BACKGROUND

Inflammatory bowel disease (IBD) is characterized by a damaged intestinal epithelium barrier. Interferon regulatory factor 1 (IRF1) and osteopontin (OPN) regulate cell survival and growth in a variety of circumstances but their effects on the intestinal epithelium have not been elucidated. In this study, we sought to determine the effects of OPN on intestinal epithelial cells under conditions of tumor necrosis factor (TNF)-α-induced inflammation and whether IRF1 regulates OPN expression, the activation of downstream pathways, and inflammatory responses.

METHODS

The expression levels of OPN and IRF1 were assessed by immunohistochemical analyses of human IBD and experimental mouse colitis. The effects of IRF1 and OPN on inflammatory responses were investigated in vitro in NCM460 and Caco-2 cells stimulated by TNF-α. Changes in p-AKT, p-P38, and p-ERK levels were quantified by western blotting assays. The regulation of OPN expression by IRF1 was determined by luciferase activity and chromatin immunoprecipitation assays.

RESULTS

IRF1 was upregulated in human IBD and in the colon epithelium of mice with dextran sulfate sodium-induced colitis. Additionally, IRF1 was correlated with high-sensitivity C-reactive protein, erythrocyte sedimentation rate, Crohn's disease activity index, Crohn's disease endoscopic index of severity, and simple endoscopic score for Crohn's disease in Crohn's disease and with high-sensitivity C-reactive protein, erythrocyte sedimentation rate, Mayo score, Baron score, modified Baron score, Rachmilewitz score, ulcerative colitis endoscopic index of severity, ulcerative colitis colonoscopic index of severity, and disease duration in ulcerative colitis. The expression of OPN was significantly decreased in patients with IBD compared with controls and in dextran sulfate sodium-induced experimental colitis and was also inversely correlated with clinical and endoscopic activities in both Crohn's disease and ulcerative colitis. TNF-α treatment upregulated IRF1 and diminished OPN in both NCM460 and Caco-2 cells. The overexpression of OPN and rhOPN ameliorated the apoptosis induced by TNF-α, whereas the overexpression of IRF1 aggravated apoptosis, indicating opposite effects of OPN and IRF1 in inflamed epithelial cells. The luciferase and chromatin immunoprecipitation assays showed that IRF1 transcriptionally modulated the expression of OPN. TNF-α inhibited the OPN-induced upregulation of p-ERK, p-P38, and p-AKT.

CONCLUSIONS

Our data suggest that during intestinal inflammation, the TNF-α-mediated activation of IRF1 is related to the subsequent suppression of OPN expression, further reducing p-AKT, p-P38, and p-ERK activities and resulting in aggravation of the injury to intestinal epithelial cells.