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Objective: This study aimed to inflammatory cytokines IL-4, IL-13 stimulation of HT-29MTX ( one kind of mucin secreting goblet cells ) cells and observe its effect on the regulation of mucin , which further explore the secretory otitis media pathogenesis . Method : HT-29 cells were cultured recovery after treatment with 10-5 mol MTX , cells were grown to 80% until confluence, and passaged cells were identified . IL-4, IL-13 , respectively, in different concentrations (5 ng / mL, 10ng/mL, 50 ng / mL, 100 ng / mL, 200 ng / mL) stimulated HT 29-MTX cells , cultured for 24 h, collecting the cell culture medium and cell extract RNA, and RNA for quantitative analysis of mucin . With 50ng/mL of IL-4, IL-13 of different length of time (6 h, 12 h, 24 h, 36 h, 48 h, 72 h) to stimulate HT29MTX cells , the medium was collected and extracted cellular RNA, for sticky quantitative analysis of protein and RNA . Each concentration gradient group and the time gradient group are located corresponding control groups ( adding PBS group ) . Statistical analysis. Results : ( 1 ) containing cultured in IL-4, IL-13 medium of HT29-MTX cells when IL-4, IL-13 concentration of 10 ng / mL , with the IL-4, IL-13 concentration increase , HT 29-MTX cells MUC5BmRNA increases and MUC5B protein secretion increased, compared with the control group (p lt; 0.01), the difference was statistically significant. (2) HT 29-MTX cells were exposed to 50 ng / mL IL-4 of the culture medium , at 12h , with the extension of time , HT 29-MTX cells increased and MUC5B MUC5BmRNA increased protein secretion , with the control group compared (p lt; 0.01), the difference was statistically significant. (3) HT 29-MTX cells were exposed to 50 ng / mL IL-13 in culture medium , after 24 h , with the time , and its expression is increased and MUC5B increased protein secretion , compared with the control group (p lt; 0.01), the difference was statistically significant. ( 4 ) throughout the process in the cell culture , adding various concentrations of IL-4, IL-13 is no evidence of cell death phenomenon, no exhibit cytotoxicity. Conclusions : (1) MUC5B involved in the pathological process of secretory otitis media . (2) IL-4, IL-13 increased goblet cells in vitro MUC5B secretion , suggesting that it may be involved in chronic otitis media with effusion in the pathophysiological process for the treatment of chronic otitis media with effusion provide a theoretical basis .
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