互隔交链孢霉诱导小鼠气道变态反应性炎症动物模型的构建

Establishment of a murine model of alternaria alternate induced allergic asthma

目的研究使用互隔交链孢霉孢子构建C57BL/6小鼠肺部变态反应性炎症模型的方法。方法模型组、阳性对照组和阴性对照组各12只小鼠。模型组以互隔交链孢霉孢子悬液致敏和激发小鼠,分别于第0、5、10天腹腔注射霉菌孢子悬液,第14天开始连续4天鼻滴霉菌孢子悬液。阳性对照组以鸡卵蛋白(OVA)代替霉菌孢子,阴性对照组以磷酸缓冲液(PBS)代替霉菌孢子。进行肺组织病理检查,支气管肺泡灌洗液(BALF)中细胞计数分类和总蛋白浓度测定,ELISA测定BALF中的白细胞介素-4(IL-4)和干扰素-γ(IFN-γ)及血浆中的总IgE和特异性IgE,测定小鼠气道阻力。结果模型组和阳性对照组可见明显炎症细胞浸润,特别是嗜酸性粒细胞,而阴性对照组则未见。BALF中细胞总数和嗜酸性粒细胞比例较阴性对照组明显升高(P<0.05);BALF上清中IL-4、总蛋白水平较阴性对照组明显升高(P<0.05),IFN-γ水平较阴性对照组降低(P<0.05)。血浆中总IgE和特异性IgE较阴性对照组明显升高(P<0.05)。小鼠气道阻力较阴性对照相比有明显升高(P<0.05)。结论使用互隔交链孢霉孢子致敏和激发C57BL/6小鼠成功的建立了小鼠气道变态反应性炎症模型。

Objective To develop a method of establishment of airway allergic inflammation model of C57BL/6 mice sensitized and challenged with the spores of alternaria alternate. Methods C57BL/6 mice were divided into negative control group, positive control group and model group, twelve mice in each group. Mice of model group were sensitized with spores at day 0, 5, 10 by intraperitoneal injection, and challenged four times by intranasal drip from day 14 to day 17. OVA instead of spores were used in the positive control group, PBS used in the negative control group. Bronchoalveolar lavage fluid （BALF） was taken at 24h after the last time of challenge, and the pathological manifestation of the lung, cell counts and differential count and IL-4, IFN-y total protein levels in BALF were assessed. The levels of total IgE and specific IgE in plasma were identified, the airway resistance and lung compliance were determined. Results There was pulmonary eosinophilic inflammation in the positive control group and model group. Total cells count, differential cells count and IL-4 and total protein levels in BALF were enhanced significantly while IFN-y declined remarkably. The levels of total IgE and specific IgE were increased remarkably in plasma in positive control group and model group, the airway resistance was enhanced and lung compliance decreased. Conclusion An airway allergic inflammation model of C57BL/6 mice has been established by sensitizing and challenge with spores of alternaria alternate.