Abstract
Bile acid malabsorption has been shown to be associated with diarrhea in cases such as ileal resection Crohn's disease of the ileum, and radiation enteritis. The mechanisms of bile acid-induced diarrhea are not fully understood. Although the induction of colonic chloride secretion in response to bile acids has been extensively investigated, to date the direct effect of bile acids on intestinal chloride absorption has not been well defined. Therefore, the current studies were undertaken to investigate the effect of bile acids on the apical Cl(-)/OH(-) exchange process utilizing Caco2 monolayers as an in vitro cellular model. Cl(-)/OH(-) exchange activity was measured as DIDS-sensitive pH gradient-driven (36)Cl uptake. The results are summarized as follows: (i) short-term exposure (20 min) of Caco2 cells to taurodeoxycholate (TDC; 200 microM) and glycochenodeoxycholate (GCDC; 200 microM) acids significantly inhibited apical Cl(-)/OH(-) exchange (by approximately 60-70%); (ii) the Ca(2+) chelator BAPTA-AM blocked the inhibition by TDC; (iii) the reduction in Cl(-)/OH(-) exchange by TDC was reversed by the PKC inhibitor, chelerythrine chloride; (iv) functional and inhibitor studies indicated that TDC induced inhibition of Cl(-)/OH(-) exchange was mediated via the activation of the PKC beta I isoform; (v) the effect of TDC on apical Cl(-)/OH(-) exchange was completely blocked by the PI3 kinase inhibitor LY294002 (5 microM); and (vi) the PKA inhibitor, RpcAMP, had no effect on TDC induced inhibition of Cl(-)/OH(-) exchange. In conclusion, our studies provide direct evidence for inhibition of human intestinal apical Cl(-)/OH(-) exchange activity by bile acids via Ca(2+)-, PI3 kinase-, and PKC beta I-dependent pathways in Caco2 cells.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / pharmacology
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Alkaloids / pharmacology
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Antiporters / antagonists & inhibitors
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Antiporters / metabolism*
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Benzophenanthridines / pharmacology
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Caco-2 Cells
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Calcium / metabolism
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Chelating Agents / pharmacology
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Chlorides / metabolism
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Chromones / pharmacology
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Diarrhea / metabolism
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Diarrhea / physiopathology
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Dose-Response Relationship, Drug
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Egtazic Acid / analogs & derivatives
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Egtazic Acid / pharmacology
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Glycochenodeoxycholic Acid / metabolism
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Humans
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Hydrogen-Ion Concentration
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Intestinal Mucosa / drug effects
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Intestinal Mucosa / metabolism*
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Intestinal Mucosa / pathology
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Morpholines / pharmacology
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Organic Anion Transporters, Sodium-Dependent / metabolism
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Phosphatidylinositol 3-Kinases / metabolism
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Phosphoinositide-3 Kinase Inhibitors
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Protein Kinase C / antagonists & inhibitors
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Protein Kinase C / metabolism
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Protein Kinase C beta
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Protein Kinase Inhibitors / pharmacology
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Signal Transduction* / drug effects
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Sodium / metabolism
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Symporters / metabolism
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Taurodeoxycholic Acid / metabolism*
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Taurodeoxycholic Acid / pharmacology
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Time Factors
Substances
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Alkaloids
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Antiporters
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Benzophenanthridines
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Chelating Agents
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Chlorides
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Chromones
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Morpholines
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Organic Anion Transporters, Sodium-Dependent
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Phosphoinositide-3 Kinase Inhibitors
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Protein Kinase Inhibitors
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Symporters
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chloride-base exchanger
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1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester
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sodium-bile acid cotransporter
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2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
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Taurodeoxycholic Acid
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Egtazic Acid
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Glycochenodeoxycholic Acid
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Sodium
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chelerythrine
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Protein Kinase C
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Protein Kinase C beta
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4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid
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Calcium