闭塞性细支气管炎小鼠模型的建立与评价

闭塞性细支气管炎小鼠模型的建立与评价

李松1，邹琳2，符州3，王莉佳4，代继宏3

儿童发育疾病教育部重点实验室1，

重庆医科大学儿童医院临床分子中心2，

重庆医科大学儿童医院呼吸中心3

重庆医科大学儿童医院呼吸实验室4，

[摘要] 目的探讨建立闭塞性毛细支气管炎（BO）小鼠动物模型的方法。方法将40只 SPF级

C57BL/6雄性小鼠(6-8w)随机分为对照组和模型组两组。模型组的构建采用初次气管内滴注二乙酰（DA）（490mg/ml，500mg/kg）1次，对照组采用已消毒的蒸馏水代替，其余实验条件和方法均相同，

饲养于SPF级动物中心，至第3天和第7天收集标本。饲养期间观察小鼠一般状态，监测两组体重变化、肺功能改变；支气管肺泡灌洗、细胞计数及炎性细胞分类计数；病理切片、HE染色等。结果模

型组小鼠给予DA（500mg/kg）后1天可出现吼喘、呼吸加快等表现。模型组小鼠BALF液炎性细胞总

数及分类计数可见：3天以中性粒细胞为主，7天以淋巴和单核细胞为主（p＜0.05）；病理切片HE染

色可见典型的BO改变（气道上皮脱落严重；细支气管管腔内及管腔周围3天可见大量炎性细胞浸润，

伴肺泡急性出血，气道上皮大部分脱落，管腔严重闭塞；7天可见管壁增厚，炎症细胞浸润明显，类

纤维化表现）。结论 C57BL/6小鼠给予气管内滴注DA（500mg/kg）1次，可于第7天观察到典型的急

性期BO改变。

[关键词] 闭塞性细支气管炎；二乙酰；小鼠；模型

Establishment and evaluation of model of bronchiolitis obliterans

Li Song1, Zou Lin2, Fu Zhou3, Wang Lijia4, Dai Jihong3（1Key Laboratory of Developmental Diseases in Childhood of Ministry of Education, 2Center for Clinical Molecular Medicine, 3Center of Respiratory Disorders,

4Laboratory of Respiratory Disorders ,Children’s Hospital, Chongqing Medical University, Chongqing, 400014, China）

[Abstract] Objective Explore the establishment of bronchiolitis obliterans (BO) method

of mouse animal model. Methods Fourty C57BL/6 mice ( SPF )were randomly divided into normal control group and BO model group ,20 mice per group. BO group intratracheal 作者简介

李松，硕士研究生，E-mail：qwe130********@

通信作者代继宏，主任医师，博士，E-mail：danieljh@

instillation with Diacetyl(DA) (490mg/ml，500mg/kg),once.The control group with distilled water instead of sterile, the rest of the experimental conditions and methods are the same, keeping in SPF class animal center, to 3 and 7 days to collect specimens.Then General state

of the mice was observed during feeding, monitoring groups weight changes, lung function; bronchoalveolar lavage, cell counts and inflammatory cell count, pathology sections,HE staining. Results After the model group mice were given DA (500mg/kg) at the first day can occur roar breath, rapid breathing and other symptoms. BALF fluid and total number of inflammatory cells are observed that 3 days dominated by neutrophils, 7 days are main of lymphoid cells and monocytes (p<0.05); HE staining can see the typical BO changes (severe tracheal epithelium denudation; visible within the first three days lumen bronchial lumen and

the surrounding inflammatory cell infiltration, with acute alveolar hemorrhage, airway epithelial shedding most, severe occlusion of the lumen ;seventh days can visible wall thickening and inflammatory cells infiltration, similar fibrosis class performance). Conclusion Intratracheal instillation DA (500mg/kg) one time can establish a stable and typical murine model of bronchiolitis obliterans.

[Key words] bronchiolitis obliterans；diacetyl ; mouse; model

闭塞性细支气管炎(bronchiolitis obliterans,BO)是气道炎性损伤所致的慢性呼吸道疾病[1]，表现为小气道纤维增殖性病变使管腔狭窄，并最终闭塞，患者出现进行性、不可逆的气流受阻。BO是严重威胁人类健康的呼吸道疾病。对患者气道造成显著不可逆损害，目前尚无有效防治手段，所带来的社会卫生支出较高，应高度重视，积极寻找病因、发病机制及有效治疗手段。

成功的动物模型是研究BO发病机制、干预的重要手段，本研究将探讨建立BO小鼠模型的方法，观察病理学等指标改变，为今后的研究打下基础。

1 材料和方法：

1.1 实验材料：

选取6-8周清洁级雄性小鼠，C57BL/6，40只，购于重庆医科大学实验动物研究中心，按体重随机分为2组，动物饲养和实验均于动物实验室屏障环境进行，实验时室温(22±1℃) ，湿度(47±2）%。对照组:control组、模型组：BO组；小鼠饲养于重庆医科大学附属儿童医院实验动物研究中心，SPF动物房；