miRNA-29b-3p和miRNA-34c-3p在染矽尘大鼠肺组织和A549细胞中表达变化的研究  

Expression changes of miRNA-29b-3p and miRNA-34c-3p in lung tissue of rats exposed to silica and A549 cells

作　　者：王佳宇 耿晓[2] 贾强[2] 李超 赛林霖[2] 于功昌[2] 邵华[2] Wang Jiayu;Geng Xiao;Jia Qiang;Li Chao;Sai Linlin;Yu Gongchang;Shao Hua(School of Medicine and Life Sciences,University of Jinan-Shandong Academy of Medical Sciences,Jinan 250062,China;Shandong academy of medical science shand and Occopational Medical,Jinan 250062,China)

机构地区：[1]济南大学山东省医学科学院医学与生命科学学院,250062 [2]山东省医学科学院山东省职业卫生与职业病防治研究院,济南250062

出　　处：《中华劳动卫生职业病杂志》2019年第2期110-115,共6页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：山东省重点研发计划项目(2016GSF201047、2017GSF18186);山东省医学科学院医药卫生科技创新工程;山东省自然科学基金项目(ZR2015YL046).

摘　　要：目的通过检测染矽尘大鼠肺组织和A549细胞中微RNA-29b-3p(miRNA-29b-3p)和微RNA-34c-3p(miRNA-34c-3p)的表达水平,探讨miRNA-29b-3p和miRNA-34c-3p在肺纤维化进程中的作用。方法SPF级雄性Wistar大鼠60只,按体重随机分为对照组和二氧化硅(SiO2)染尘组,每组30只,用一次性非暴露式气管滴注法灌注SiO2染尘组大鼠1 ml SiO2悬浊液,对照组大鼠相同方法气管内注入1 ml灭菌生理盐水。于染尘后1、7、14、21、28 d两组分别采集6只大鼠肺组织,其中3只大鼠取出肺组织进行病理学观察,另外3只采用miRNA微阵列芯片技术筛选肺组织中差异表达的miRNA。在体外细胞水平中培养A549细胞,分为对照组、SiO2刺激组和TGF-β1刺激组,在处理后的12、24和48 h收取细胞,通过荧光定量PCR(qRT-PCR)方法验证miRNA-29b-3p和miRNA-34c-3p在各组大鼠肺组织和A549细胞中的表达水平,对miRNA-29b-3p和miRNA-34c-3p进行靶基因预测并进行GO富集分析和KEGG通路分析。结果对照组大鼠体重增长速度较SiO2染尘组大鼠明显;与对照组比较,SiO2染尘组大鼠的肺脏质量和肺系数明显升高,差异有统计学意义(P<0.05)。对照组大鼠肺组织21、28 d炎症反应较1、7、14 d明显减轻,SiO2染尘组大鼠肺组织有持续性炎性细胞浸润;对照组大鼠肺组织21、28 d有少量胶原分布,SiO2染尘组大鼠肺组织21 d后开始出现大量胶原纤维沉积。与对照组比较,SiO2染尘组中miRNA-29b-3p和miRNA-34c-3p表达水平均明显下调,差异均有统计学意义(P<0.05)。A549细胞在SiO2和人重组TGF-β1处理后,与对照组比较,24 h和48 h处理组中miRNA-29b-3p和miRNA-34c-3p表达水平均明显下调,差异有统计学意义(P<0.05)。结论大鼠肺组织和A549细胞中miRNA-29b-3p和miRNA-34c-3p的下调可能与早期矽肺的发生发展有关,有望成为早期矽肺诊断以及预后的指标。Objective To investigate the role of microRNA-29b-3p(miRNA-29b-3p)and miRNA-34c-3p in the process of pulmonary fibrosis,we detected the expression levels of miRNA-29b-3p and miRNA-34c-3p in the lung tissue of rats exposed to silica and A549 cells.Methods SPF male Wistar rats were randomly divided into 1,7,14,21,28 d control group and silica(SiO2)dusting group,with 6 rats in each group.One-time non-exposure method was used to infuse 1ml SiO2 suspension.The rat SiO2 dusting group was established in the liquid,and the control rats were intratracheally injected with 1 ml of sterile physiological saline in the same manner.The lung tissues of each group were collected at the corresponding time points after dusting.Three of the rats were taken out for pathological observation,and the other three were used to screen differentially expressed miRNAs in lung tissue by miRNA microarray technology.A549 cells were cultured at the in vitro cell level and divided into control group,SiO2 stimulation group and TGF-β1 stimulation group,and cells were collected at 12,24 and 48 h after treatment.The expression levels of miRNA-29b-3p and miRNA-34c-3p in rat lung tissue and A549 cells were verified by real-time PCR(qRT-PCR),target gene prediction of miRNA-29b-3p and miRNA-34c-3p and perform GO enrichment analysis and KEGG pathway analysis.Results The weight growth rate of the control group was significantly higher than that of the SiO2 dusting group.Compared with the control group,the lung mass and lung coefficient of the SiO2 dusting group were significantly increased(P<0.05).The inflammatory response of the lungs in the control group was significantly reduced at 21 and 28 days,and the inflammatory cells infiltrated in the lung tissue of the SiO2 group.The rats in the control group had a small amount of collagen at 21 and 28 days.A large amount of collagen fiber deposition began to appear in the lung tissue of rats exposed to SiO2 for 21 days.Compared with the control group,the expression levels of miRNA-29b-3p and miRNA-34c-3p in th

关 键 词：矽肺 肺纤维化 微RNA 大鼠 差异表达 

分 类 号：R135.2[医药卫生—劳动卫生]