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纸质出版:2014
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李松,张素倩,邹琳,符州,代继宏.闭塞性细支气管炎小鼠模型的建立与评价[J].陆军军医大学学报(原第三军医大学学报),2014,36(22):2283-2286.
Li Song,Zhang Suqian,Zou Lin,Fu Zhou,Dai Jihong.Establishment and evaluation of murine model of bronchiolitis obliterans[J].J Amry Med Univ (J Third Mil Med Univ),2014,36(22):2283-2286.
目的 探讨建立闭塞性细支气管炎(bronchiolitis obliterans
BO)小鼠动物模型的方法。 方法 将SPF级C57BL/6雄性小鼠(6~8周)按随机数字表法分为两组:对照组和模型组(BO组),每组6只。模型组构建采用初次气管内滴注2
3丁二酮(DA)(490 mg/mL即500 mg/kg)1次,对照组采用初次气管内滴注已消毒的蒸馏水代替,其余实验条件和方法2组相同。饲养于SPF级动物中心,至第3天和第7天收集标本
分别留取支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)和左肺行病理学检测。饲养期间观察小鼠一般状态,监测两组体质量变化,用无创肺功能体描仪行小鼠气道高反应监测,支气管肺泡灌洗,细胞计数及炎性细胞分类计数;左肺行病理切片、HE染色、图像分析与采集。 结果 模型组小鼠给予DA后1 d可出现吼喘、呼吸加快等,监测体质量显著低于对照组(P0.05)。与对照组相比,模型组肺功能(雾化乙酰甲胆碱浓度为50 mg/mL时)增强呼气间歇(enhanced pause
Penh)值明显升高(P0.05)。模型组小鼠BALF炎性细胞总数及分类计数可见第3天时中性粒细胞总数显著高于对照组(P0.05),第7天时淋巴细胞总数、巨噬细胞总数显著高于对照组(P0.05)。病理切片HE染色可见典型的BO改变。3 d模型组细支气管腔内及管腔周围大量炎性细胞浸润,管腔严重闭塞,组织急性期出血严重;7 d模型组基底细胞严重坏死,气道上皮细胞虽附着于基底,但可见细胞明显增生肥大,细胞核丢失严重,管腔内炎性细胞浸润,管腔壁增厚,类纤维化表现。 结论 C57BL/6小鼠给予气管内滴注DA(500 mg/kg)1次,可于第7天观察到典型的急性期BO改变,可以成功建立BO小鼠模型。
Objective To explore the establishment of murine model method for bronchiolitis obliterans (BO). Methods C57BL/6 mice(SPF) were randomly divided into normal control group and BO model group and 6 mice per group. The mice of BO group were intratracheal instillated with diacetyl (DA) (490 mg/mL
500 mg/kg) and the mice in control group with sterile distilled water. The rest of the experimental conditions and methods were the same. The mice were kept in SPF class animal center for 3 or 7 d to collect specimens. The mice were dynamically observed the general status
body weight
pulmonary function
cell counts
inflammatory cell count of bronchoalveolar lavage fluid
as well as the pathological and HE staining for lung slices. Results The model group mice were given DA (500 mg/kg) after 1 d and they presented wheezing
rapid breathing and other symptoms
the weight was significantly lower than the control group (P0.05). Compared with the control group
the lung function (aerosolized methacholine concentration was 50 mg/mL) enhanced pause (Penh) was increased significantly (P0.05). Model mice inflammatory cell count and classification counting neutrophils were significantly higher than the control group (P0.05) in 3 d
and the total cells and lymphocytes
macrophages were significantly higher than that of the control group (P0.05) in 7 d. Pathological section HE staining showed typical BO changes (around the 3 d, model group bronchial lumen and infiltration of inflammatory cells
luminal severe occlusion
tissue in the acute stage of severe bleeding; In 7 d
basal cells were severe necrosis
airway epithelial cells were attached to the substrate
but seen obvious cell hyperplasia and hypertrophy
cell nucleus lost seriously
inflammatory cell infiltration in the lumen
and the lumen wall thickening
fibrosis). Conclusion The mouse model of bronchiolitis obliterans (BO) should be established successfully by intratracheal instillation DA (500 mg/kg) one time.
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