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Intestinal barrier integrity diminished by hypoxia, indomethacin, and hydrocortisone in Caco2 cell line (650.12)
Author(s) -
Corley Katherine,
Carr Nick,
King Jonathan
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.650.12
Subject(s) - barrier function , tight junction , hypoxia (environmental) , intestinal epithelium , inflammation , intestinal permeability , chemistry , microbiology and biotechnology , epithelium , medicine , biology , pathology , organic chemistry , oxygen
Background: Concomitant use of hydrocortisone and the nonspecific cyclooxygenase (COX)‐inhibitor indomethacin increases the risk for spontaneous intestinal perforation in extremely low birth weight preterm infants. The regulation of the eicosanoid pathway and inflammation play a large role in maintaining intestinal barrier integrity. Objective: Test the hypothesis that the dysregulation of the PGE2/EP pathway leads to altered intestinal barrier integrity, impacting cell survival and tight junction function. Methods: A Caco‐2 cell line was utilized in the setting of normoxia (160 torr) and hypoxia (20 torr) under varying conditions of pro‐inflammatory agents TNF (10ng/mL), IFN‐gamma (10ng/mL)) and anti‐inflammatory agents of indomethacin (50uM) and hydrocortisone (50ng/mL). Barrier function was assessed using transepithelial electrical resistance (TER) measurements and solute flux assays. Cell survivability and morphology was evaluated with direct confocal microscopy as well as indirect LDH quantification, cytotoxicity assays. Results: The concomitant exposure of the Caco‐2 cell monolayer to hypoxia, pro‐inflammatory and anti‐inflammatory agents demonstrated abnormalities of barrier integrity, cell morphology, and impairment of tight junction formation on confocal microscopy, not seen in normoxia conditions Conclusions: Anti‐inflammatory agents may alter the regulation of intestinal epithelium cytotoxicity without improvement in tight junction integrity. These findings indicate the possible role of indomethacin, hydrocortisone in altering cell survival mechanisms leading to cellular necrosis vs. apoptosis under hypoxic, pro‐inflammatory environments. Grant Funding Source : Supported in part by NSF‐MRI #1229702