调节性T细胞和辅助性T细胞17的免疫平衡与分泌性中耳炎动物模型的相关性研究  

Research on correlation between Treg/Th17 immune balance and animal model of otitis media with effusion

作　　者：李娟娟 唐志元 张艳红 王朝霞 唐志辉[2] 曾宪海 LI Juanjuan;TANG Zhiyuan;ZHANG Yanhong;WANG Zhaoxia;TANG Zhihui;ZENG Xianhai(Department of Otolaryngology,Shenzhen Longgang E.N.T Hospital,Shenzhen Key Laboratory of E.N.T Institute,Shenzhen,Guangdong,518172,China;Department of Otolaryngology Head and Neck Surgery,the Chinese University of Hong Kong,Hong Kong SAR,China)

机构地区：[1]深圳市龙岗区耳鼻咽喉医院耳鼻咽喉科深圳市耳鼻咽喉研究所,广东深圳518172 [2]香港中文大学医学院耳鼻咽喉头颈外科学系,中国香港

出　　处：《中国耳鼻咽喉头颈外科》2022年第7期420-424,共5页Chinese Archives of Otolaryngology-Head and Neck Surgery

基　　金：深圳市医学重点学科建设经费资助项目(SZXK039);深圳市龙岗区经济与科技发展专项资金医疗卫生科技计划项目(LGKCYLWS2019000227)。

摘　　要：目的探讨调节性T细胞和辅助性T细胞17(Treg/Th17)免疫平衡与分泌性中耳炎(otitis media with effusion,OME)动物模型中的相关性研究及初步作用机制。方法建立OME大鼠模型,采用HE染色、免疫组织化学、蛋白质免疫印迹和流式细胞术检测中耳黏膜和脾脏淋巴细胞的变化以及中耳黏膜PI3K/Akt/mTOR信号通路的变化。结果模型组(OME组)中视黄酸相关孤儿受体γt(RORγt)抗原表达阳性细胞率较对照组(CON组)强;OME组中耳黏膜中叉头状转录因子3(FOXP3)抗原表达较CON组弱;与CON组相比,OME组Treg比例为3.57%±0.66%,CON组Treg比例为14.99%±2.49%;OME组Th17比例为12.17%±1.90%,CON组Treg比例为3.29%±0.49%;与CON组相比,OME组中耳黏膜中雷帕霉素靶蛋白(m TOR)、丝氨酸/苏氨酸蛋白激酶(Akt)、磷脂酰肌醇3-激酶(PI3K)蛋白的磷酸化水平增高。结论通过对动物模型的验证,说明OME可能是全身过敏反应在中耳局部的表现,Treg/Th17表达失衡,PI3K/Akt/mTOR信号通路被激活,导致中耳黏膜发生充血、水肿和中耳积液。OBJECTIVE To explore the correlation between Treg/Th17 immune balance and otitis media with effusion in animal models and its mechanism.METHODS Rat models of otitis media with effusion were established.The changes of lymphocytes in middle ear mucosa and spleen as well as PI3 K/Akt/mTOR signaling pathway in middle ear mucosa were detected by IHC,HE,WB and flow cytometry.RESULTS The rat model of otitis media with effusion was successfully established.The expression rate of RORyT antigen positive cells in the middle ear mucosa of the model group was higher than those of the control group.The expression of FOXP3 antigen in the middle ear mucosa of the model group was lower than that of the control group.Compared with the control group,the proportion of Treg in model group was 3.57%±0.66%,and that in the control group was 14.99%±2.49%.The proportion of Th17 in model group was 12.17%±1.90%,and that of Treg in the control group was 3.29%±0.49%.Compared with the control group,the phosphorylation levels of PI3 K,mTOR and Akt were increased in the middle ear mucosa of the model group.CONGLUSION Our animal experiment suggests that secretory otitis media may be the local manifestation of systemic allergic reaction in the middle ear.The imbalance of Treg/T17 expression and activation of PI3 K/Akt/mTOR signaling pathway lead to hyperemia and edema of middle ear mucosa and middle ear effusion.

关 键 词：伴渗出液中耳炎 T淋巴细胞 调节性 T淋巴细胞 辅助诱导 辅助性T细胞17 

分 类 号：R764.21[医药卫生—耳鼻咽喉科]