过敏性肺炎动物模型制作

Allergic Pneumonia Animal Model Manufacture

目的 探讨毛松茸孢子抗原对小黑鼠是否能引起过敏性肺炎及制作过敏性肺炎动物模型的方法。方法 选取6～8周龄雄性、体质量25～30gC57BL／6小黑鼠60只。在鼠尾注射松茸孢子抗原和免疫佐剂（Freud’s adjuvant）复合物，进行1次全身抗原攻击，通过气管注入一定量抗原（1次／周）以进行局部抗原攻击。60只小黑鼠分为A、B、C、D组，每组分别为10、10、20、20只。A组为正常对照组；B组予Freud’s adjuvant，方法同上。C、D组分别向呼吸道注入孢子抗原2和4次。当抗原攻击结束1周后C、D组每组又分2组，每组各10只。其中1组吸入1．5％孢子抗原，以诱发急性反应后收集肺泡灌洗液（BALF），观察各种细胞百分率，摘除肺组织制作病理标本；另一组未经抗原诱发急性反应而直接收集BALF和摘除肺组织；B组吸入生理盐水后收集BALF和摘除肺组织作对照。酶联免疫吸附法（ELISA）测定小黑鼠血清抗原特异性IgG。结果 C和D组血清抗原特异性IgG较A和B组显著增高（P均〈0．05）。C和D组BALF中出现淋巴细胞和中性粒细胞，D组还出现少量嗜酸性粒细胞，且两组淋巴细胞和中性粒细胞较A和B组显著增高（P均〈0．05）。C和D组肺组织切片均出现淋巴细胞和中性粒细胞浸润，D组出现肺纤维化倾向。结论 孢子抗原导致小鼠血清特异性IgG增高，肺组织大量白细胞浸润及纤维化。即毛松茸孢子抗原可在C57BL/6小黑鼠引起典型过敏性肺炎。

Objective To investigate whether the spores from mushroom antigen can cause the allergic pneumonia and manufacture allergic animal model in the C57BL/6 mouse. Methods Aged 6 weeks old,weight 25 - 30 g C57BL/6 mice were collected. In the mouse tail injection compound of spore antigen and the Freud＇s adjuvant. Then pours into through the trachea the antigen once a week. The mice were divided into 4 groups. Group A was the normal mouse,Group B was given Freud＇s adjuvant （the same method）to determine whether there was affect to the mouse. Group C and D were injected spore antigen 2 and 4 times. When the antigen sensitization finished 1 week later group C and D were completely divided 2 groups,among them one group was inhalation 1.5% spore antigen and induce the acute response. Six hours later the bronchoalveolar lavage fluid（BALF） was collected to observe cell change,and excise the lung tissue to manufacture the pathology specimen,another group had not been induced the acute response and collect the BALF and to exsise the lung tissue directly. Group B were inhalted saline later to collect the BALF and the lung tissue. In the mouse blood serun,enzyme linked immunosorbent assay（ELISA） was used to mensurate antigen specific IgG. Results In group C and D, antigen specific IgG significantly inhanced than that in group A and B （all P 〈 0.05）. In BALF of group C and D, the lymphocyte and the neutrophil were observed, group D also appeared few eosinophil;also 2 groups of lymphocyte and neutrophil remarkably increased than group A and B （all P〈0.05）. In the lung tissue slice, group C and D all appeared the lymphocyte and neutrophil infiltrating,group D appeared the lung fibrosied tendency. Conclusions The mushroom spore antigen has increased serum differential IgG, many leucocyte infiltration and fibrosis in the lung tissue, so it has caused the pathologic change of allergic pneumonia in the C57BL/6 mouse.