基于生物信息学分析筛选矽肺患者肺泡巨噬细胞中线粒体靶向标志物  

Screening of mitochondria-targeting markers in alveolar macrophages of silicosis patients based on bioinformatics analysis

作　　者：程鸿明 何海兰 王袁 郝小惠 王宏丽 刘和亮 Cheng Hongming;He Hailan;Wang Yuan;Hao Xiaohui;Wang Hongli;Liu Heliang(School of Public Health,North China University of Science and Technology,Tangshan 063210;Hebei Key Laboratory of Organ Fibrosis,Tangshan 063210;Affiliated Hospital of North China University of Science and Technology,Tangshan 063000;Tangshan Vocational and Technical College,Tangshan 063300)

机构地区：[1]华北理工大学公共卫生学院,唐山063210 [2]河北省器官纤维化重点实验室,唐山063210 [3]华北理工大学附属医院,唐山063000 [4]唐山职业技术学院,唐山063300

出　　处：《安徽医科大学学报》2024年第10期1828-1834,1841,共8页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金项目(编号:U21A20334);河北省自然科学基金项目(编号:H2022209021、H2022209039)。

摘　　要：目的筛选矽肺患者肺泡巨噬细胞中线粒体靶向差异基因,探究线粒体稳态在矽肺患者肺泡巨噬细胞中的作用。方法下载高通量测序数据集GSE174725,使用R软件以P<0.05,|log 2(Fold Change)|>1筛选差异表达基因,将其与线粒体基因库MitoCarta3.0取交集得到线粒体靶向差异表达基因,再开展富集分析得到差异基因参与的生物过程和通路,构建差异表达基因蛋白质-蛋白质相互作用网络。此外,收集矽肺患者对照人群肺泡巨噬细胞,利用RT-qPCR法检测差异基因的表达水平以及利用Western blot探究线粒体相关因子线粒体融合蛋白-1(MFFN1)、视神经萎缩蛋白-1(OPA1)、动力蛋白相关蛋白-1(DRP1)在矽肺患者肺泡巨噬细胞中的表达情况。结果在矽肺患者中共筛选出差异表达基因204个,包括上调差异基因62个,下调差异基因142个,其中线粒体靶向差异表达基因22个。线粒体靶向差异表达基因富集分析显示,其主要富集的细胞组成包括线粒体膜、内质网膜管侧的组成部分等,主要富集的生物过程包括线粒体电子传递还原型烟酰胺腺嘌呤二核苷酸到泛醌、炎症反应、免疫反应等,主要富集的分子功能包括质子转运腺嘌呤核苷三磷酸合酶活性旋转机制、NADH脱氢酶活性、趋化因子活动等,京都基因与基因组百科全书富集分析主要集中在参与化学致癌-活性氧、白细胞介素17信号通路、toll样受体信号通路、趋化因子信号通路、肿瘤坏死因子信号通路等。此外,RT-qPCR结果显示,矽肺患者中线粒体细胞色素C氧化酶1、线粒体细胞色素C氧化酶2、线粒体细胞色素C氧化酶3、线粒体编码的NADH脱氢酶1、线粒体编码的NADH脱氢酶3、线粒体编码的NADH脱氢酶5、超氧化物歧化酶和线粒体编码的ATP合酶6表达下调(P<0.05),Western blot和RT-qPCR结果显示,矽肺患者中MFN1、OPA1表达降低(P<0.05),DRP1表达增高(P<0.05)。结论经生物信息学分析�Objective To screen mitochondria-targeting differential genes in alveolar macrophages of silicosis patients and explore the role of mitochondrial homeostasis in alveolar macrophages of silicosis patients.Methods High-throughput sequencing dataset GSE174725 was downloaded,and differentially expressed genes were screened with R software and P<0.05,|LogFC|>1,and then intermixed with mitochondrial gene bank MitoCarta3.0 to obtain mitochondria-targeted differentially expressed genes.Then enrichment analysis was carried out to obtain the biological processes and pathways involved in differential genes,and the protein-protein interaction network was constructed.In addition,alveolar macrophages from silicosis patients and healthy controls were collected,the expression levels of differential genes were detected by RT-qPCR,and the expressions of mitochondria-related factors mitochondrial fusion protein 1(MFN1),optic atrophy 1(OPA1)and dynamin-related protein 1(DRP1)in alveolar macrophages of silicosis patients were investigated by Western blot.Results A total of 204 differentially expressed genes were screened in silicosis patients,among which 62 differentially expressed genes were up-regulated,142 differentially expressed genes were down-regulated,and 22 differentially expressed genes were mitochondria-targeted.The concentration analysis of differentially expressed genes targeted by mitochondria showed that the cell components mainly enriched included mitochondrial membrane,endoplasmic membrane side components,etc.The biological processes mainly enriched included mitochondrial electron transfer from NADH to ubiquinone,inflammatory response,immune response,etc.The main molecular functions enriched included the rotation mechanism of proton transport ATP synthase activity,NADH dehydrogenase activity,chemokine activity,etc.KEGG enrichment analysis mainly focused on the involvement in chemical carcinogenesis-ROS,IL-17 signaling pathway,toll-like receptor signaling pathway,chemokine signaling pathway,TNF signaling pathway,etc.I

关 键 词：矽肺 巨噬细胞 线粒体分裂 线粒体融合 生物信息学 

分 类 号：R135.2[医药卫生—劳动卫生]