滴鼻和雾化两种不同激发方式对小鼠支气管哮喘模型气道炎症的影响

Effect of Intranasal or Aerosol Challenge on Airway Inflammation in a Murine Asthma Model

目的探讨滴鼻和雾化两种不同激发方式对小鼠支气管哮喘模型气道炎症的影响,为研究和优化支气管哮喘动物模型提供实验和理论依据。方法选择鸡卵清蛋白(OVA)致敏的BALB/c小鼠,分别采用滴鼻和雾化两种不同方式进行激发,复制哮喘模型,观察支气管肺泡灌洗液(BALF)中白细胞总数和细胞分类计数,ELISA方法检测BALF上清中IL-4、IL-13含量,常规病理切片苏木精-伊红(HE)染色观察肺内炎症情况。结果滴鼻组小鼠BALF中白细胞总数最高,其次为雾化组小鼠;且滴鼻组小鼠嗜酸性粒细胞总数及中性粒细胞总数亦明显高于雾化组与正常组,差异有统计学意义(P<0.05)。滴鼻组小鼠BALF中IL-4、IL-13水平明显高于雾化组与正常组小鼠,差异有统计学意义(P<0.05);雾化组小鼠BALF中IL-4、IL-13水平高于正常组小鼠,差异有统计学意义(P<0.05)。滴鼻组肺组织炎症反应最明显,并可见上皮细胞破坏,杯状细胞增生,支气管和血管周围有大量炎症细胞浸润,以嗜酸性粒细胞和淋巴细胞为主。雾化组炎症程度较轻,正常对照组无明显炎症改变。结论雾化吸入和经鼻滴入的激发方式都可以成功制作出支气管哮喘模型,经鼻滴入方式可以引起肺部更加明显的炎症浸润及细胞因子表达,为哮喘研究提供更好的选择。

Objective To investigate the effect of two different methods of challenge,intranasal or aerosol exposure,on air- way inflammation in murine asthma models, and provide experimental and theoretical basis for the study and optimization of bronchial asthma in animal models. Methods Ovalbumin（OVA）-sensitive BALB/e mice were challenged to set up murine mod- els of atopic asthma by intranasal or aerosol exposure to OVA. The number of leukocytes and cell classifications in bronchoalve- olar lavage fluid （BALF） were measured. The concentrations of IL-4 and IL-13 in the supernatant of BALF were determined by ELISA and the histological changes in the lung by hematoxylin and eosin （HE） staining. Results The total leukocytes were highest in the intranasal group. The number of eosinophils and neutrophils in the intranasal group were higher than that in the aerosol group and normal group （P〈0. 05）. The levels of IL-4 and IL-13 were significantly increased in the s upernatant of BALF in the intranasal group when compared with the aerosol group and normal group （P〈0.05）. The levels of IL-4 and IL-13 in the aerosol group were higher than those in the normal group （P〈0.05）. The lung tissue inflammation presented destruction of epithelial cells, hyperplasia of goblet cells and infiltration of large numbers of inflammatory cells （predominantly eosinophils and lymphocytes）. It was most obvious in the intranasal group and no obvious inflammation was observed in the normal group. Conclusion Asthmatic models can be established successfully by both of intranasal and aerosol challenge and the in- tranasal administration triggers more serious airway inflammation and higher cytokines expression than aerosol challenge, provi- ding better choice for the study of asthma.