蒿属花粉过敏性哮喘小鼠模型的建立  

Establishment of mouse model of Artemisia pollen-allergic asthma

作　　者：郑卉爽 李俊达 茹一 尚腾泽 关凯 ZHENG Huishuang;LI Junda;RU Yi;SHANG Tengze;GUAN Kai(Department of Allergy,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing Key Laboratory of Precision Medicine for Diagnosis and Treatment of Allergic Diseases,National Clinical Research Center for Dermatologic and Immunologic Diseases(NCRC-DID),Beijing 100730,China)

机构地区：[1]中国医学科学院北京协和医学院北京协和医院变态(过敏)反应科,过敏性疾病精准诊疗研究北京市重点实验室,国家皮肤与免疫疾病临床医学研究中心,北京100730

出　　处：《中华临床免疫和变态反应杂志》2024年第4期376-382,共7页Chinese Journal of Allergy & Clinical Immunology

基　　金：国家自然科学基金(82070033,82370041);中国医学科学院医学与健康科技创新工程(2021-I2M-1-017)。

摘　　要：目的建立蒿属花粉致敏的过敏性哮喘小鼠模型。方法将12只无病原体雌性BALB/c小鼠随机分成对照组与模型组,模型组小鼠每周致敏1次,致敏4次后于第25~29天连续滴鼻激发5 d。在第30天时,检测小鼠气道阻力和肺部顺应性后,杀鼠取材检测血清总免疫球蛋白E(total immunoglobulin E,T-IgE)和大籽蒿特异性免疫球蛋白E(specific immunoglobulin E,sIgE)水平,检测肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中白细胞总数及白细胞分类计数、细胞因子含量,进行肺组织病理评估分析、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate poly-acrylamide gel electrophoresis,SDS-PAGE)和免疫印迹分析(western blot analysis,WB)。结果相较于对照组,模型组小鼠气道阻力升高,肺部顺应性下降;血清中T-IgE、sIgE和BALF中嗜酸性粒细胞、中性粒细胞、白细胞介素(Interleukin,IL)-4、IL-5、IL-9、IL-13的含量均显著升高。肺组织苏木精和伊红(hematoxylin-eosin staining,HE)及高碘酸希夫(periodicacid-schiff reaction,PAS)染色分析发现,模型组小鼠支气管狭窄、炎症细胞浸润、肺部结构破坏、杯状细胞增生。WB分析发现,模型组小鼠的sIgE抗体主要与分子量为9 kDa~70 kDa的蛋白质结合,蒿属花粉过敏患者血清sIgE抗体主要与分子量为28 kDa和70 kDa的蛋白质结合。结论本研究建立了蒿属花粉过敏性哮喘小鼠模型,其可用于蒿属花粉过敏性哮喘发病机制和特异性免疫治疗相关研究。Objective To establish mouse model of Artemisia pollen-allergic asthma.Methods Twelve pathogen-free female BALB/c mice were randomly divided into control and model groups.Mice in model group were sensitized once per week for 4 weeks,then nasal drops were administered daily since 25th to 29th days after last sensitization.Mice were killed and blood samples were collected for serum total immunoglobulin E(T-IgE)and Artemisia pollen specific immunoglobulin E(sIgE)detection after evaluations of airway resistance and lung compliance on 30th day.Total number of leukocytes,differential leukocyte count and cytokines levels of bronchoalveolar lavage fluid(BALF)were also detected.Finally,pathological analysis of lung tissue was performed.Allergens of Artemisia pollen were isolated and identified by using sodium dodecyl sulfate poly acrylamide gel electrophoresis(SDS-PAGE)and Western blot analysis(WB).Results Elevated airway resistance and decreased lung compliance demonstrated among mice in model group,while serum levels of T-IgE and sIgE as well as eosinophils,neutrophils,IL-4,IL-5,IL-9 and IL-13 in BALF significantly increased,compared with that of mice in control group.Bronchial stenosis,infiltration of inflammatory cells,destruction of lung structure and hyperplasia of cup cells were revealed in lung tissues of mice in model group by using Hematoxylin-eosin(HE)and periodicacid-schiff reaction(PAS)staining.Results of Western blotting showed that sIgE antibodies of mice in model group mainly bound to 9 kDa to 70 kDa proteins bands,whereas serum sIgE antibodies of Artemisia pollen-allergic patients were mainly bound to 28 kDa and 70 kDa proteins bands.Conclusion A mouse model of Artemisia pollen-allergic asthma was successfully established,which can be used for research of pathogenesis and allergen specific immunotherapy of Artemisia pollen-allergic asthma.

关 键 词：过敏性哮喘 动物模型 蒿属花粉 

分 类 号：R593.1[医药卫生—内科学] R-332[医药卫生—临床医学]