屋尘螨诱导过敏性哮喘伴认知功能障碍小鼠模型的构建

Construction of a mouse model of house dust mite-induced allergic asthma with cognitive impairment

摘要

目的 旨在构建屋尘螨诱导过敏性哮喘伴认知功能障碍的小鼠模型。 方法 以6～8周龄C57 BL/6小鼠为研究对象，采用屋尘螨（house dust mite，HDM）滴鼻诱导小鼠产生过敏性哮喘（HDM组），同时设立生理盐水对照组（normal saline control group，NS组）和空白对照组（blank control group，CN组），并分别在构建模型的2周和1个月，通过巴恩斯迷宫检测认知能力，乙酰-β-甲基氯化胆碱溶液雾化检测肺功能，苏木素-伊红（hematoxylin-eosin，HE）染色、免疫组化和免疫印迹检测肺和脑组织病理变化等对模型进行评估。 结果 与2周CN组和NS组比较，HDM组小鼠增强呼吸间歇（enhanced pause，Penh）和支气管周围炎性细胞浸润差异无统计学意义；小鼠找到逃生箱的时间和次数差异无统计学意义；脑内神经元标志物成熟神经元相关神经元核（neuronal nuclei antigen，NeuN）和突触蛋白变化无明显差异；脑内未出现神经炎症。而与1个月CN组和NS组比较，HDM组小鼠Penh出现明显升高，支气管周围出现大量炎性细胞浸润；小鼠找到逃生箱的时间明显增加（F=19.600，P<0.001），找到逃生箱的次数明显减少（F=10.150，P=0.002）；小鼠脑内海马区NeuN阳性面积明显减少（F=6.449，P=0.012），突触蛋白明显减少（F=16.200，P=0.004）；出现小胶质细胞活化和神经炎症。 结论 成功构建的屋尘螨诱导过敏性哮喘伴认知功能障碍小鼠模型，为研究过敏性哮喘引起认知功能障碍的机制探讨及药物干预研究提供了一种较好的动物模型。

Abstract

Objective To construct a mouse model of house dust mite（HDM）-induced allergic asthma with cognitive impairment. Methods C57 BL/6 mice aged 6-8 weeks were used as the study subjects. HDMs were used intranasally to induce allergic asthma in mice（HDM group）. Simultaneously，a normal saline control group（NS group） and a blank control group（CN group） were established. At week 2 and month 1 of model construction，cognitive ability was tested using Barnes maze；pulmonary function was determined by nebulization with acetyl-β-methylcholine chloride solution，and pathological changes in lung and brain tissues were evaluated using hematoxylin-eosin staining，immunohistochemistry，and Western blotting. Results At week 2，there were no significant differences in the enhanced pause（Penh），inflammatory cell infiltration around the bronchi，time and frequency of mice finding escape boxes，and changes in mature neuronal marker neuronal nuclei antigen（NeuN） and synaptophysin in the brain in the HDM group compared with the CN and NS groups；no neuroinflammation occurred in the brain. At month 1，compared with the CN and NS groups，the HDM group showed a significant increase in Penh and a large amount of inflammatory cell infiltration around the bronchi；the time of mice finding escape boxes significantly increased（F=19.600，P<0.001），and the frequency significantly decreased（F=10.150，P=0.002）；the NeuN-positive area in the hippocampus of mice significantly decreased（F=6.449，P=0.012），and synaptophysin was significantly reduced（F=16.200，P=0.004）；activation of microglial cells and neuroinflammation occurred. Conclusion This study successfully constructed a mouse model of HDM-induced allergic asthma with cognitive impairment，providing a good animal model for exploring the mechanism of cognitive impairment caused by allergic asthma and drug intervention research.

关键词

过敏性哮喘 / 认知功能障碍 / 屋尘螨 / 动物模型 / 小鼠

Key words

allergic asthma / cognitive impairment / house dust mite / animal model / mouse