基于转录组测序的煤工尘肺大鼠模型中线粒体相关差异基因筛选及分析  

Screening and Analysis of Mitochondria-related Differential Genes in Rat Model of Coal Workers’Pneumoconiosis Based on Transcriptome Sequencing

作　　者：刘道伟 张娜[1,2] 刘琭璐 章梅 杨惠芳 LIU Daowei;ZHANG Na;LIU Lulu;ZHANG Mei;YANG Huifang(School of Public Health,Ningxia Medical University,Yinchuan 750004,China;Ningxia Key Laboratory of Environmental Factors and Chronic Disease Control,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学公共卫生学院,银川750004 [2]宁夏环境因素与慢性病控制重点实验室,银川750004

出　　处：《宁夏医科大学学报》2024年第1期21-25,31,共6页Journal of Ningxia Medical University

基　　金：宁夏回族自治区重点研发计划项目(2023BEG02028);国家自然科学基金项目(82160608);宁夏自然科学基金项目(2022AAC03141,2021AAC02013);宁夏回族自治区重点研发计划一般项目(2021BEB04059)。

摘　　要：目的应用高通量转录组测序技术,对煤工尘肺大鼠模型中线粒体相关差异基因进行筛选及分析。方法选取无特定病原体(SPF)级SD雄性大鼠30只,根据随机数表法分为对照组、煤尘组、矽尘组,每组10只,采用一次性非暴露式气管内灌注法造模,并常规饲养90 d。取肺组织制作病理标本,观察其病理改变,采用Masson染色观察其纤维化情况,免疫组化检测肺组织的胶原蛋白Ⅰ(COL-Ⅰ)表达情况来确定造模情况。利用转录组测序技术对样本间基因进行差异表达分析,筛选出差异表达基因,然后对差异基因进行GO和KEGG生物信息学分析,进一步筛选出大鼠煤工尘肺模型中与线粒体有关的差异基因,并进行生物信息学分析。结果HE、Masson染色及免疫组化结果显示,煤尘组与矽尘组中COL-Ⅰ表达量增高。对对照组与煤尘组的差异基因进行筛选,其中与线粒体有关的基因一共35个(上调28个和下调7个)。对对照组与矽尘组的差异基因进行筛选,其中与线粒体有关的基因一共302个(上调202个和下调100个)。通过GO富集分析发现,在生物过程中,细胞过程、单一的有机体过程较为重要;在分子功能中,以活动分子传感器、催化活性为主;在细胞组分中,以细胞部分、细胞为主。对照组与矽尘组中与线粒体相关的差异基因通过KEGG显著富集到前20条通路中,氧化磷酸化、产热、帕金森综合征、朊病毒病等通路富集到的基因最多。结论通过转录组高通量测序,筛选分析了煤工尘肺大鼠模型中线粒体相关差异基因,以期为后续探索煤工尘肺与线粒体的关系提供依据。Objective To screen and analyze mitochondria-related differential genes in the rat model of coal workers’pneumoconiosis by high-throughput transcriptome sequencing.Methods Thirty male SD rats free of specific pathogen(SPF)were selected and divided into the control group,the coal dust group and the silica dust group according to the principle of randomization.The rats were molded by one-time non-exposure endotracheal perfusion and fed for 90 days.The pathological specimens of lung tissue were taken and the pathological changes were observed.The fibrosis was observed by Masson staining.The expression of collagenousⅠ(COL-Ⅰ)in lung tissue was detected by immunohistochemistry to confirm the success of modeling.Then,transcriptome sequencing technology was used to analyze the differential expression of genes among the samples and screen out the differentially expressed genes.GO and KEGG bioinformatics analysis were performed on the differentially expressed genes,and the differentially expressed genes related to mitochondria in the rats model of coal workers’pneumoconiosis were further screened out and bioinformatics analysis was conducted.Results The HE、Masson and ELISA showed that in the coal dust group and silica dust group,COL-Ⅰincreased significantly.There were 35 genes related to mitochondria(including 28 up-regulated and 7 down-regulated)in the control group and the coal dust group.The differential genes of the control group and the silica dust group were screened,and a total of 302 genes related to mitochondria(including 202 up-regulated and 100 down-regulated)were detected.Through GO enrichment analysis,it was found that cellular process and single biological process were more important in biological process.In the molecular function,the active molecular sensor and catalytic activity were the main.Among the cell components,cell parts and cells were the main ones.Among the top 20 pathways in which mitochondria-related differential genes in the control group and the dust group were significantly enrich

关 键 词：煤工尘肺 转录组测序 线粒体 肺纤维化 

分 类 号：R135.2[医药卫生—劳动卫生]