectional transport of cholesterol between gallbladder epithelial cells and model bile. Am J Physiol. 1996 Sep;271(3 Pt 1):G410–G414. doi: 10.1152/ajpgi.1996.271.3.G410. [DOI] [PubMed] [Google Scholar]
- Huang X. P., Fan X. T., Desjeux J. F., Castagna M. Bile acids, non-phorbol-ester-type tumor promoters, stimulate the phosphorylation of protein kinase C substrates in human platelets and colon cell line HT29. Int J Cancer. 1992 Sep 30;52(3):444–450. doi: 10.1002/ijc.2910520319. [DOI] [PubMed] [Google Scholar]
- Kilsdonk E. P., Yancey P. G., Stoudt G. W., Bangerter F. W., Johnson W. J., Phillips M. C., Rothblat G. H. Cellular cholesterol efflux mediated by cyclodextrins. J Biol Chem. 1995 Jul 21;270(29):17250–17256. doi: 10.1074/jbc.270.29.17250. [DOI] [PubMed] [Google Scholar]
- Klein U., Gimpl G., Fahrenholz F. Alteration of the myometrial plasma membrane cholesterol content with beta-cyclodextrin modulates the binding affinity of the oxytocin receptor. Biochemistry. 1995 Oct 24;34(42):13784–13793. doi: 10.1021/bi00042a009. [DOI] [PubMed] [Google Scholar]
- Klinkspoor J. H., Kuver R., Savard C. E., Oda D., Azzouz H., Tytgat G. N., Groen A. K., Lee S. P. Model bile and bile salts accelerate mucin secretion by cultured dog gallbladder epithelial cells. Gastroenterology. 1995 Jul;109(1):264–274. doi: 10.1016/0016-5085(95)90293-7. [DOI] [PubMed] [Google Scholar]
- Klinkspoor J. H., Tytgat G. N., Lee S. P., Groen A. K. Mechanism of bile salt-induced mucin secretion by cultured dog gallbladder epithelial cells. Biochem J. 1996 Jun 15;316(Pt 3):873–877. doi: 10.1042/bj3160873. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kuver R., Savard C., Oda D., Lee S. P. PGE generates intracellular cAMP and accelerates mucin secretion by cultured dog gallbladder epithelial cells. Am J Physiol. 1994 Dec;267(6 Pt 1):G998–1003. doi: 10.1152/ajpgi.1994.267.6.G998. [DOI] [PubMed] [Google Scholar]
- Lowe P. J., Coleman R. Membrane fluidity and bile salt damage. Biochim Biophys Acta. 1981 Jan 8;640(1):55–65. doi: 10.1016/0005-2736(81)90531-9. [DOI] [PubMed] [Google Scholar]
- Oda D., Lee S. P., Hayashi A. Long-term culture and partial characterization of dog gallbladder epithelial cells. Lab Invest. 1991 May;64(5):682–692. [PubMed] [Google Scholar]
- Ohtani Y., Irie T., Uekama K., Fukunaga K., Pitha J. Differential effects of alpha-, beta- and gamma-cyclodextrins on human erythrocytes. Eur J Biochem. 1989 Dec 8;186(1-2):17–22. doi: 10.1111/j.1432-1033.1989.tb15171.x. [DOI] [PubMed] [Google Scholar]
- Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
- Stravitz R. T., Vlahcevic Z. R., Gurley E. C., Hylemon P. B. Repression of cholesterol 7 alpha-hydroxylase transcription by bile acids is mediated through protein kinase C in primary cultures of rat hepatocytes. J Lipid Res. 1995 Jun;36(6):1359–1369. [PubMed] [Google Scholar]
- Velardi A. L., Groen A. K., Elferink R. P., van der Meer R., Palasciano G., Tytgat G. N. Cell type-dependent effect of phospholipid and cholesterol on bile salt cytotoxicity. Gastroenterology. 1991 Aug;101(2):457–464. doi: 10.1016/0016-5085(91)90025-g. [DOI] [PubMed] [Google Scholar]
- Zimniak P., Little J. M., Radominska A., Oelberg D. G., Anwer M. S., Lester R. Taurine-conjugated bile acids act as Ca2+ ionophores. Biochemistry. 1991 Sep 3;30(35):8598–8604. doi: 10.1021/bi00099a015. [DOI] [PubMed] [Google Scholar]
