摘要
目的观察白细胞介素4(Interleukin-4,IL-4)和白细胞介素13(Interleukin-13,IL-13)刺激HT-29MTX细胞后,黏蛋白MUC5BmRNA和蛋白表达的变化,探讨分泌性中耳炎的发病机制。方法分别采用不同浓度梯度的IL-4、IL-13刺激HT-29MTX细胞,培养不同时间后,收集细胞培养基,提取细胞总RNA和蛋白,进行实时荧光定量PCR及Westernblot分析。结果 IL-4和IL-13浓度达到10ng/ml时,随着IL-4、IL-13浓度的增加,HT29-MTX细胞表达MUC5BmRNA增加且MUC5B蛋白分泌量增加,与空白组相比,有显著性差异(F=172.82、14.32,P<0.01)。HT-29MTX细胞暴露于含10ng/mlIL-4、IL-13的培养基时,分别在12h和24h后,随着时间的延长,HT-29MTX细胞表达MUC5BmRNA增加,且MUC5B蛋白分泌量增加,与空白组相比,差异有统计学意义(F=201.01、579.78,P<0.01)。结论 IL-4、IL-13能上调体外培养的杯状细胞MUC5B的分泌,提示其可能参与分泌性中耳炎的病理生理过程。
OBJECTIVE To explore the pathogenesis of Otitis Media With Effusion （OME） through observing regulation of IL-4 and IL-13 on MUC5B mRNA and secretion of protein by stimulation of HT-29MTX cell line. METHODS HT-29MTX cells were cultured with IL-4 or IL-13 in different concentration respectively for different time. Then culture media was collected for mucin analysis and total RNA and protein were extracted for quantity analysis by Real- time PCR and Western blot. RESULTS 1. When the concentration of IL-4 or IL-13 was over 10 ng/ml, MUC5B mRNA and MUC5B protein expressed by HT-29MTX cells increased with the concentration increase of IL-4 and IL- 13. It is statistically significant compared with control group （F=172.82, 14.32, P〈0.01） . 2. After exposing HT- 29MTX cells to 10ng/ml of IL-4 and IL13 for different durations, MUC5B mRNA and MUC5B protein increased after 12h and 24h respectively, with time extending. It is statistically significant compared with control group （F=201.01, 579.78, P〈0.01） . CONCLUSION IL-4 and IL-13 may be involved in the pathogenesis of OME by upregulation of MUC5B secretion in vitro.
出处
《中国耳鼻咽喉头颈外科》
2012年第12期645-648,共4页
Chinese Archives of Otolaryngology-Head and Neck Surgery
