title: Mechanism of bile salt-induced mucin secretion by cultured dog gallbladder epithelial cells.
abstract: 1. Hypersecretion of gallbladder mucin has been proposed to be a pathogenic factor in cholesterol gallstone formation. Using cultured gallbladder epithelial cells, we demonstrated that bile salts regulate mucin secretion by the gallbladder epithelium. In the present study we have investigated whether established second messenger pathways are involved in bile salt-induced mucin secretion. 2. The effect of activators and inhibitors on mucin secretion was studied by measuring the secretion of [3H]N-acetyl-D-glucosamine-labelled glycoproteins. Intracellular cAMP content of the cells was measured using a radioimmunoassay. 3. Incubation of the cells with 10 mM taurocholate did not increase the intracellular cAMP content (25.7 versus control 22.8 pmol of cAMP/mg of protein). No stimulation of mucin secretion was observed after incubation with 1-100 microM concentrations of the calcium ionophores ionomycin and A23187. The stimulatory effect of 10 mM tauroursodeoxycholate (TUDC) on mucin secretion could not be inhibited by the addition of EDTA. Activation of protein kinase C (PKC) by 1 microgram/ml phorbol 12-myristate 13-acetate (PMA) caused an increase in mucin secretion (342% versus control 100%), comparable with the effect of 40 mM TUDC. The effect of 10 ng/ml PMA could partially be inhibited by a concentration of 2 microM of the PKC inhibitor staurosporin. Staurosporin had no inhibitory effect on mucin secretion induced by TUDC. 4. In gallbladder epithelial cells bile salts do not stimulate mucin secretion via one of the classical signal transduction pathways. We hypothesize that bile salts act on mucin secretion via a direct interaction with the apical membrane.
authors.affiliation: Department of Gastroenterology, University of Amsterdam, Academic Medical Center, The Netherlands.
authors.forename: J H
authors.initials: JH
authors.lastname: Klinkspoor
authors.forename: G N
authors.initials: GN
authors.lastname: Tytgat
authors.forename: S P
authors.initials: SP
authors.lastname: Lee
authors.forename: A K
authors.initials: AK
authors.lastname: Groen
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D000818:Animals; D001647:Bile Acids and Salts / Q000494:pharmacology*; D000001:Calcimycin / Q000494:pharmacology; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D015232:Dinoprostone / Q000494:pharmacology; D004285:Dogs; D004492:Edetic Acid / Q000494:pharmacology; D004789:Enzyme Activation; D004848:Epithelium / Q000187:drug effects / Q000502:physiology; D005704:Gallbladder / Q000187:drug effects / Q000502:physiology*; D015759:Ionomycin / Q000494:pharmacology; D007700:Kinetics; D009077:Mucins / Q000096:biosynthesis / Q000378:metabolism*; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism; D015290:Second Messenger Systems; D013655:Taurochenodeoxycholic Acid / Q000494:pharmacology; D013656:Taurocholic Acid / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014316:Tritium
chemical_list: D001647:Bile Acids and Salts; D009077:Mucins; D014316:Tritium; D015240:Phorbol 12,13-Dibutyrate; D000001:Calcimycin; D013655:Taurochenodeoxycholic Acid; D015759:Ionomycin; D013656:Taurocholic Acid; C031655:ursodoxicoltaurine; D004492:Edetic Acid; D000242:Cyclic AMP; D011493:Protein Kinase C; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate; D000117:Acetylglucosamine