儿童哮喘基因表达谱芯片的生物信息学分析  

Bioinformatics analysis of gene expression profiling chips in childhood asthma

作　　者：苏保宁 马爱玲 赵文慧 SU Baoning;MA Ailing;ZHAO Wenhui(Department of Pediatrics,Shanghai Jiading District Hospital of Traditional Chinese Medicine,Shanghai,201800,China;Department of Internal Medicine,Shanghai Jiading District Hospital of Traditional Chinese Medicine,Shanghai,201800,China;Department of Clinical Laboratory,Affiliated to Xuzhou Medical University Lianyungang Oriental Hospital,Lianyungang,222042,Jiangsu,China)

机构地区：[1]上海市嘉定区中医医院儿科,上海201800 [2]上海市嘉定区中医医院内科,上海201800 [3]徐州医科大学附属连云港市立东方医院检验科,江苏连云港222042

出　　处：《当代医学》2022年第28期1-7,共7页Contemporary Medicine

基　　金：上海市中医专科培育项目[ZY﹝2019-2020﹞-ZYZK-34];上海市优势病种建设项目[ZY﹝2018-2020﹞-ZYBZ-40];嘉定卫健委科研项目(2018-KY-ZYY-11)。

摘　　要：目的应用生物信息学分析方法对儿童哮喘(CA)基因表达谱芯片进行分析,从分子和细胞水平探讨CA发病机制。方法从基因表达综合数据库(GEO)下载CA基因芯片数据集GSE16032,利用生物信息学方法筛选差异表达基因(DEGs),并利用注释可视化和集成发现数据库(DAVID)分析工具对DEGs进行基因本体论(GO)及信号通路富集(KEGG)分析,通过STRING在线软件、Cytoscape及其插件MCODE对DEGs进行蛋白质-蛋白质相互作用网络分析(PPI分析)。利用CIBERSORT在线数据库分析样本外周血中免疫细胞的免疫浸润变化情况,实时荧光定量(qRT-PCR)验证关键基因的表达情况。结果本研究共筛选出244个DEGs,包括233个上调基因和11个下调基因。GO功能和KEGG信号结果显示,DEGs主要参与炎症和免疫反应及细胞黏附等生物过程,涉及血小板激活、细胞因子和细胞因子受体互作及Ras、TGF-β等信号通路。通过STRING和Cytoscape软件分析发现G蛋白γ11亚基(GNG11)、人琥珀酸受体1(SUCNR1)、CXC趋化因子配体5(CXCL5)、促血小板碱性蛋白(PPBP)、人源嘌呤能受体12(P2RY12)、甲酰肽受体1(FPR1)、趋化因子受体2(CCR2)等13个关键基因。免疫细胞浸润分析显示,不同样本中免疫细胞分布比例具有差异性。与哮喘急性期相比,哮喘恢复期免疫细胞中的单核细胞比例下降,而CD8+T细胞和静息自然杀伤细胞(NK细胞)比例均上升。qRT-PCR结果进一步验证了13个关键基因的变化水平,12个关键基因在哮喘恢复期外周血中比例下降。结论包括GNG11在内的13个关键基因和免疫细胞浸润在CA的发生与发展中起重要作用,可为研究CA的发病机制提供新的思路和治疗策略。Objective To analyze the gene expression microarray of childhood asthma(CA)by bioinformatics analysis to explore the pathogenesis of CA at the molecular and cellular levels.Methods Download the CA gene chip dataset GSE16032 from the gene expression omnibus(GEO),screen differentially expressed genes(DEGs)by bioinformatics method,and perform gene ontology(GO)and kyoto encyclopedia of genes and genomes(KEGG)analysis of DEGs using annotated visualization and integrated discovery database(DAVID)analysis tools.Cytoscape and its plugin MCODE perform protein-protein interaction network analysis(PPI analysis)on DEGs.CIBERSORT online database was used to analyze the changes of immune infiltration of immune cells in the peripheral blood of samples,and Quantitative Real-time PCR(qRT-PCR)to verify the expression of key genes.Results A total of 244 DEGs were screened out,including 233 up-regulated genes and 11 down-regulated genes.GO function and KEGG signaling results showed that DEGs were mainly involved in biological processes such as inflammation,immune response and cell adhesion,involving platelet activation,cytokine and cytokine receptor interaction,and signaling pathways such as Ras and TGF-β.Through STRING and Cytoscape software analysis,13 key genes such as G proteinγ11 subunits(GNG11),human succinic acid receptor 1(SUCNR1),CXC chemokine ligand 5(CXCL5),pro-platelet basic protein(PPBP),human purineergic receptor 12(P2RY12),formyl peptide receptor 1(FPR1),chemokine receptor 2(CCR2)were found.Immune cell infiltration analysis showed that the proportion of immune cells distributed in different samples was different.Compared with the acute phase of asthma,the proportion of monocytes in immune cells convalescent from asthma decreased,while the proportion of CD8+T cells and resting natural killer cells(NK cells)increased.The qRT-PCR results further verified the level of change in 13 key genes,and the proportion of 12 key genes in peripheral blood during asthma recovery was reduced.Conclusion The infiltration of 13 key gen

关 键 词：儿童哮喘 基因表达谱芯片 差异表达基因 生物信息学 

分 类 号：R725.6[医药卫生—儿科]