山西医科大学学报2025，Vol.56Issue(1)：57-65,9.DOI:10.13753/j.issn.1007-6611.2025.01.009
敲低MUC5B表达对分泌性中耳炎大鼠炎症反应的影响
Effect of MUC5B knockdown on inflammatory responses in rats with secretory otitis media
摘要
Abstract
Objective To investigate the effect of Mucins 5B(MUC5B)on inflammatory response in secretory otitis media(SOM)and its relationship with the formation of neutrophil extracellular traps(NETs).Methods SOM rat model was constructed by intraperito-neal injection of ovalbumin.SOM rats were divided into model group,shRNA-NC group and shRNA-MUC5B group,with 10 rats in each group,and ten healthy SD rats were selected as control group.At 24 h after the last injection,the inflammation was scored,the lesion of middle ear mucosa tissue was detected by HE staining,the expressions of inflammatory factors in middle ear irrigation solu-tion were detected by ELISA,the distribution of Th1 and Th2 cells was detected by flow cytometry,and the expressions of NET mark-ers myeloperoxidase(MPO)and citrullinated histone H3(CitH3)in middle ear mucosa were detected by immunofluorescence staining.The ROS level in middle ear mucosa tissue was detected by DCFH-DA probe.The expression level of MUC5B gene was detected by RT-qPCR,and the expressions of interferon-γ(IFN-γ),interleukin-2(IL-2),IL-4,IL-13,MUC5B,MPO,CitH3 and neutrophil elastase(NE)proteins were detected by Western blot.Results Compared with control group,the middle ear mucosa was thickened,the in-flammation degree score was increased in model group and shRNA-NC group(P<0.05),the mRNA level of MUC5B,the protein levels of CitH3,MPO,MUC5B,IL-4 and IL-13,Th2 cell proportion and ROS levels were all increased in middle ear mucosa tissues(P<0.05),and the protein levels of IFN-γ and IL-2,Th1 cell proportion and Th1/Th2 ratio were decreased(P<0.05);the protein levels of MPO,NE and CitH3,and the contents of IL-4 and IL-13 were increased in middle ear lavage fluid(P<0.05),while the contents of IFN-γ and IL-2,and IFN-γ/IL-4 ratio were decreased(P<0.05).Compared with shRNA-NC group,the mucosal thickness and the in-flammatory degree score were decreased in shRNA-MUC5B group(P<0.05),the mRNA level of MUC5B,the protein levels of CitH3,MPO,MUC5B,IL-4 and IL-13,Th2 cell proportion and ROS level were all decreased in middle ear mucosa tissues(P<0.05),and the protein levels of IFN-γ and IL-2,Th1 cell proportion,and Th1/Th2 ratio were increased(P<0.05);the protein levels of MPO,NE and CitH3,and the contents of IL-4 and IL-13 were decreased in middle ear lavage fluid(P<0.05),while the contents of IFN-γ and IL-2,and IFN-γ/IL-4 ratio were increased(P<0.05).Conclusion MUC5B knockdown can alleviate the inflammatory response of middle ear mucosa in SOM rats by inhibiting the formation of NETs.关键词
黏蛋白5B(MUC5B)/分泌性中耳炎/炎症反应/中性粒细胞胞外诱捕网(NETs)/中耳黏膜/Th1/Th2Key words
Mucins 5B(MUC5B)/secretory otitis media/inflammatory response/neutrophil extracellular traps(NETs)/middle ear mucosa/Th1/Th2分类
医药卫生引用本文复制引用
王丹,杨启梅,刘晓娜,陈菁华,王玉霞..敲低MUC5B表达对分泌性中耳炎大鼠炎症反应的影响[J].山西医科大学学报,2025,56(1):57-65,9.基金项目
陕西省重点研发计划项目(2023-YBSF-081) （2023-YBSF-081）
