摘要
目的 比较硫磺烟熏结合冷水浴、香烟烟熏及脂多糖(LPS)雾化3种方法诱导小鼠肺炎的组织学改变及病理特征.方法 雄性昆明小鼠40只随机均分为对照组(正常喂养,不做其他处理)、SO2组(硫磺烟熏结合冷水浴,烟熏2.4 g/d)、香烟烟熏(XY)组(烟熏30根/d)及LPS组(LPS雾化10 mg/d),持续15 d;造模结束后,观察各组小鼠精神状态等一般情况及呼吸系统症状,麻醉处死小鼠取肺组织称肺湿重以计算肺脏系数,同时制作切片采用苏木精-伊红(HE)染色、马松(Masson)染色及阿利新蓝-过碘酸雪夫氏(Ab-pas)染色检测肺组织形态学特征,采用免疫组织化学染色法检测肺组织白细胞介素-1β(IL-1β)蛋白、核因子-κBp65(NF-κB p65)蛋白及α-平滑肌肌动蛋白(α-SMA)、胶原蛋白Ⅰ(Collagen-Ⅰ)分别评估炎症病理改变及胶原增殖、纤维化情况.结果 与对照组相比,其余各组小鼠均出现不同程度活动度减少、饮食减少、体质量下降、毛发枯黄无光泽且伴有咳嗽、打喷嚏和痰鸣音等呼吸系统症状;与对照组比较,SO2 组和LPS组小鼠肺脏系数升高(P＜0.05);HE、Masson和Ab-pas染色结果显示,SO2组和XY组小鼠肺组织中部分肺泡腔内和支气管周围见炎症细胞浸润、肺间质轻度纤维化增生,LPS组小鼠肺泡腔内见大量炎症细胞浸润、支气管周围及肺间质酸性黏蛋白分泌、肺间质明显纤维化增生;免疫组织化学染色结果显示,与对照组相比,SO2组、XY组及LPS组小鼠肺组织IL-1β蛋白表达均增加(P＜0.05),LPS组小鼠肺组织NF-κB p65、α-SMA及Collagen-Ⅰ蛋白表达增加(P＜0.05).结论 与其它方法相比,脂多糖雾化诱导小鼠肺组织病理形态学改变最接近人类肺炎的疾病特点,是较为理想的肺炎小鼠造模方法.
Abstract
Objective To compare thehistologic change and pathological features of three modeling methods[inducing sulfur smoking combined with cold water bath,cigarette smoking and lipopolysaccharide(LPS)atomization]to induce micepneumonia and to optimize the ideal modeling method.Methods Forty male Kunming mice were randomly divided into the control group(withnormal feedingonly),SO2 group(sulfur smoking combined with cold water bath,2.4 g/d),cigarette smoking(XY)group(30 pieces/d),and LPS group(LPS atomization 10 mg/d).The treatment for the 3 groups lasted for 15 days.After the modeling,the mental state,diet,body mass,hair luster and respiratory disease symptoms were observed.All the mice in each group were put to death under anesthesia.Lung tissues were taken and weighed by lung wet weight to calculate lung coefficient.Lungsections were prepared and the morphological characteristics of lung tissues were detected by hematoxylin-Eosin(HE)staining,Masson staining and Alcian blue-periodate Scheuff's(Ab-pas)staining.The pathological changes of inflammation,Collagen proliferation and fibrosis were evaluated by immunohistochemical staining of interleukin-1β(IL-1β)protein,nuclear factor-κB p65(NF-κB p65)protein,α-smooth muscle actin(α-SMA),and Collagen-Ⅰ.Results Compared with the control group,the other groups showed different degrees of decrease inactivity,dietand body mass,andwith withered hair.The respiratory symptoms such as cough,sneezing and sputum chirping.Compared with the control group,the lung coefficient of mice in SO2 group and LPS group increased(P＜0.05).HE,Masson and Ab-pas staining showed that in SO2 and XY groups,inflammatory cell infiltration and mild fibrosis and hyperplasia of interstitial lung were observed in some alveolar cavities and around bronchus,while in LPS group,a large number of inflammatory cell infiltration,acidic mucin secretion and obvious fibrosis and hyperplasia of interstitial lung were observed in alveolar cavities.Theimmunohistochemical staining showed that,compared with the control group,the expression of IL-1β protein in lung tissues of mice in SO2,XY,and LPS groups increased(P＜0.05),and the expression of NF-κB p65,α-SMA,and Collagen-Ⅰ protein in lung tissues of mice in LPS groupincreased(P＜0.05).Conclusion Compared with other two methods,the pathological changes of lung tissues induced byLPSatomization are mostlysimilar to the characteristics of human pneumonia,and it is an ideal method for pneumonia modeling in mice.
基金项目
贵州省科技计划(黔科合支撑[2020]4Y209)
维普
万方数据