熏香烟加气管注入脂多糖和结合臭氧暴露法建立大鼠慢性阻塞性肺病模型

Fumigation and intratracheal instillation of lipopolysaccharide or they combining ozone exposure for establishing COPD models in rats

目的评价用熏香烟加气管注入脂多糖(LPS)和在此基础上结合臭氧暴露的方法建立的大鼠慢性阻塞性肺病(COPD)模型。方法随机数表法将30只SD雄性大鼠分为正常对照组、模型Ⅰ组和模型Ⅱ组,采用熏香烟加气管注入LPS法和在此法基础上结合臭氧暴露的方法分别复制大鼠COPD模型。观察大鼠的一般情况、COPD发作的症状,测定3组的肺功能。苏木素-伊红(HE)染色观察大鼠肺组织形态变化和炎症细胞浸润情况等,并分析平均内衬间隔(MLI)和平均肺泡数(MAN)。结果两种方法所得模型均较理想,表现为肺功能检测显示与空白对照组比,模型Ⅰ、Ⅱ组中心气道阻力(Rn)及组织阻尼(G)升高,有统计学差异(P<0.05),提示2组均有气流受限及换气功能障碍。肺组织HE染色镜下可见各级支气管黏膜上皮肿胀、增生及脱落损伤,气道与肺实质大量炎细胞浸润,气道壁、肺小动脉壁增厚,肺大泡形成等多种改变,比较模型Ⅰ、Ⅱ组,模型Ⅱ组病理损伤更重,表现为支气管黏膜上皮脱落更彻底,炎细胞浸润更广泛,气道壁、肺小动脉壁增厚更明显及肺泡融合破坏更严重等。MLI及MAN结果为单位面积内模型Ⅰ组和Ⅱ组MAN低于空白对照组,而MLI大于空白对照组,差异有统计学意义(P<0.01)。结论用熏香烟加气管注入LPS和结合臭氧暴露法均可重现COPD组织病理特征,可成功复制较理想的大鼠COPD模型,叠加臭氧暴露之后肺部病理损伤更为严重,可能更能模拟临床实际情况。

Objective To review rat COPD models established by using fumigation ＋ intratracheal instillation of lipopolysaccharide （LPS） or fumigation ＋ LPS combining ozone exposure. Methods Male SD rats （n = 30 ） were randomly divided into blank group, model Ⅰ group and model Ⅱ group. The general condition and symptoms of COPD were observed in rats. The morphological changes of lung tissue and inflammatory cell infiltration were observed after HE staining, and results of mean lung interval （MLI） and mean alveoli number （MAN） were observed and analyzed after HE staining. Results COPD models established by using two methods was satisfactory. The detection of lung function showed that central airway resistance （Rn） and organization damping （G） increased in model Ⅰ group and model Ⅱ group compared with blank group （P 〈 0. 05 ）, which indicated there were airway limitation and ventilation dysfunction in model Ⅰ group and model Ⅱ group. The swelling, hyperplasia and exfoliation of all levels of bronchial mucosal epithelium, serious inflammatory cell infiltration in airway and lung tissue, thickened of walls of airway and pulmonary arterioles and bullae of lung were observed after HE staining. All of these pathological changes were more serious in model Ⅱ group compared with model Ⅰ group. The results of MLI and NAN showed that MAN was lower and MLI was higher within unit area in model Ⅰ group and model Ⅱ group than that in blank group P 〈 0.05 ）. Conclusion Fumigation and intratracheal instillation of lipopolysaccharide or they combining ozone exposure can reappear the pathological features of COPD and establish successfully rat COPD models. After combining ozone exposure, the pathological damages in lung are more serious, which is more approximate to clinical situation.