C57bl/6小鼠OVA哮喘模型的建立与评价

Establishment of a C57bl/6 mouse asthma model by ovalbumin and its evaluation

摘要:

目的：本实验旨在利用无鸡蛋卵清蛋白（ovalbumin，OVA）诱导建立C57bl/6小鼠的哮喘模型，并用地塞米松对该模型小鼠进行治疗，用以评价模型是否建立成功。方法：将36只３代以上脱敏饮食的6周龄雌性C57bl/6小鼠随机分为对照组、哮喘组、地塞米松治疗组。哮喘组小鼠在第0、14天分别腹腔注射0.2 mL致敏液[20 μg OVA，100 μL生理盐水，100 μL Al（OH）3]致敏，在第21~27天用1.5% OVA溶液雾化，对照组小鼠在同样时间给予同剂量的生理盐水，治疗组在第25~27天雾化前0.5 h腹腔注射5 mg/kg地塞米松溶液0.2 mL。于末次雾化后24 h内对每组6只小鼠进行有创肺功能检测气道阻力，其余6只进行支气管肺泡灌洗观察气道炎症情况，HE染色观察肺部病理改变，PAS糖原染色观察气道黏液分泌情况，ELISA检测血清IgE以及肺泡灌洗液中IL-4含量。结果：哮喘组小鼠气道阻力值在激发浓度为25 mg/mL时较对照组（F=26.53０，P=0.000）明显升高，在激发浓度为50 mg/mL时较对照组（F=18.690，P=0.000）明显升高，支气管肺泡灌洗液（bronchoalveolar lavage fluid，BALF）细胞计数细胞总数哮喘组较对照组（F=1.349，P=0.000）和治疗组（F=1.461，P=0.006）有明显增多，嗜酸性粒细胞分类计数哮喘组较对照组（F=7.000，P=0.013）和治疗组相比也有增多，但差异无统计学意义（F=1.000，P=0.056），肺部病理和气道黏液分泌情况哮喘组与对照组相比均较严重，哮喘组血清中IgE含量与对照组相比有统计学差异（F=15.10，P=0.036）和治疗组相比有所增加，但差异无统计学意义（F=2.680，P=0.083），哮喘组肺泡灌洗液中IL-4含量均较对照组（F=7.953，P=0.000）和治疗组（F=4.413，P=0.008）明显增加。结论：本实验成功确立了构建C57bl/6小鼠哮喘稳定模型的方法。

Abstract:

Objective：To establish a C57bl/6 mouse asthma model the ovalbumin（OVA） free and to evaluate the model using dexam－ethasone（Dex）. Methods：Clean female C57bl/6 mice fed without OVA for over 3 generations were divided into three groups（n=36，12 for each group）. The asthmatic group was sensitized by i.p with OVA aluminium hydroxide gel on day 1 and 14. From 21st day，asthmatic group was aerosolized 1.5% OVA for 7 days. The control group received saline as the substitution of OVA. The treated group was i.p with 5 mg/kg dexamethasone 0.5 h before atomization from 25th to 27th day. Twenty four hour after last atomization，6 mice in each group were used to detect the airway resistance by invasive pulmonary function. The last 6 mice were used to detect airway in－flammatory cells，lung pathological changes，mucus hypersecretion，total IgE in serum and IL-4 by bronchoalveolar lavage fluid（BALF），hematoxylin-eosin（HE） staining，periodic acid-schiff（PAS） staining and ELISA. Results：The airway resistance lung resistant in asthma group was higher than that in control group at 25 mg/mL concentration（F=26.530，P=0.000） and also higher than that in control group at 50 mg/mL concentration（F=18.690，P=0.000）. BALF total cells of asthma group were higher than that in control group（F=1.349，P=0.000） and treatment group（F=1.461，P=0.006）. The eosinophils cells of asthma group was higher than that of control group（F=7.000，P=0.013）. Lung pathological changes and mucus hypersecretion in asthma group were much serious. Total IgE in serum（F=15.10，P=0.036） and IL-4 in BAlF（F=15.10，P=0.036） of asthma group were more serious than those of control group. Conclusion：The C57bl/6 mouse allergic asthma model is es－tablished by the way in this study.