摘要
目的探讨miR-34c对慢性鼻-鼻窦炎伴鼻息肉(chronic rhinosinusitis with nasal polyps,CRSwNP)鼻上皮纤毛细胞分化的影响及其调控机制。方法获取正常对照及CRSwNP患者的鼻上皮细胞,real-time PCR检测miR-34c的表达,免疫细胞化学检测纤毛细胞数量。建立鼻上皮细胞气液界面培养体系,敲低表达miR-34c后免疫荧光检测纤毛标记物β-tubulinⅣ的表达。采用IL-13刺激鼻上皮细胞,real-time PCR检测miR-34c和纤毛发生转录因子FOXJ1的表达。结果与正常对照鼻上皮细胞相比,CRSwNP鼻上皮中miR-34c表达量降低(正常对照13.43±10.31 vs CRSwNP 5.53±2.56,P<0.05),纤毛细胞百分比也有明显下调(正常对照18.20%±2.52%vs CRSwNP 12.58%±2.06%,P<0.05)。在鼻上皮细胞中敲低表达miR-34c后,纤毛细胞数量明显降低。IL-13刺激可以明显降低miR-34c(刺激前5.85±4.54 vs刺激后0.43±0.40,P<0.05)和FOXJ1(刺激前1.41±1.09 vs刺激后0.15±0.19,P<0.05)的表达量。结论IL-13可以通过调控miR-34c表达进而调控纤毛细胞分化,从而参与CRSwNP黏液纤毛功能障碍。
OBJECTIVE To explor e the effects and regulating mechanisms of miR-34c on epithelial ciliated cell differentiation in chronic rhinosinusitis with nasal polyps(CRSwNP).METHODS Nasal epithelial cells were obtained from control subjects and CRSwNP patients,the expression of miR-34c was detected by real-time PCR,and the ciliated cell numbers were quantified by immunocytochemistry.The miR-34c inhibitor was transfected into the air-liquid interface(ALI)nasal epithelial cultures,immunofluorescence was used to detect the expression of ciliary markerβ-tubulinⅣ.In addition,real-time PCR was used to detect the expression of miR-34c and FOXJ1 mRNA after IL-13 stimulation on nasal epithelial cells.RESULTS Compared with control epithelial cells,the expression of miR-34c(control 13.43±10.31 vs CRSwNP 5.53±2.56,P<0.05)and ciliated cell percentage(control 18.20%±2.52%vs CRSwNP 12.58%±2.06%,P<0.05)were decreased in CRSwNP-derived epithelial cells.Transfection with miR-34c inhibitor induced a decrease of ciliated cell number in ALI cultures.IL-13 stimulation induced a significant decrease of both miR-34c(pre-stimulation 5.85±4.54 vs post-stimulation 0.43±0.40,P<0.05)and FOXJ1 expression(pre-stimulation 1.41±1.09 vs post-stimulation 0.15±0.19;P<0.05).CONCLUSION IL-13 played a role in regulating ciliated cell differentiation by regulating miR-34c expression,thus involved inmucociliary dysfunction in CRSwNP.
作者
矫健
李景云
李颖
张罗
JIAO Jian;LI Jingyun;LI Ying;ZHANG Luo(Department of Otolaryngology Head and Neck Surgery,Beijing Tongren Hospital,Capital Medical University,Beijing Institute of Otolaryngology,Key Laboratory of Otolaryngology Head and Neck Surgery(Capital Medical University),Ministry of Education,Beijing Key Laboratory of Nasal diseases,Beijing,100005,China)
出处
《中国耳鼻咽喉头颈外科》
CSCD
2020年第1期30-33,共4页
Chinese Archives of Otolaryngology-Head and Neck Surgery
基金
北京市自然科学基金面上项目(7172053、7182034)
国家重点研发计划(2016YFC0905200)
教育部长江学者及创新团队发展计划(IRT13082)
国家自然科学基金重点项目(81630023)
国家自然科学基金面上项目(81970849)
北京市医管局使命计划(SML20150203)
北京市卫生系统高层次卫生技术人才计划(2014-3-018)。
