miR-205-5p靶向HMGB1/TLR4途径对变应性鼻炎免疫功能的作用研究  

Effect of miR-205-5p targeting HMGB1/TLR4 pathway on immune function in allergic rhinitis

作　　者：匡玉婷[1] 胡彬雅[1] 赵斯君[1] 黄敏[1] 吴雄辉[1] 龙松良[1] 张梦萍 郑立春 KUANG Yuting;HU Binya;ZHAO Sijun;HUANG Min;WU Xionghui;LONG Songliang;ZHANG Mengping;ZHENG Lichun(Department of Otolaryngology Head and Neck Surgery,Hunan Provincial Children’s Hospital,Changsha 410007,China)

机构地区：[1]湖南省儿童医院耳鼻咽喉头颈外科,湖南长沙410007

出　　处：《中国耳鼻咽喉颅底外科杂志》2022年第4期57-62,共6页Chinese Journal of Otorhinolaryngology-skull Base Surgery

基　　金：湖南省自然科学基金科卫联合项目(2021JJ70079);湖南省自然科学基金科卫联合项目(2021JJ70078)。

摘　　要：目的研究微小RNA-205-5p(miR-205-5p)靶向高迁移率族蛋白B1/Toll样受体4(HMGB1/TLR4)途径改善变应性鼻炎(AR)免疫功能的作用。方法选取60只雄性大鼠,分为正常对照组(A组)、变应性鼻炎组(B组)、变应性鼻炎+miR-205-5p agomir组(C组)、变应性鼻炎+miR-205-5p antagomir组(D组)共4组,每组各15只,A组不做任何处理,B、C、D组均采用卵清白蛋白鼻腔强化致敏建立AR模型。模型建立完成后,C组尾部注射300μg miR-205-5p agomir,D组尾部注射miR-205-5p antagomir,A、B组均注射生理盐水。1周后评价各组行为学、免疫相关指标[干扰素-γ(IFN-γ)、白介素-2(IL-2)、白介素-4(IL-4)、白介素-10(IL-10)、免疫球蛋白E(Ig E)]、鼻黏膜炎性反应、上皮细胞间紧密连接、内质网形态及紧密连接蛋白、HMGB1/TLR4途径蛋白表达量。miR-205-5p与TLR4的靶向关系使用双荧光素酶报告实验分析。结果4组行为学评分、IL-4、Ig E、HMGB1、TLR4、NF-κB表达量、紧密连接宽度从低到高分别为A组<B组<C组<D组;IFN-γ、IL-2、IL-10、紧密连接蛋白表达量从低到高分别为D组<C组<B组<A组,且各组之间比较,差异均具有统计学意义(P均<0.05)。双荧光素酶报告结果表明miR-205-5p可与TLR4靶向结合。结论miR-205-5p可靶向HMGB1/TLR4途径抑制Th1/Th2失衡所引起的AR免疫障碍,加强鼻黏膜屏障功能,从而减少变应原的入侵,缓解AR相关症状,对AR的治疗产生积极作用。Objective To study the effect of microrna-205-5p(mir-205-5p)targeting high mobility group protein B1/toll like receptor 4(HMGB1/TLR4)pathway on immune function of allergic rhinitis(AR).Methods Sixty male rats were selected and divided into normal control group(group A),AR group(group B),AR+miR-205-5p agomir group(group C),AR+miR-205-5p antagomir group(group D)with 15 rats in each group.The rats of group A did not receive any treatment,while those of group B,C,and D were sensitized with ovalbumin nasal enhancement sensitization to establish AR model.After model establishment,300μg of miR-205-5p agomir and miR-205-5p antagomir were injected into the tail of rats of group C and group D respectively,and normal saline into both group A and group B.One week later,the behavioral,immune-related indexes[including interferon-γ(IFN-γ),interleukin-2(IL-2),interleukin-4(IL-4),interleukin-10(IL-10),immunoglobulin E(Ig E)],nasal mucosal inflammatory response,tight junctions between epithelial cells,endoplasmic reticulum morphology and expressions of tight junction proteins,HMGB1/TLR4 pathway proteins were evaluated.The targeting relationship between miR-205-5p and TLR4 was analyzed using a dual-luciferase reporter assay.Results The orders from lowest to highest of behavioral score,expression levels of IL-4,Ig E,HMGB1,TLR4,and NF-κB as well as tight junction width were group A,group B,group C and group D.The orders from lowest to highest of expression levels of IFN-γ,IL-2,IL-10 and tight junction protein were group D,group C,group B and group A,and their differences among different groups were all statistically significant(all P<0.05).The dual-luciferase reporter results indicated that miR-205-5p could target TLR4.Conclusion miR-205-5p can target the HMGB1/TLR4 pathway to inhibit the AR immune disorder caused by Th1/Th2 imbalance,strengthen the barrier function of the nasal mucosa,thereby reducing the invasion of allergens,relieving AR-related symptoms,and having a positive effect on the treatment of AR.

关 键 词：微小RNA-205-5p 高迁移率族蛋白B1/Toll样受体4 变应性鼻炎 免疫功能 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]