闭塞性细支气管炎小鼠模型的建立与评价

发布时间：2018-04-21 13:58

本文选题：闭塞性细支气管炎 + 丁二酮　；参考：《第三军医大学学报》2014年22期

【摘要】：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/m L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/m L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。
[Abstract]:Objective to establish an animal model of bronchiolitis obliterans Bos. Methods SPF grade C57BL/6 male mice were randomly divided into two groups: control group and model group with 6 rats in each group. The model group was constructed with the first intratracheal instillation of 490mol / L of 2-butanedione diketone, or 500 mg/kg)1, while the control group was replaced by the first intratracheal drip of disinfected distilled water. The other experimental conditions were the same as those of the two groups. The specimens were collected on the 3rd and 7th day after feeding in the SPF animal center. Bronchoalveolar lavage lavage were collected from bronchoalveolar lavage fluid (BALF) and the left lung was examined by pathology. During feeding, we observed the general state of the mice, monitored the changes of body mass in the two groups, monitored the airway hyperresponsiveness, bronchoalveolar lavage, cell count and inflammatory cell classification of the mice by noninvasive pulmonary function scintigraphy, and the left lung was stained with HE in pathological sections. Image analysis and collection. Results 1 day after DA administration, the mice in the model group developed wheezing and breathing quickening, and the body mass of the monitoring group was significantly lower than that of the control group (P 0.05). Compared with the control group, the pulmonary function (50 mg/m / L nebulized methacholine) in the model group was significantly increased by enhanced intermittent expiratory enhancement of Penhs (P 0.05). The total number and classification of BALF inflammatory cells in the model group were significantly higher than those in the control group on the 3rd day (P 0.05), the number of lymphocytes and macrophages on the 7th day was significantly higher than that in the control group (P 0.05). He staining of pathological sections showed that typical BO changed a large number of inflammatory cells in and around the bronchiole lumen of the model group at day 3.The lumen was severely occluded and the basal cell necrosis was severe in the model group after 7 days of acute hemorrhage. Although the airway epithelial cells attached to the basement, the cells were obviously hyperplastic and hypertrophic, with serious nuclear loss, inflammatory cell infiltration in the lumen, thickening of the lumen wall, and fibrosis. Conclusion the intratracheal instillation of DA(500 mg/kg)1 in C57BL/6 mice can observe the typical changes of BO in acute phase on the 7th day, and the model of BO mice can be established successfully.
【作者单位】：重庆医科大学附属儿童医院儿童发育疾病研究教育部重点实验室;重庆医科大学附属儿童医院重庆市干细胞治疗工程技术研究中心;重庆医科大学儿童医院临床分子医学中心;重庆医科大学附属儿童医院呼吸中心;
【基金】：重庆市卫生和计划生育委员会医学科研计划重点项目(20141008)~~
【分类号】：R-332;R562.21

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