杨树花粉过敏BALB/c小鼠动物模型的建立

BALB/c mice model of allergic diseases induced by populus pollen

目的:建立杨树花粉粗提物致敏激发的小鼠过敏模型。方法:将60只BALB/c小鼠随机均分为正常组、卵清蛋白(OVA)阳性对照组和模型组,每隔5 d经腹腔注射致敏1次,共4次,末次致敏后第5天滴鼻激发。观察支气管肺泡灌洗液(BALF)中炎症细胞变化情况;PAS染色观察气道黏液分泌情况;HE染色观察鼻黏膜、肺炎症情况;免疫组化法检测肺组织中IL-4及IFN-γ阳性细胞平均光密度值;酶联免疫吸附试验检测血清中的总IgE水平。结果:与正常组比较,模型组诱导鼻黏膜、肺组织出现明显的变应性炎症且气道黏液分泌明显增加;BALF涂片显示明显的炎症细胞(嗜酸性粒细胞、中性粒细胞)增多;肺组织中IL-4阳性细胞平均光密度值均高于正常组,IFN-γ阳性细胞平均光密度值均低于正常组;血清中总IgE高于正常组。结论:杨树花粉粗提物能够成功建立小鼠过敏模型。该模型的建立有利于过敏性疾病机制的研究。

Objective： To study performed of develop a mice model of allergic diseases induced by crude extractings from populus pollen. Methods： A total of 60 BALB/c mice were divided randomly into there groups： normal control group,Albumin Egg（ OVA）group and populus pollen model group with 20 in each. Mices were repeatedly sensitized by intraperitoneal injections of OVA or crude populus pollen extract every 5 d for four doses. Five days after the last sensitization,mices were repeatedly challenged by once daily antigen from 21-25 d. The changes of inflammatory cells in bronchoalveolar lavage fluid（ BALF） were stained with hematoxylin and eosin（ HE） to evaluate the degree of allergic inflammation. PAS staining was used to observe the secretion of airway mucus; The changes of the nasal mucosas and lungs of mice were stained with HE to evaluate the degree of allergic inflammation. And the average optical density of IL-4 and IFN-γ positive cells in lung tissue was measured by immunohistochemistry. The total Ig E in the serum was also measured by enzyme-linked immunoassay（ ELISA）. Results： Compared with the mice in normal control group,those in OVA group and model group developed obvious allergic inflammation in the nasal mucosas and lungs,and increased airway mucus secretion. The number of inflammatory cells including eosinophil and neutrophils markedly increased in BALF smear. The average optical density of IL-4 positive cells in lung tissue was all increased in OVA group and model group compared with those in normal control group,and the average optical density of IFN-γ in lung tissue was on the contrary. The total Ig E in the serum were all increased in OVA group and model group compared with those in normal control group,and the IFN-γ in the serum was significantly reduced in OVA group and model group compared with those in normal control group. Conclusion： Taken together,crude populus pollen extract can successfully induce a mice model of allergic diseases. This model is a useful tool in studying the mechanisms of allergic disease.