Proinflammatory effects and molecular mechanisms of interleukin-17 in intestinal epithelial cell line HT-29

AIM: To evaluate the proinflammatory effects and molecular mechanisms of interleukin (IL)-17 in intestinal epithelial cell line HT-29. METHODS: HT-29 cells were cultured with IL-17, tumor necrosis factor (TNF)-alpha, or the combination of both IL-17 and TNF-alpha. Real-time PCR and Western blot were used to measure the gene expression levels of neutro-phil chemokines CXCL1, CXCL2, CXCL5, CXCL6, IL-8 and TH-17 cell chemokine CCL20, the phosphorylation levels of p38 and TNF-alpha, and the expression level of IL-8, after using the p38 inhibitor in HT-29 cells. The stable Act1 knockdown HT-29 cell line was established to further test the phosphorylation changes of p38, after using IL-17 and TNF-alpha. RESULTS: After HT-29 cells were cultured with IL-17 and TNF-alpha, the expression levels of neutrophil chemokines (CXCL1, CXCL2, CXCL5, CXCL6, IL-8) and Th17 chemokine (CCL20) significantly improved (24.96 +/- 2.53, 28.47 +/- 2.87, 38.08 +/- 2.72, 33.47 +/- 2.41, 31.7 +/- 2.38, 44.37 +/- 2.73, respectively), and the differences were all statistically significant (P < 0.01). Western blot results showed that IL-17 obviously enhanced the phosphorylation level of p38, which was induced by TNF-alpha. Compared with the control group, the expression level of IL-8 significantly declined (9.47 +/- 1.36 vs 3.06 +/- 0.67, P < 0.01) when TH-29 cells were cultured with IL-17 and TNF-alpha. p38 inhibition assay showed that the p38 pathway played an essential role in the inflammatory response induced by IL-17. p38 phosphorylation levels could not be changed after using IL-17 and TNF-alpha in the stable Act1 knockdown HT-29 cell line. CONCLUSION: IL-17 significantly promoted the gene expression levels of TNF-alpha-induced neutrophil chemokines and Th17 cell chemokine. It is obvious that IL-17 and TNF-alpha have synergistic effects on p38. (C) 2014 Baishideng Publishing Group Inc. All rights reserved.