烟曲霉提取物诱导小鼠哮喘模型的建立

Aspergillus fumigatus induced asthma model

目的:探讨小鼠哮喘模型建立的方法,为开展哮喘发生机制或药物治疗研究奠定基础。方法:40只,SPF级C57BL/6小鼠,雌雄各20只,随机分成对照组和模型组两组。模型组小鼠采用气管滴注烟曲霉提取物,隔天给药,对照组小鼠采用无菌0.9%氯化钠溶液气管滴注。给药21天后,最后一次给药24小时后收取肺泡灌洗液,离心后灌洗液上清检测炎症因子水平,灌洗细胞做Giemsa染色分类计数;取左肺做AB-PAS染色,右肺提取mRNA;腹主动脉取血ELISA检测Ig E水平。结果:与对照组相比,模型组小鼠可见典型的哮喘症状,肺泡灌洗液中炎症细胞总数、嗜酸性粒细胞和中性粒细胞较对照组显著增多,肺泡灌洗液IL-3、IL-4、IL-5、IL-6及IL-17表达上调,与对照组相比分别为P<0.05和P<0.01。血清Ig E较对照组明显增加[(23.84±3.65)vs.(6.22±1.06)mg/L,P<0.01];AB-PAS染色黏液分泌较对照组明显加重。结论:烟曲霉提取物气管滴注可成功诱导小鼠哮喘模型。

Objective：To investigate the method of mice asthma model, and set the foundation for mechanisms study and drug treatment. Methods： 40 SPF grade C57B6/L mice were randomly divided into control group and model group, half male and half female. Model mice were administrated with tracheal instillation of Aspergillus fumigatus （A. f. ） on another day, control mice with sterile saline. Challenged for 21 days, at 24 h after the final challenge, collected the total Bronchoalveolar lavage fluid （BALF）. The BALF was centrifuged and the supernatant cytokines was quantification. The BALF cell analyzed for differential cell counts performed after Wright-Giemsa staining. The left lobe of the lung was prepared for AB-PAS staining and the fight lobe of the lung used for RNA isolation. Abdominal aorta serum IgE test by ELISA. Results： Compared with saline treatment group, model mice showed typical symptoms of asthma. The total inflammatory cells, eosinophils and neutrophils in BALF were significantly increased compared with the control group. BALF IL-3, IL-4, IL-5, IL- 6 and IL-17 expression were increased compare with the control group, P 〈 0. 05 and P 〈0. 01, repectively. Serum IgE was significantly increased compared with the control group [ （ 23.84± 3.65 ） vs. （ 6. 22 ± 1.06 ） mg/L, P 〈 0. 01] ; AB-PAS staining mucus secretion significantly increased than the control group. Conclusion： Aspergillus fumigatus administrated tracheal instillation can cause mice asthma model.