纳米二氧化硅与微米二氧化硅致大鼠肺纤维化miRNAs差异表达比较研究  

Comparison study on the differential expression of miRNAs in rat pulmonary fibrosis induced by nanosized SiO2 and microsized SiO2

作　　者：杨红 李明月 李文超 张迎建 劳灿山 Yang hong;Li Mingyue;Li Wenchao;Zhang Yingjian;Lao Canshan(Key Laboratory of Environmental Medicine Engineering of Ministry of Education,School of Public Health,Southeast University,Nanjing 210009,China)

机构地区：[1]东南大学公共卫生学院环境医学工程教育部重点实验室,南京210009

出　　处：《中华劳动卫生职业病杂志》2019年第2期81-89,共9页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金面上项目(81273046);江苏省预防医学科研课题(Y2012039);中央高校基本科研业务费专项基金.

摘　　要：目的比较纳米二氧化硅(SiO2)与微米二氧化硅(SiO2)致大鼠肺纤维化过程中miRNAs差异表达的变化,分析miRNAs表达水平与肺纤维化的相关性。方法将36只体重为180~220 g的SPF级健康雄性SD大鼠随机分为对照组、纳米SiO2染尘组和微米SiO2染尘组,每组12只,用一次性气管灌注染毒方法分别经气管灌注1ml生理盐水、25mg/ml纳米SiO2溶液和25mg/ml微米SiO2溶液。分别于染尘60、90 d各处死6只大鼠,用光学显微镜和透射电镜观察大鼠肺组织病理学及超微结构改变;用IlluminaHiSeq 2000高通量测序技术分析大鼠肺组织miRNA表达谱;用Targetscan对miRNAs进行靶基因预测,通过基因本体论Gene Ontology(GO)和京都基因与基因组百科全书(KEGG)分别对靶基因进行功能分类和pathway分析,筛选与肺纤维化相关的靶基因。结果光学显微镜下可见,染尘60、90 d时,纳米SiO2染尘组大鼠肺组织支气管和血管周围、肺泡壁和小叶间隔增宽,纤维结缔组织轻度增生。染尘60 d时,微米SiO2染尘组大鼠肺内弥漫性分布细胞性结节,多数结节融合,纤维母细胞增生明显,结节周围肺泡代偿性肺气肿;染尘90 d时,微米SiO2染尘组大鼠肺内弥漫性分布的细胞纤维性结节内纤维成分增加,融合的大结节周围肺泡代偿性肺气肿。透射电镜下可见,染尘60、90 d时,纳米SiO2组大鼠肺组织Ⅱ型肺泡上皮细胞嗜锇板层小体增多、肿胀,空泡样变,间质中胶原纤维及弹力纤维增生;微米SiO2染尘组大鼠肺组织Ⅱ型肺泡上皮细胞胞质内嗜锇板层小体增多,间质中胶原纤维及弹力纤维增生,嗜锇板层小体空泡样变。与对照组比较,染尘60、90d时,纳米SiO2染尘组大鼠肺组织中差异上调表达miRNA分别为50、72个,下调表达miRNA为22、24个;微米SiO2染尘组差异上调表达miRNA分别为91、70个,下调表达miRNA为34、78个。两者共同差异上调表达的miRNA为miRNA-18a、miRNA-702-3p,下调表达的miRNA为miRObjective To explore the correlation between expression level of miRNAs and pulmonary fibrosis on the basis of comparison the differential expression of miRNAs in rat pulmonary fibrosis induced by nano SiO2 and micron SiO2.Methods Thirty-six healthy male SD rats weighting 180-220 g were randomly divided into 3 groups.They were instilled intratracheally with 1 ml suspension of saline,25 mg/ml nanosized SiO2 and microsized SiO2 particles and sacrificed at 60 d and 90 d postexposure from each group with six rats.The change of pathological morphology and ultrastructure of lung were observed by optical and transmission electron microscopy.The differentially expressed microRNAs in lung tissue of the rats after instilled intrachcally nanosized SiO2 and microsized SiO2 particles at 60 d and 90 d were determined by Illumina HiSeq 2000 sequencing technique.Target prediction for miRNAs was conducted by databases of Target-scan.Function-significant enrichment analysis and signal pathway analysis for predicted target genes were respectively conducted by the GO and the KEGG,then target genes related to pulmonary fibrosis were screened out.Results Light microscope examination showed that wide bronchi,vessels,interlobular septa and slight fibrous connective tissue proliferation at 60 d and 90 d postexposure in 25 mg/ml nanosized SiO2 group.A few fused nodules at 30 d postexposure,a lot of fused nodules at 60 d postexposure,fibrous cell nodules and compensatory emphysema around alveolar at 90 d postexposure in 25 mg/mL microsized SiO2 group were observed.Electron microscopy demonstrated swelling and vacuolar degeneration of osmiophilic lamellar bodies in typeⅡalveolar epithelial cells,collagen fiber and elastic fiber hyperplasia in pulmonary interstitial at 60 d,90 d postexposure in 25 mg/ml nanosized SiO2 group.Increased and vacuoloid changed osmiophilic lamellar bodies in typeⅡalveolar epithelial cells,collagen fiber and elastic fiber hyperplasia in the interstitial at 60 d,90 d postexposure in 25 mg/ml microsized SiO2 grou

关 键 词：纳米二氧化硅 微米二氧化硅 肺纤维化 MICRORNA表达谱 

分 类 号：R563[医药卫生—呼吸系统]