尘螨过敏性哮喘小鼠模型的建立与评估

Preparation and evaluation of mouse model of house dust mite induced asthma

摘要:

目的支气管哮喘（哮喘）的动物模型是研究人类哮喘的重要手段之一，但模型很难模拟人类哮喘的主要特点，而且因致敏原的不同影响了实验研究的意义。该文采用哮喘的常见致敏原尘螨抗原提取物（Derp）制作尘螨过敏性哮喘模型，并描述其气道炎症，评估其气道高反应性，尽可能与人类哮喘的病因及病变过程相似，以利于哮喘的实验研究。方法C57BL/6J小鼠分为对照组和模型组,各6只，Derp两次腹腔注射致敏(day0,day10)，麻醉后滴鼻激发隔日一次，共7次，空白对照组用生理盐水代替，末次激发后24 h检测气道高反应性(AHR)，之后进行支气管肺泡灌洗，肺组织做病理检测 (常规及特殊染色)。结果模型组气道阻力增加幅度（Raw%）、动态肺顺应性下降幅度（Cdyn％）均增高（P<0.01），Raw增加25%时(PC25Raw)，Cdyn下降15%时(PC15Cdyn)所需的MCh浓度均下降（P<0.01）；肺泡灌洗液（BALF）中细胞总数、嗜酸细胞绝对值计数以及嗜酸细胞百分比均增加（P<0.01）；模型组肺组织病理积分高于对照组（P<0.01），并且出现超微结构的改变，还可见杯状细胞，粘液及大量肥大细胞颗粒。结论 采用腹腔注射致敏并滴鼻激发的方法可以建立尘螨过敏性哮喘模型，具备了气道炎症及气道高反应性的特点。

Abstract:

ObjectiveTo prepare a mouse model of asthma by sensitizing and challenging with house dust mite allergen Derp and evaluate its reliability by measuring airway allergy inflammation and airway responsiveness. MethodsTwelve C57BL/6J mice were randomly divided into two groups: control and asthma model. Mice of the asthma model group were sensitized by intraperitoneal injection of house dust mite allergen Derp on the first and tenth days of the experiment. From the 17th day, the mice were challenged by intranasal Derp, once every other day, seven times. The control group was treated with normal sodium instead of Derp. Twentyfour hours after the last challenge, airway responsiveness was evaluated. Bronchoalveolar lavage and histological examination of the lung were performed. ResultsAirway resistance increased and dynamic lung compliance decreased significantly in the asthma model group as compared to the control group (P<0.01). When airway resistance increased by 25% and dynamic lung compliance decreased by 15%, the required metacholine concentration in the asthma model group was significantly lower than that in the control group (P<0.01). In the bronchoalveolar lavage fluid of the asthma model group, the number of total cells, absolute number of eosinophils (EOS) and the percentage of EOS in the total cell were significantly higher than those in the control group (P<0.01). Pulmonary pathological scores in the asthma model group were significantly higher than those in the control group (P<0.01). The asthma model group showed ultrastructural changes of bronchial and pulmonary arterioles. Goblet cells, mastocyte granules, and increased mucus were observed in the lung tissues of the asthma model group. ConclusionsA mouse model of asthma was prepared by sensitizing and challenging with house dust mite allergen Derp, with the characteristics of airway allergy inflammation and airway hypersensitivity reaction.