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目的:评估不同剂量过敏原对滴鼻制备小鼠急性和亚急性哮喘模型的影响。方法:将不同剂量卵清蛋白(OVA)25μg、50μg、100μg连续滴鼻,建立急性和亚急性小鼠哮喘模型。使用有创小鼠肺功能仪测量各组小鼠肺功能参数;收集小鼠肺泡灌洗液计数细胞总数和细胞分类;取其肺组织常规石蜡包埋切片,进行伊红染色(HE)、刚果红染色、马松染色和过碘酸雪夫氏(PAS)染色,观察炎性细胞浸润,胶原纤维沉积、黏液分泌等指标的变化。结果:在实验第20d和24d,100μg与50μgOVA激发组小鼠气道反应性、肺泡灌洗液中细胞总数和嗜酸性粒细胞百分比增高;小鼠肺泡、管腔、管壁及伴行动脉周围有大量炎性细胞特别是嗜酸性粒细胞浸润,支气管粘膜皱襞增多、延长,管腔内存有大量黏液性痰栓;支气管管壁及其伴行动脉周围胶原纤维沉积明显增加;气道粘蛋白染色成强阳性反应,较25μgOVA激发组和生理盐水组小鼠有明显差异。结论:采用不同剂量OVA滴鼻,成功建立了程度不同的急性和亚急性小鼠支气管哮喘模型。过敏原剂量对模型建立有重要影响,为进一步研究哮喘的发生机制和药物筛选奠定了基础。
Objective: To evaluate the effects of different doses of allergens on acute and subacute asthma models of intranasal preparation in mice. Methods: Acute and subacute mouse asthma models were established by intranasal instillation of different doses of ovalbumin (OVA) 25μg, 50μg, 100μg. The pulmonary function parameters of mice in each group were measured by using the invasive mouse lung function instrument. The total number of alveolar lavage fluid and the cell classification were collected. The paraffin-embedded sections of the lung tissue were collected and subjected to eosin staining (HE), Congo Red staining, Matson staining and periodic acid Schiff (PAS) staining were used to observe the changes of inflammatory cell infiltration, collagen deposition and mucus secretion. Results: On the 20th day and the 24th day of the experiment, the airway responsiveness, the total number of cells in the BALF and the percentage of eosinophils increased in 100μg and 50μg OVA challenge group mice; the alveoli, lumen, A large number of inflammatory cells, especially eosinophil infiltration, bronchial mucosa fold increased, prolonged, a large number of mucus in the lumen of the sputum suppository; bronchial wall and its accompanying artery collagen fibers deposition increased significantly; airway mucin staining Chengqiang positive reaction, compared with 25μg OVA challenge group and saline group there are significant differences in mice. CONCLUSIONS: Acute and subacute mouse bronchial asthma models of different degrees have been established successfully with different dosages of OVA intranasally. Allergen dose has an important influence on model establishment, which lays the foundation for further research on the mechanism of asthma and drug screening.