Notch1-Jagged1信号在变应性鼻炎中的表达及意义

摘要

目的 探讨Notch1-Jagged1信号在变应性鼻炎(AR)模型小鼠各发病阶段鼻黏膜及AR患者血清中的表达及意义.方法 将36只小鼠分为正常对照组、卵清蛋白(OVA)基础致敏组及OVA/AR组,每组12只.造模后对各组小鼠进行AR症状评分,苏木精-伊红(HE)染色法观察小鼠鼻黏膜中嗜酸粒细胞浸润情况;酶联免疫吸附试验(ELISA)法检测外周血中总IgE水平;流式细胞术检测脾脏中调节性T细胞(Treg细胞)比例变化;Western blot法检测鼻黏膜中Notch1、Jagged1的表达水平;多重液相蛋白定量技术(cytometric bead array,CBA)检测脾脏淋巴细胞中Th1/Th2/Th17相关细胞因子水平.收集2017年6-10月期间就诊于武汉大学人民医院耳鼻咽喉头颈外科的50例AR患者和30例对照组人群的血清,ELISA法检测血清中Notch1、Jagged1的表达情况.多组间比较采用单因素方差分析,两组间比较采用LSDt检验;应用Pearson相关分析评价Notch1与各细胞因子相关性.结果 与正常对照组比较,基础致敏组小鼠的症状、鼻黏膜嗜酸粒细胞浸润情况及血清总IgE水平的差异无统计学意义;OVA/AR组小鼠的症状、鼻黏膜嗜酸粒细胞浸润情况及血清总IgE水平明显升高[(6.11±0.78)分((x)±s,下同)比(0.67±0.50)分,(77.67 ±5.61)个比(10.33±0.82)个,(106.80±11.91) pg/ml比(82.45±19.80) pg/ml,t值分别为19.471、34.848、2.542,P值均＜0.05].与正常对照组比较,基础致敏组小鼠脾脏中Treg细胞比例上升,OVA/AR组Treg细胞比例下降[(10.29±0.47)％比(9.28±0.16)％,(8.49±0.15)％比(9.28±0.16)％,t值分别为5.838、4.540,P值均＜0.01].与正常对照组比较,基础致敏组与OVA/AR组小鼠鼻黏膜中的Notch1表达均上升(1.04±0.05比0.71 ±0.05,1.83 ±0.10比0.71 ±0.05,t值分别为9.293、31.363,P值均＜0.01);OVA/AR组Jagged1表达上升(0.41±0.04比0.21±0.01,t=13.472,P＜0.01).相关分析发现Notch1与IL-6、IL-10表达呈正相关(r值分别为0.98、0.87,P值均＜0.01).与对照组比较,AR组患者血清的Notch1、Jagged1表达明显上升[(1 135.0 ±254.9)pg/ml比(436.0±139.3)pg/ml,(1 200.2±401.0) pg/ml比(559.9±124.2)pg/ml,t值分别为13.99、11.94,P值均＜0.01].结论 Notch1受体与配体在AR发病过程中高表达,Notch1-Jagged1信号轴通过上调IL-6、IL-10的表达促进了AR的发生、发展.%Objective To study the expression and significance of Notch1-Jagged1 in nasal mucosa of allergic rhinitis (AR) mouse model in various stages and in the serum of AR patients.Methods Thirty-six mice were divided into 3 groups:control group,basal sensitization group (OVA) and OVA/AR group,with 12 mice in each group.Allergic symptom in each group were scored after AR model establishment.HE staining method was used to observe the nasal mucosa eosinophils infiltration.ELISA was used to detect the serum level of total IgE.Flow cytometry was used to detect the change of Treg cells in each group.Western blot was used to detect the expression of Notch1 and Jagged1 in nasal mucosa.Flow cytometric bead array (CBA) was used to detect the level of Th1/Th2/Th17 cytokines in splenic lymphocytes.The serum was obtained from 50 patients with AR and 30 control volunteers in Department of Otorhinolaryngology Head and Neck Surgery,Renmin Hospital of Wuhan University from June to October 2017.ELISA was used to detect the expression of Notch1 and Jaggedl.Results Compared with the control group,the allergy symptom,the number of nasal mucosal eosinophils and the level of total IgE were not significantly different in basal sensitization group,but increased significantly in OVA/AR group (6.11 ± 0.78 vs 0.67 ± 0.50,77.67 ± 5.61 vs 10.33 ± 0.82,(106.80 ± 11.91) pg/ml vs (82.45 ± 19.80) pg/ml,t value was 19.471,34.848,2.542,respectively,all P ＜ 0.05).Compared with the control group,the ratio of Treg cell increased in basal sensitization group but decreased in OVA/AR group ((10.29 ± 0.47)％ vs (9.28 ± 0.16) ％,(8.49 ± 0.15) ％ vs (9.28 ± 0.16) ％,t value was 5.838,4.540,respectively,all P ＜ 0.01).Compared with control group,the expression of Notch1 increased significantly both in basal sensitization group and OVA/AR group (1.04 ±0.05 vs 0.71 ±0.05,1.83 ±0.10 vs 0.71 ±0.05,t value was 9.293,31.363,respectively,all P ＜ 0.01);and the expression of Jagged1 only increased significantly in OVA/AR group (0.41 ±0.04 vs 0.21 ±0.01,t =13.472,P ＜0.01).It was found that Notch1 was positively correlated with the level of IL-6,IL-10 by Pearson test (r value was 0.98,0.87,respectively,all P ＜ 0.01).Compared with control volunteers,the expression of Notch1 and Jagged1 increased significantly in AR group patients ((1 135.0 ± 254.9) pg/ml vs (436.0 ± 139.3) pg/ml,(1 200.2 ± 401.0) pg/ml vs (559.9±124.2) pg/ml,t value was 13.99,11.94,respectively,allP＜0.01).Conclusions The expression of Notch1 receptor and ligand increased significantly in the pathogenesis of AR.Notch1-Jagged1 may promote the occurrence and development of AR by up-regulating the expression of IL-6 and IL-10.