Bile acids modulate tight junction structure and barrier function of Caco-2 monolayers via EGFR activation

• DOI: 10.1152/ajpgi.00043.2007
• Publication Date: 2008-02-01T01:59:03Z
• Authors: Francesco Raimondi; Pasquale Santoro; Maria Vittoria Ba...; Serena Pappacoda; Maria Luisa Barretta; Merlin Nanayakkara; Carmela Apicella; Letizia Capasso; Roberto Paludetto
• Keywords: 0301 basic medicine; EGFR; tight junction; Cholic Acid; Chenodeoxycholic Acid; Permeability; Bile Acids and Salts; 03 medical and health sciences; Organophosphorus Compounds; Occludin; Electric Impedance; Organometallic Compounds; Humans; Intestinal Mucosa; Phosphorylation; 0303 health sciences; Epidermal Growth Factor; Bile acids; EGFR; tight junction; Antibodies, Monoclonal; Membrane Proteins; Dextrans; Bile acids; 3. Good health; Enzyme Activation; ErbB Receptors; Kinetics; Caco-2 Cells; Deoxycholic Acid

Intestinal and systemic illnesses have been linked to increased gut permeability. Bile acids, whose luminal profile can be altered in human disease, modulate intestinal paracellular permeability. We investigated the mechanism by which selected bile acids increase gut permeability using a validated in vitro model. Human intestinal Caco-2 cells were grown in monolayers and challenged with a panel of bile acids. Transepithelial electrical resistance and luminal-to-basolateral fluxes of 10-kDa Cascade blue-conjugated dextran were used to monitor paracellular permeability. Immunoprecipitation and immunoblot analyses were employed to investigate the intracellular pathway. Redistribution of tight junction proteins was studied by confocal laser microscopy. Micromolar concentrations of cholic acid, deoxycholic acid (DCA), and chenodeoxycholic acid (CDCA) but not ursodeoxycholic acid decreased transepithelial electrical resistance and increased dextran flux in a reversible fashion. Coincubation of 50 μM CDCA or DCA with EGF, anti-EGF monoclonal antibody, or specific src inhibitor 4-Amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP-2) abolished the effect. A concentration of 50 μM of either CDCA or DCA also induced EGF receptor phosphorylation, occludin dephosphorylation, and occludin redistribution at the tight junction level in the same time frame and in a reversible fashion. We conclude that selected bile acids modulate intestinal permeability via EGF receptor autophosphorylation, occludin dephosphorylation, and rearrangement at the tight junction level. The effect is mediated by the src family kinases and is abolished by EGF treatment. These data also support the role of bile acids in the genesis of necrotizing enterocolitis and the protective effect of EGF treatment.