一种改良的博来霉素诱导的肺纤维化模型造模方法  

An improved modeling method of bleomycin-induced pulmonary fibrosis

作　　者：李乔 王苗风 田宇 赵峰 LI Qiao;WANG Miaofeng;TIAN Yu;ZHAO Feng(Department of Respiratory and Critical Care Medicine,Xijing Hospital,Air Force Medical University,Xi'an 710032,China)

机构地区：[1]空军军医大学西京医院呼吸内科,陕西西安710032

出　　处：《空军军医大学学报》2024年第4期425-429,共5页Journal of Air Force Medical University

基　　金：国家自然科学基金面上项目(81570054)。

摘　　要：目的建立博来霉素诱导的小鼠肺纤维化模型,比较经气管滴注和经气管吸入两种不同给药方式的成模差异。方法选择6~8周C57BL/6雄性小鼠46只,取6只小鼠随机分为经气管滴注组和经气管吸入组,每组3只。小鼠麻醉后分别经气管滴注和吸入亚甲蓝,观察小鼠双肺和胃内亚甲蓝的分布。取另40只小鼠随机分为经气管滴注+生理盐水组、经气管吸入+生理盐水组、经气管滴注+博来霉素组、经气管吸入+博来霉素组。观察各组小鼠造模后的精神状态、体质量变化和生存状况,并在第21日取肺组织用于组织学、羟脯氨酸和TGF-β、α-SMA和COL1A1等蛋白水平Western blotting检测。结果经气管滴注组,亚甲蓝不均匀且局限地分布于双肺,伴不同程度胃内残留。经气管吸入组,亚甲蓝较均匀且弥散地分布于双肺,无明显胃内残留。与两对照组相比,两模型组小鼠精神状态欠佳,体质量下降明显,死亡率升高;小鼠肺组织肺泡结构破坏严重,伴大量胶原沉积,羟脯氨酸含量显著升高(P<0.05),Western blotting显示肺组织TGF-β、α-SMA和COL1A1等蛋白的表达显著增多(P<0.05)。结论经气管吸入+博来霉素组小鼠纤维化病灶分布更加均匀,各项指标相对稳定,是建立肺纤维化模型理想的方法。Objective To establish bleomycin-induced pulmonary fibrosis model in mice,and compare the differences of two different administration modes:transtracheal instillation and transtracheal inhalation.Methods A total of 46 male C57BL/6 mice aged 68 weeks were selected,and 6 mice were randomly divided into transtracheal instillation group and transtracheal inhalation group,with 3 mice in each group.The distribution of methylene blue in the lungs and stomach of the mice was observed after they were instilled and inhaled methylene blue after anesthesia.Another 40 mice were randomly divided into transtracheal instillation+normal saline group,transtracheal inhalation+normal saline group,transtracheal instillation+bleomycin group,and transtracheal inhalation+bleomycin group.The mental state,body mass changes and survival status of mice in each group were observed after modeling,and lung tissues were collected on the 21st day for histology,hydroxyproline and TGF-β,α-SMA and COL1A1 protein levels by Western blotting.Results In the transtracheal instillation group,methylene blue was distributed unevenly and restrictively in both lungs,with varying degrees of gastric residue.In the transtracheal inhalation group,methylene blue was distributed evenly and diffusively in both lungs,without obvious gastric residue.Compared with the two control groups,the two model groups had poor mental state,significant body mass loss,high mortality,severely damaged alveolar structure of lung tissue of mice,a large amount of collagen deposition,and significantly increased content of hydroxyproline(P<0.05).Western blotting showed that the expressions of TGF-β,α-SMA,and COL1A1 proteins in lung tissue were significantly increased(P<0.05).Conclusion The fibrotic lesions of transtracheal inhalation+bleomycin group are more evenly distributed and the indexes are relatively stable,which is an ideal method to establish pulmonary fibrosis model.

关 键 词：肺纤维化 博来霉素 经气管给药 疾病模型 

分 类 号：R563[医药卫生—呼吸系统] R-332[医药卫生—内科学]