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    Bacterial lipopolysaccharide promotes profibrotic activation of intestinal fibroblasts.

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    Authors
    Burke, J P
    Cunningham, M F
    Watson, R W G
    Docherty, N G
    Coffey, J C
    O'Connell, P R
    Affiliation
    Department of Surgery, St Vincent's University Hospital, Dublin, Ireland.
    Issue Date
    2012-02-01T10:28:56Z
    MeSH
    Aged
    Aged, 80 and over
    Cells, Cultured
    Colonic Neoplasms/metabolism
    Connective Tissue Growth Factor/biosynthesis
    Fibroblasts/*drug effects/metabolism
    Humans
    I-kappa B Kinase/metabolism
    Lipopolysaccharides/*pharmacology
    Middle Aged
    NF-kappa B/metabolism
    Smad7 Protein/metabolism
    Toll-Like Receptor 4/*metabolism
    Transforming Growth Factor beta1/*pharmacology
    Wound Healing/*physiology
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    Citation
    Br J Surg. 2010 Jul;97(7):1126-34.
    Journal
    The British journal of surgery
    URI
    http://hdl.handle.net/10147/207477
    DOI
    10.1002/bjs.7045
    PubMed ID
    20632282
    Abstract
    BACKGROUND: Fibroblasts play a critical role in intestinal wound healing. Lipopolysaccharide (LPS) is a cell wall component of commensal gut bacteria. The effects of LPS on intestinal fibroblast activation were characterized. METHODS: Expression of the LPS receptor, toll-like receptor (TLR) 4, was assessed in cultured primary human intestinal fibroblasts using flow cytometry and confocal microscopy. Fibroblasts were treated with LPS and/or transforming growth factor (TGF) beta1. Nuclear factor kappaB (NFkappaB) pathway activation was assessed by inhibitory kappaBalpha (IkappaBalpha) degradation and NFkappaB promoter activity. Fibroblast contractility was measured using a fibroblast-populated collagen lattice. Smad-7, a negative regulator of TGF-beta1 signalling, and connective tissue growth factor (CTGF) expression were assessed using reverse transcriptase-polymerase chain reaction and western blot. The NFkappaB pathway was inhibited by IkappaBalpha transfection. RESULTS: TLR-4 was present on the surface of intestinal fibroblasts. LPS treatment of fibroblasts induced IkappaBalpha degradation, enhanced NFkappaB promoter activity and increased collagen contraction. Pretreatment with LPS (before TGF-beta1) significantly increased CTGF production relative to treatment with TGF-beta1 alone. LPS reduced whereas TGF-beta1 increased smad-7 expression. Transfection with an IkappaBalpha plasmid enhanced basal smad-7 expression. CONCLUSION: Intestinal fibroblasts express TLR-4 and respond to LPS by activating NFkappaB and inducing collagen contraction. LPS acts in concert with TGF-beta1 to induce CTGF. LPS reduces the expression of the TGF-beta1 inhibitor, smad-7.
    Language
    eng
    ISSN
    1365-2168 (Electronic)
    0007-1323 (Linking)
    ae974a485f413a2113503eed53cd6c53
    10.1002/bjs.7045
    Scopus Count
    Collections
    St. Vincent's University Hospital

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