Abstract: Background　Glucocorticoids are commonly used in the treatment of combined allergic rhinitis and asthma syndrome（ACAD），while airway remodeling in chronic ACAD may affect their efficacy.Therefore，it is necessary to explore a new immunotherapy，which can produce synergistic effects when being used in combination with glucocorticoids.Previous studies have shown that CpG oligodeoxynucleotides（CpG-ODN） can inhibit airway inflammation in ACAD，but their inhibitory effects on airway remodeling remain unclear.Objective　To investigate the effects of CpG-ODN on airway remodeling in mice models of chronic ACAD.Methods　This study was carried out from April 2016 to March 2018．In this study，30 clean female BALB/c mice were randomly divided into the groups of Control，ACAD，ACAD/CpG-ODN，ACAD/Budesondine（BUD），and ACAD/CpG-ODN+BUD.ACAD models were made by systemic sensitization and repeated challenge with ovalbumin（OVA） or phosphate buffer saline（PBS） intranasally for 3 successive weeks，followed by aerosolly for 6 weeks.Then，a nasal instillation of CpG-ODN，BUD and CpG-ODN+BUD was given to groups of ACAD/CpG-ODN，CAAD/BUD，and ACAD/CpG-ODN+BUD，respectively，and PBS was administered intranasally to other groups.Within 10 minutes after each nasal or airway stimulation，the numbers of sneezings and scratching the nose due to itch were counted.Pathologic analyses of nasal tissues and lung sections were performed with HE and PAS staining，HE，AB-PAS and Masson's trichrome staining，respectively.The degree of airway inflammation was evaluated according to the peripheral inflammatory cell infiltration.The expression levels of transforming growth factor beta 1（TGF-β1） and alpha-smooth muscle actin（α-SMA） in lung tissues were assessed by immunohistochemical staining with the Image-Pro Plus software.Nasal symptoms were assessed directly after nasal challenge.Results　The numbers of scratching the nose and sneezings varied significantly among the groups（P<0.01）．The numbers of scratching the nose and sneezings were much more in ACAD group compared with those of the groups of control，ACAD/CpG-ODN，and ACAD/BUD and ACAD/CpG-ODN+BUD（P<0.05）．The numbers of scratching the nose and sneezings varied significantly among the groups（P<0.01）．The numbers of scratching the nose and sneezings were much more in ACAD group compared with those of other groups（P<0.05）．Histological examinations of nasal tissues stained with HE and PAS showed that the degree of alleviation of pathological changes in nasal mucosa was the greatest in ACAD/CpG-ODN group，followed by ACAD/CpG-ODN group/ACAD/BUD group，then ACAD group.Histological examinations of lung tissue sections with HE，AB-PAS and Masson's trichrome staining found that the degree of alleviation of pathological changes in lung tissues was the greatest in ACAD/CpG-ODN+BUD group，followed by ACAD/CpG-ODN group/ACAD/BUD group，then ACAD group.The scores for the degree of airway inflammation differed significantly among the groups（P<0.01）．ACAD group had higher scores in the degree of airway inflammation than every other group（P<0.05）．The expression levels of TGF-β1 and α-SMA varied obviously among the groups（P<0.01）．ACAD group had higher expression levels of TGF-β1 and α-SMA than every other group（P<0.05）．Conclusion　CpG-ODN and BUD can inhibit the occurrence of nasal inflammation and bronchopulmonary remodeling in mice models of chronic ACAD，and they may have synergistic effects in the process.

Key words: Allergic combined airway disease, Airway remodeling, CpG oligodeoxynucleotides, Budesonide, Mice, Nasal cavity, Bronchi

摘要： 背景　激素是变应性联合气道疾病（ACAD）常用的治疗方法，但慢性ACAD出现气道重塑，可能影响其疗效，故有必要寻找与激素联合并有协同作用的新型免疫治疗。前期研究发现CpG寡聚脱氧核苷酸（CpG-ODN）抑制ACAD气道炎症，但对气道重塑的效果有待研究。目的　探讨CpG-ODN对慢性ACAD小鼠模型气道重塑的影响。方法　本研究时间为2016年4月—2018年3月。30只清洁级雌性BALB/c小鼠采用随机数字表法分为正常对照组（Control组）、ACAD组、变应性联合气道疾病CpG-ODN干预组（ACAD/CpG-ODN组）、变应性联合气道疾病布地奈德干预组（ACAD/BUD组）、变应性联合气道疾病CpG-ODN+BUD联合干预组（ACAD/CpG-ODN+BUD组）。实验动物腹腔卵清蛋白（OVA）或磷酸盐缓冲液（PBS）致敏，鼻腔OVA/PBS 3周激发，随之6周雾化气道激发。ACAD/CpG-ODN组、ACAD/BUD组和ACAD/CpG-ODN+BUD组分别给予CpG-ODN、BUD和CpG-ODN+BUD滴鼻，其他组给予PBS滴鼻。每次鼻腔或气道激发后10 min内，对小鼠抓鼻和喷嚏次数进行计数。HE和PAS染色观察小鼠鼻腔组织的病理特点。HE、AB-PAS和Masson's染色观察小鼠肺组织病理变化，并行气道炎症评分。免疫组化检测肺组织转化生长因子β1（TGF-β1）和α平滑肌肌动蛋白（α-SMA），应用Image-Pro Plus图片分析软件定量评估，并进行症状评估。结果　各组小鼠抓鼻次数和喷嚏次数比较，差异均有统计学意义（P<0.01）；其中ACAD组抓鼻次数和喷嚏次数均多于Control组，ACAD/CpG-ODN组、ACAD/BUD组、ACAD/CpG-ODN+BUD组抓鼻次数和喷嚏次数均少于ACAD组（P<0.05）。HE和PAS染色显示，ACAD/CpG-ODN组和ACAD/BUD组鼻黏膜病理改变较ACAD组减轻，而ACAD/CpG-ODN+BUD组进一步减轻。HE、AB-PAS和Masson's染色显示，ACAD/CpG-ODN组和ACAD/BUD组肺组织病理改变较ACAD组减轻，而ACAD/CpG-ODN+BUD组进一步减轻。各组气道炎症评分比较，差异有统计学意义（P<0.01）；其中ACAD组气道炎症评分高于Control组，ACAD/CpG-ODN组、ACAD/BUD组、ACAD/CpG-ODN+BUD组气道炎症评分均低于ACAD组（P<0.05）。各组肺组织中TGF-β1和α-SMA表达水平比较，差异均有统计学意义（P<0.01）；其中ACAD组TGF-β1和α-SMA表达水平均高于Control组，ACAD/CpG-ODN组、ACAD/BUD组、ACAD/CpG-ODN+BUD组均低于ACAD组（P<0.05）。结论　CpG-ODN、布地奈德抑制慢性ACAD模型鼻部炎症和支气管肺组织气道重塑，两者可能具有协同作用。

关键词: 变应性联合气道疾病, 气道重塑, CpG寡聚脱氧核苷酸, 布地奈德, 小鼠, 鼻腔, 支气管