鼻内滴注烟草提取物和脂多糖建立慢性阻塞性肺疾病小鼠模型的评价

Evaluation on mouse models of chronic obstructive pulmonary disease induced by intranasal instillation of cigarette smoke extract and lipopolysaccharide

目的:建立鼻内滴注烟草提取物(CSE)和脂多糖(LPS)制备慢性阻塞性肺疾病(COPD)小鼠模型,并探讨其可行性。方法:24只C57BL/6J雄性小鼠随机分为对照组和模型组,每组12只,鼻内滴注CSE和LPS建立COPD小鼠模型,测定2组小鼠支气管肺泡灌洗液(BALF)中细胞总数、中性粒细胞数和巨噬细胞数,观察2组小鼠肺组织病理学表现并测定PAS阳性细胞数、肺泡平均内衬间隔(MLI)和肺泡破坏指数(DI),Western blotting法检测2组小鼠肺组织中E-cadherin、Vimentin和α-SMA蛋白表达水平。结果:实验4和6周时模型组小鼠BALF中细胞总数、中性粒细胞数和巨噬细胞数明显高于对照组(P<0.01),模型组小鼠肺组织出现典型粘液高分泌和肺气肿改变;实验4和6周时模型组小鼠PAS阳性细胞数、肺泡MLI(P<0.01)和DI(P<0.01)明显高于对照组;与对照组比较,实验4和6周时模型组小鼠肺组织中E-cadherin蛋白表达水平降低(P<0.01),Vimentin和α-SMA蛋白表达水平升高(P<0.01)。结论:采用鼻内滴注CSE和LPS的方法可在较短时间内建立COPD小鼠模型。

Objective:To constract the mouse models of chronic obstructivePulmonary disease(COPD)induced by intranasal instillation of cigarette smoke extract(CSE)and lipopolysaccharide(LPS),and to explore its feasibility.Methods:Twenty-four C57BL/6J male mice were randomly divided into control group and model group,and there were 12 mice in each group.The COPD mouse model was established by intranasal instillation of CSE and LPS.The number of total cells,neutrophils and macrophages in bronchoalveolar lavage fluid(BALF)of the mice in two groups were determined,thePathological changes of lung tissue of the mice in two groups were observed,and the number ofPASPositive cells,alveolar mean lining interval(MLI)and alveolar damage index(DI)(P<0.01)of the mice in two groups were determined.Western blotting method was used to detect the expression levels of E-cadherin,Vimentin,andα-SMAProteins in lung tissue of the mice in two groups.Results:The number of total cells,neutrophilis and macrophages in BALF of the mice in model group at 4 and 6 weeks of experiment was significantly higher than those in control group(P<0.01),and the typical mucus hypersecretion and emphysema in lung tissue of the mice in model group were found;the number ofPASPositive cells,MLI(P<0.01),and DI(P<0.01)of the mice in model group at 4 and 6 weeks were higher than those in control group.Compared with control group,the expression level of E-cadherinProtein in lung tissue of the mice in model group was decreased(P<0.01),and the expression levels of Vimentin andα-SMAProteins in lung tissue of the mice in model group were increased(P<0.01).Conclusion:The mouse model of COPD is successfully established by intranasal instillation of CSE and LPS in a shortPeriod of time.