苯并(a)芘对变应性鼻炎模型鼻黏膜功能及Treg细胞甲基化水平的影响  

Effects of benzo(a)pyrene on nasal mucosa function and methylation level of Treg cells in allergic rhinitis(AR)model

作　　者：陈国辉 刘建 孙青 秦雪梅 郭志强 CHEN Guohui;LIU Jian;SUN Qing;Qin Xuemei;GUO Zhiqiang(Department of Otolaryngology,Qingpu Branch,Zhongshan Hospital,Fudan University,Shanghai 201700,China)

机构地区：[1]复旦大学附属中山医院青浦分院耳鼻咽喉科,上海201700

出　　处：《新疆医科大学学报》2024年第8期1073-1079,共7页Journal of Xinjiang Medical University

基　　金：上海市青浦区卫生健康委员会科研课题项目(w2021-03)。

摘　　要：目的探讨苯并(a)芘对变应性鼻炎(AR)模型鼻黏膜功能及Treg细胞甲基化水平的影响。方法36只SPF级BALB/c雌性小鼠中选取12只作为正常对照组,剩余小鼠采用随机数表法分为AR模型组和苯并(a)芘干预组,各12只。AR模型组和苯并(a)芘干预组进行建模处理。建模后,苯并(a)芘干预组腹腔注射7.89 mg/kg苯并(a)芘,正常对照组和AR模型组给予等量的无菌PBS溶液腹腔注射。对各组小鼠的鼻炎症状评分和鼻腔灌洗液细胞涂片嗜酸细胞(EOS)计数、血清中卵清蛋白特异性IgE(OVA-sIgE)、粒细胞巨噬细胞集落刺激因子(GM-CSF)、肿瘤坏死因子-α(TNF-α)、白细胞介素13(IL-13)和嗜酸性粒细胞趋化因子(Eotaxin)水平、小鼠鼻黏膜病理学变化、小鼠鼻黏膜Th亚群及Foxp3甲基化和蛋白水平进行对比。结果与AR模型组相比,苯并(a)芘干预组中小鼠鼻炎症状评分和鼻腔灌洗液细胞涂片EOS计数增加最为明显(P<0.05),小鼠血清中OVA-sIgE、GM-CSF、TNF-α、IL-13和Eotaxin水平升高最为明显(P<0.05);苯并(a)芘干预后,可见大量的炎性细胞浸润,鼻黏膜上皮组织排列紊乱,增生和血管扩张严重,纤维上皮也受到严重的损伤,出现大量的杯形细胞,苯并(a)芘干预组小鼠鼻黏膜中AhR阳性细胞数量升高最为明显(P<0.05);Th1和Treg占比降低最为明显,Th2、Th17占比升高最为显著(P<0.05);小鼠脾脏CD4+CD25+调节性T细胞Foxp3基因甲基化水平升高最为明显(P<0.05),小鼠脾脏中Foxp3蛋白水平降低最为明显(P<0.05)。结论苯并(a)芘通过增加CD4+CD25+调节性T细胞Foxp3基因甲基化,降低Treg细胞比例,加重了鼻黏膜炎症和免疫失衡,进而参与变应性鼻炎的发病机制。Objective:To investigate the effects of benzo(a)pyrene on nasal mucosa function and methylation level of Treg cells in models of allergic rhinitis(AR).Methods:12 of 36 SPF-grade BALB/c female mice were selected as normal control group,and the remaining mice were randomly divided into AR model group and the benzo(a)pyrene intervention group using a random number table,with 12 mice in each group.The AR model group and the benzo(a)pyrene intervention group were subjected to modeling treatment.After the modeling,the benzo(a)pyrene intervention group was injected with 7.89 mg/kg body weight of benzo(a)pyrene.The control group and AR model group were given the same amount of sterile PBS solution intraperitoneal injection.The scores of rhinitis symptoms,EOS count in nasal perfusion fluid,serum levels of OVA-sIgE,GM-CSF,TNF-α,IL-13 and Eotaxin,pathological changes of mouse nasal mucosa,Th subgroup of mouse nasal mucosa and Foxp3 methylation and protein levels were compared in each group.Results:Compared with AR model group,the rhinitis symptom score and EOS count of nasal lavage fluid were significantly increased in the benzo(a)pyrene intervention group(P<0.05),and the serum levels of OVA-sIgE,GM-CSF,TNF-α,IL-13 and Eotaxin were significantly increased(P<0.05).After intervention of benzo(a)pyrene,a large number of inflammatory cells were infiltrated,the nasal epithelium was disordered,hyperplasia and vasodilation were serious,the fibrous epithelium was also seriously damaged,and a large number of goblet cells appeared.The number of AhR positive cells in the nasal mucosa of benzo(a)pyrene intervention group was the most significantly increased(P<0.05).The proportion of Th1 and Treg were decreased significantly,and the proportion of Th2 and Th17 were increased significantly(P<0.05)in the benzo(a)pyrene intervention group.The methylation level of Foxp3 gene in CD4+CD25+regulatory T cells in mouse spleen was increased significantly(P<0.05),and the Foxp3 protein level in mouse spleen was decreased significantly(P<0.05)in th

关 键 词：变应性鼻炎 苯并(A)芘 TREG细胞 Foxp3基因甲基化 

分 类 号：R765[医药卫生—耳鼻咽喉科]