IL-4和IL-13在分泌性中耳炎中对MUC5B的调节作用

发布时间：2018-06-18 01:12

本文选题：IL-4 + IL-13　；参考：《遵义医学院》2011年硕士论文

【摘要】：目的：本研究拟通过炎症细胞因子IL-4、IL-13刺激HT-29MTX(一种可分泌粘蛋白的杯状细胞)细胞,观察其对粘蛋白的调节作用,从而进一步探讨分泌性中耳炎的发病机制。方法：HT-29细胞株复苏培养后,用10-5 mol MTX处理,待细胞生长至80%融合时,传代并鉴定细胞。IL-4、IL-13分别以不同浓度梯度(5 ng/mL、10ng/mL、50 ng/mL、100 ng/mL、200 ng/mL)刺激HT 29-MTX细胞分泌,培养24 h后,收集细胞培养基,并提取细胞RNA,进行粘蛋白及其RNA定量分析。用50ng/mL的IL-4、IL-13不同时间长度(6 h、12 h、24 h、36 h、48 h、72 h)刺激HT29MTX细胞分泌,收集培养基,并提取细胞RNA,进行粘蛋白及其RNA定量分析。每个浓度梯度组及时间梯度组均设相应对照组(加入PBS组)。统计学分析结果。结果：(1)培养于含有IL-4、IL-13培养基的HT29-MTX细胞,当IL-4、IL-13浓度达到10 ng/mL时,随着IL-4、IL-13浓度的增加,HT 29-MTX细胞表达MUC5BmRNA增加且MUC5B蛋白分泌量增加,与空白组相比(p0.01),差异有统计学意义。(2)HT 29-MTX细胞暴露于50 ng/mL IL-4的培养基时,在12h后,随着时间的延长,HT 29-MTX细胞表达MUC5BmRNA增加且MUC5B蛋白分泌量增加,与空白组相比(p0.01),差异有统计学意义。(3)HT 29-MTX细胞暴露于50 ng/mL IL-13的培养基时,在24 h后,随着时间的延长,其表达增加且MUC5B蛋白分泌量增加,与空白组相比(p0.01),差异有统计学意义。(4)在细胞培养整个过程中,加入各种浓度的IL-4、IL-13无出现细胞死亡现象,无表现出细胞毒性。结论：(1)MUC5B参与分泌性中耳炎的病理过程。(2)IL-4、IL-13上调体外培养的杯状细胞MUC5B的分泌,提示其可能参与慢性分泌性中耳炎的病理生理过程,为慢性分泌性中耳炎的治疗提供理论依据。
[Abstract]:Aim: to explore the mechanism of secretory otitis media by stimulating HT-29 MTX cells (goblet cells) with inflammatory cytokines IL-4 and IL-13. Methods after resuscitation culture, the cell line was resuscitated and treated with 10-5 mol MTX. When the cell grew to 80% fusion, the cell line. IL-4 IL-13 was subcultured and identified. The HT29-MTX cells were stimulated with different concentration gradient of 5 ng / mL ~ (10) ng / m ~ (-1) ~ (50) ng / m ~ (-1) ~ 100 ng / m ~ (-1). After 24 h of culture, the cell culture medium was collected. Cell RNAs were extracted for quantitative analysis of mucin and its RNA. HT29MTX cells were stimulated with 50 ng / mL IL-4 IL-13 for 6 h ~ 12 h ~ 24 h ~ 36 h ~ 48 h ~ 72 h). The culture medium was collected and cell RNAs were extracted for quantitative analysis of mucin and its RNA. Each concentration gradient group and time gradient group were divided into corresponding control group (add PBS group). Statistical analysis results. Results when the concentration of IL-4 / IL-13 reached 10 ng / mL, the expression of MUC5B mRNA and the secretion of MUC5B protein in HT29-MTX cells increased with the increase of IL-4 / IL-13 concentration. Compared with the control group, the difference was statistically significant. When the cells were exposed to 50 ng / mL IL-4 medium, the expression of MUC5B mRNA and the secretion of MUC5B protein were increased with the prolongation of time. Compared with the control group, the difference was statistically significant. When the cells were exposed to 50 ng / mL IL-13 medium, the expression of MUC5B protein increased and the secretion of MUC5B protein increased with the prolongation of time after 24 h. Compared with the control group, the difference was statistically significant (P < 0.05). During the whole process of cell culture, no cell death and no cytotoxicity were found in the addition of IL-4 / IL-13 at various concentrations. Conclusion MUC5B is involved in the pathological process of secretory otitis media. IL-4 / IL-13 up-regulates the secretion of MUC5B in goblet cells in vitro, which suggests that MUC5B may participate in the pathophysiological process of chronic secretory otitis media and provide a theoretical basis for the treatment of chronic secretory otitis media.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R764.21

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