The Role Of IL-4 And IL-13 Regulation MUC5B In Otitis Media With Effusion

Objective;This study will culture the HT29-MTX cells in inflammatory cytokine interleukin-4 (IL-4) or inflammatory cytokine interleukin-13 (IL-13). To investigate the role of the IL-4 and IL-13 in the regulation of mucin secretion. We will approach the pathogenesis of otitis media with effusion(OME).Methods:we cultured the HT-29 cells and then cultured with complete medium containing 10-5mol MTX. When the cell achieved> 80% of confluency, passage and assay it. we exposed the HT29-MTX cells to IL-4/IL-13 in a dose- dependent (5 ng/mL,10ng/mL,50ng/mL,100ng/mL,200 ng/mL) manner. After 24 hours,2 ml of media was aspirated and reserved for mucin quantification.RNA was isolated and reserved for RT-PCR. we exposed the HT 29-MTX cells to 50 ng/mL IL-4/IL-13 in a time- dependent (6 h,12 h,24 h,36h,48 h,72 h) manner.2 ml of media was aspirated and reserved for mucin quantification.RNA was isolated and reserved for RT-PCR. The controls was set in every group. The result was analyzed with Statistical.Result:(1) when the concentration of IL-4/IL-13 reached 10 ng/mL, the HT 29-MTX cells expressed the MUC5BmRNA and the protein of MUC5B increase comparison with the control (p<0.01)(2)After exposing the HT 29-MTX cells to 50ng/ml IL-4 12 h, as the time extending, which expressed the MUC5BmRNA and the protein of MUC5B increase comparison with the control (p<0.01).(3) After exposing the HT29-MTX cells to 50 ng/mL IL-13 24 h, as the time extending, which expressed the MUC5BmRNA and the protein of MUC5B increase comparison with the control (p<0.01)(4) No cytotoxic effects were observed at any concentration of IL-4/IL-13.Conclusion:(1) MUC5B take part in the pathological process of secretory otitis media.(2)In vitro the IL-4 and IL-13 up-regulation the secretion of MUC5B of goblet cells, which suggested IL-4 and IL-13 contribute to the Pathophysiology of OME. It may be a theoretical basis for clinical treatment OME.