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Tight junction integrity diminished by hypoxia, indomethacin, and hydrocortisone in Caco2 cell line (650.5)
Author(s) -
Barnes Rachel,
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.650.5
Subject(s) - barrier function , tight junction , hypoxia (environmental) , cytotoxicity , intestinal epithelium , viability assay , inflammation , apoptosis , epithelium , inflammatory bowel disease , intestinal permeability , chemistry , microbiology and biotechnology , pharmacology , immunology , biology , medicine , pathology , biochemistry , in vitro , disease , organic chemistry , oxygen
Background: Inflammation, hypoxia and loss of barrier function hallmark inflammatory bowel disease. Tight junctions play a vital role in the regulation of intestinal epithelial permeability. Indomethacin and hydrocortisone are used clinically as first line agents for inflammatory bowel disease. Objective: To examine the effects of indomethacin and hydrocortisone, concurrent with hypoxia, pro‐inflammatory cytokines, on tight junctional integrity and cell viability. 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). Epithelial barrier integrity assessed using solute flux assays, tight junction expression and localization analysis. Cell survivability evaluated by LDH quantification and cytotoxicity assays. Results: Epithelial barrier integrity was impaired under exposure of hypoxia alone; these effects compounded with exposure to pro‐inflammatory agents leading to increased cytotoxicity. Addition of anti‐inflammatory agents demonstrated improved cell survivability without improvement in barrier function as demonstrated by solute flux, confocal imaging, and cytotoxicity assays. Conclusions: Anti‐inflammatory agents may alter the regulation of intestinal epithelium cytotoxicity without improvement in barrier function. 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