摘要
目的探讨肺部菌群失调对矽肺小鼠肺纤维化和肺上皮间质转化(EMT)过程的影响。方法将无特定病原体级雄性C57BL/6小鼠随机分为空白对照组、矽肺模型组、溶剂对照组、万古霉素(VM)+氨苄青霉素(AMP)组、甲硝唑(MNZ)+新霉素(NEO)组和混合处理组,每组12只。除空白对照组予20.0μL的0.9%氯化钠溶液外,其余5组小鼠均采用一次性气管滴注法滴注20.0μL质量浓度为250.0 g/L的矽尘混悬液建立矽肺小鼠模型。各组小鼠采用鼻内滴注法对应予以下处理:溶剂对照组小鼠予双蒸水;VM+AMP组小鼠予质量浓度为0.5 g/L的VM和1.0 g/L的AMP;MNZ+NEO组小鼠予质量浓度均为1.0 g/L的MNZ和NEO;混合处理组小鼠予上述相同剂量的四种抗生素,滴注体积均为50.0μL;于染尘前7 d、染尘前0.5 h以及染尘后7、14、21 d各滴注抗生素1次。染尘后28 d采集小鼠肺组织,采用苏木精-伊红和Masson染色法观察肺组织病理变化;以蛋白质印迹法检测肺组织中α平滑肌肌动蛋白(α-SMA)、E-钙黏蛋白(E-CAD)、波形蛋白(VIM)的相对表达水平,免疫组织化学法检测肺组织中E-CAD和VIM相对表达水平,实时荧光定量聚合酶链反应法检测肺组织中Ⅰ型胶原α2链(Col1a2)mRNA相对表达水平。结果组织病理学结果显示,空白对照组小鼠肺泡壁薄且结构完整,周围浸润炎性细胞少,无异常的胶原纤维分布;矽肺模型组小鼠肺泡结构紊乱,伴有大量炎性细胞浸润,肺泡壁增厚,细胞纤维结节增生,蓝色胶原沉积大量分布在纤维结节团块中;与矽肺模型组和溶剂对照组比较,VM+AMP组、MNZ+NEO组和混合处理组小鼠肺组织炎症与纤维化均有不同程度的减轻。与空白对照组比较,矽肺模型组小鼠肺组织α-SMA、VIM蛋白和Col1a2 mRNA相对表达水平均升高(P值均<0.05),E-CAD蛋白相对表达水平降低(P<0.05);与矽肺模型组和溶剂对照组比较,MNZ+NEO组和混合处理组小鼠肺组织α-SMA、VIM蛋白和Col1a
Objective To investigate the effect of lung flora dysbiosis on the process of pulmonary fibrosis and lung epithelial-mesenchymal transition(EMT)in mice with silicosis.Methods Male C57BL/6 mice of specific pathogen-free grade were randomly divided into the blank control group,silicosis model group,solvent control group,vancomycin(VM)+ampicillin(AMP)group,metronidazole(MNZ)+neomycin(NEO)group and mixed treatment group,12 mice in each group.Except for the blank control group,which was given 20.0μL of 0.9%NaCl solution,the other five groups of mice were dosed with 20.0μL of silica dust suspension at a mass concentration of 250.0 g/L using a single tracheal drip to establish the silicosis mouse model.The intranasal drip method was used to treat silicosis mice in each group as following:mice in the solvent control group were given double-distilled water;mice in the VM+AMP group were given VM at a mass concentration of 0.5 g/L and AMP at 1.0 g/L;mice in the MNZ+NEO group were given MNZ at a mass concentration of 1.0 g/L and NEO at 1.0 g/L;mice in the mixed treatment group were given the same doses of the four antibiotics mentioned above,all in a drip volume of 50.0μL.Silicosis mice were treated seven days and half an hour before silica dusting and 7,14,and 21 days after silica dusting.Mouse lung tissue was collected aseptically 28 days after silica dusting.Hematoxylin-eosin and Masson trichrome staining methods were used to observe the pathological changes.Western blotting was used to detect the relative protein expression ofα-smooth muscle actin(α-SMA),E-cadherin(E-CAD)and vimentin(VIM).Immunohistochemistry was used to detect the relative expression of E-CAD and VIM.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression levels of collagen typeⅠalpha 2(Col1a2)mRNA in lung tissues.Results The histopathological results showed that the alveoli of the blank control group were thin and structurally intact,with few surrounding infiltrating inflammatory cells and no abnormal dist
作者
彭金碧
韩宇浩
顾倚岑
荆茹
杨道宇
全宁斌
张家祥
王雨浩
李旭东
PENG Jin-bi;HAN Yu-hao;GU Yi-cen;JING Ru;YANG Dao-yu;QUAN Ning-bin;ZHANG Jia-xiang;WANG Yu-hao;LI Xu-dong(Department of Occupational and Environmental Health,School of Public Health,Anhui Medical University,Hefei,Anhui 230032,China;不详)
出处
《中国职业医学》
CAS
北大核心
2022年第5期514-521,共8页
China Occupational Medicine
基金
国家重点研发计划项目(2022YFC2503205)
西部绿色建筑国家重点实验室项目(LSKF202313)
广东省自然科学基金(2023A1515011329)
广东省职业健康工程技术研究中心(D:2019A069)
广东省职业病防治重点实验室(2017B030314152)
广州市基础研究计划项目(202002030510030008)
关键词
矽肺
肺纤维化
上皮间质转化
肺部菌群
失调
肺共生菌
小鼠
Silicosis
Pulmonary fibrosis
Epithelial-mesenchymal transition
Pulmonary flora
Dysbiosis
Lung commensal bacteria
Mice
