小鼠上下气道一致的变应性气道炎症动物模型的建立

Establishment of an animal model of unified allergic airway inflammation in upper and lower airways of mouse

目的 建立小鼠上下气道一致的变应性气道炎症模型。方法 16只雌性Balb/c小鼠随机分为模型组(A组)和对照组(B组),每组各8只。A组以40μg卵白蛋白(ovalbumin, OVA)+200 mgAl(OH)_(3)+200μLPBS溶液,分别于第1、3、5、7、9、11、13天腹腔注射,共7次,从第20天起以10μL OVA(1 mg/mL)滴鼻激发,每周3次,连续3周,最后1次滴鼻激发后24 h以2%OVA 5 mL雾化吸入激发,连续5 d。B组以0.9%生理盐水代替OVA。整个造模周期为42 d。根据黏膜纤毛缺失程度,将上皮细胞状态分为4级(0~3级),末次激发后评价小鼠模型的症状、特异性IgE浓度及鼻肺黏膜的病理改变。结果 A组鼻、肺部症状评分与B组相比,差异具有统计学意义(P<0.01)。A组鼻、肺部黏膜破坏程度(1、2级)与B组相比,差异均具有统计学意义(P<0.01),而3级均无统计学意义(P>0.05)。A组鼻、肺灌洗及血清中OVA sIgE浓度均较B组明显增高(P<0.01)。A组鼻、肺部嗜酸粒细胞和杯状细胞均较B组明显增高(P<0.01)。A组鼻、肺嗜酸粒细胞数目呈显著正相关(r=0.775,P<0.01),杯状细胞数也呈正相关(r=0.723,P<0.05)。结论 本造模方法能够成功建立小鼠症状学、免疫学及病理学3方面上下气道一致的AAI模型。

Objective To establish an animal model of unified allergic airway inflammation(AAI)in upper and lower airways of mouse. Methods A total of 16 female BALB/c mice were randomly divided into model group(group A) and control group(group B), with 8 mice in each group. The mice of group A were intraperitoneally injected with 40 ug ovalbumin(OVA) + 200 mg Al(OH)_(3)+ 200 μL PBS solution on the 1 st, 3 rd, 5 th, 7 th, 9 th, 11 th, and 13 th days, with a total of 7 times. Starting from the 20 th day, the mice were challenged with 10 μL OVA(1 mg/mL) intranasally, 3 times a week, for 3 consecutive weeks, and 24 h after the last intranasal challenge, with 2% OVA 5 mL aerosol inhalation for 5 consecutive days. In group B, normal saline was used instead of OVA and the process was the same as that of group A. The whole modeling cycle lasted 42 d. After the last challenge, the symptoms, specific immunoglobulin E(sIgE) concentrations and pathological changes of nasal and pulmonary mucosae in both groups were evaluated and compared. Results The nasal and pulmonary symptom scores of group A were significantly higher than those of group B(P<0.01). The differences of nasal and pulmonary mucosal damage between the two groups at grades 1, 2 were statistically significant(P<0.01), but insignificant at grade 3(P>0.05). OVA sIgE concentrations of nasal lavage, lung lavage and serum in group A were significantly higher than those in group B(P<0.01). The cell counts of nasal and pulmonary eosinophils(EOS) and goblet cell(GC) in group A were significantly higher than those in group B(P<0.01). In group A, the cell numbers of nasal EOS and GC were positively correlated with the pulmonary ones(r=0.775, P<0.01;r=0.723, P<0.05), respectively. Conclusion This method can successfully establish an AAI model with upper and lower airway consistency in three aspects of symptom, immunology and pathology in mice.