title: Differential effects of deoxycholic acid and taurodeoxycholic acid on NF-kappa B signal transduction and IL-8 gene expression in colonic epithelial cells.
abstract: Several effects of bile acids (BAs) on colonic epithelial cells (CECs) have been described, including induction of proliferation and apoptosis. Some of these effects are mediated through activation of the NF-kappa B transcriptional system. In this study, we investigated the molecular mechanisms underlying the BA-induced gene expression in CECs. The human CEC line HT-29 and primary human CECs were treated with dilutions of salts of deoxycholic acid (DCA) and taurodeoxycholic acid (TDCA). NF-kappa B binding activity was analyzed with EMSA, RelA translocation with immunofluorescence, and I kappa B alpha- and RelA-phosphorylation with Western blot analysis. IL-8 mRNA and protein expression were assessed by quantitative PCR and ELISA. Functional impact of NF-kappa B activation was determined by blocking the proteasome activity with MG132 or by preventing IKK activity with a dominant-negative IKK beta delivered by adenoviral dominant-negative (dn) IKK beta (Ad5dnIKK beta). DCA and TDCA induced IL-8 expression in a dose- and time-dependent manner. It is interesting that DCA but not TDCA induced I kappa B alpha-phosphorylation, RelA translocation, and NF-kappa B binding activity. Accordingly, the proteasome inhibitor MG132 blocked DCA- but not TDCA-induced IL-8 gene expression. In contrast, TDCA-induced IL-8 gene expression correlated with enhanced RelA phosphorylation, which was blocked by Ad5dnIKK beta. Our data suggest that DCA-induced signal transduction mainly utilized the I kappa B degradation and RelA nuclear translocation pathway, whereas TDCA primarily induced IL-8 gene expression through RelA phosphorylation. These differences may have implications for the understanding of the pathophysiology of inflammation and carcinogenesis in the gut.
authors.affiliation: Department of Internal Medicine I, University of Regensburg, D-93042 Regensburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Mühlbauer
authors.forename: B
authors.initials: B
authors.lastname: Allard
authors.forename: A K
authors.initials: AK
authors.lastname: Bosserhoff
authors.forename: S
authors.initials: S
authors.lastname: Kiessling
authors.forename: H
authors.initials: H
authors.lastname: Herfarth
authors.forename: G
authors.initials: G
authors.lastname: Rogler
authors.forename: J
authors.initials: J
authors.lastname: Schölmerich
authors.forename: C
authors.initials: C
authors.lastname: Jobin
authors.forename: C
authors.initials: C
authors.lastname: Hellerbrand
mesh_terms: D001647:Bile Acids and Salts / Q000494:pharmacology; D002478:Cells, Cultured; D003106:Colon / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D003840:Deoxycholic Acid / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects*; D013657:Taurodeoxycholic Acid / Q000494:pharmacology*; D013997:Time Factors
chemical_list: D001647:Bile Acids and Salts; D016209:Interleukin-8; D016328:NF-kappa B; D012333:RNA, Messenger; D003840:Deoxycholic Acid; D013657:Taurodeoxycholic Acid