缺血性脑卒中缺氧免疫关键基因的鉴别及诊断价值分析  

Identification and diagnostic value of key genes with hypoxia-immune-based signature in ischemic stroke

作　　者：黄小兰 杨佳磊 梁天 贺婉婷 吕妙 刘声莹 龙建雄[1] 苏莉[1] Huang Xiaolan;Yang Jialei;Liang Tian;He Wanting;Lyu Miao;Liu Shengying;Long Jianxiong;Su Li(School of Public Health,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学公共卫生学院,南宁530021

出　　处：《广西医科大学学报》2023年第8期1274-1282,共9页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81874395,No.81860822);广西壮族自治区自然科学基金资助项目(No.2017GXNSFDA198013,No.2018GXNSFAA281224)。

摘　　要：目的:研究缺血性脑卒中(IS)缺氧免疫机制的相关差异表达基因和调控信号通路,以识别IS潜在诊断生物标志物。方法:从GEO数据库下载IS表达谱数据集GSE58294,采用单样本GSEA(ssGSEA)和t分布随机邻域嵌入算法(t-SNE)评估受试者的缺氧和免疫状态,使用“limma”软件包筛选差异基因(DEGs),通过DAVID在线数据库对DEGs进行基因本体(GO)富集分析及京都基因与基因组百科全书(KEGG)信号通路分析,LASSO筛选与IS关键(hub)基因,然后运用实时荧光定量聚合酶链式反应(RT-qPCR)验证hub基因的差异性表达。结果:筛选出60个IS缺氧免疫DEGs,KEGG结果显示IS缺氧免疫DEGs富集在PI3K-Akt通路,LASSO筛出6个关键基因:CHPF2、MBOAT2、IL18RAP、TIMM8A、ALDOAP2、LPAR4。CHPF2、MBOAT2和IL18RAP为上调基因(均P<0.001),而TIMM8A、ALDOAP2和LPAR4为下调基因(均P<0.001),ROC曲线显示该6个关键基因的AUC为0.8941~0.9943。RT-qPCR结果显示,短暂性大脑中动脉闭塞(tMCAO)小鼠中MBOAT2和IL18RAP的表达显著高于假手术小鼠(均P<0.001)。结论:IL18RAP和MBOAT2可能为IS缺氧免疫相关的DEGs,并可作为IS的潜在诊断生物标志物。Objective:To study the differentially expressed genes and regulatory signaling pathways related to hypoxia and immune mechanisms in ischemic stroke(IS),so as to identify potential biomarkers for the diagnosis of IS.Methods:The IS expression profile dataset GSE58294 was downloaded from the GEO database.Singlesample GSEA(ssGSEA)and t-distributed stochastic neighbor embedding(t-SNE)algorithms were employed to estimate the immune and hypoxia statuses of subjects.The"limma"software package was used to screen differentially expressed genes(DEGs).DAVID online database was used for gene ontology(GO)enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway analysis,least absolute shrinkage and selection operator(LASSO)was utilized to screen IS key(hub)genes,and then real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to verify the differential expressions of key genes.Results:60 hypoxia-immune-IS-related DEGs were screened.KEGG results revealed that the DEGs were enriched in PI3K-Akt signaling pathway.6 key genes-CHPF2,MBOAT2,IL18RAP,TIMM8A,ALDOAP2,and LPAR4 were obtained using the LASSO regression.CHPF2,MBOAT2,and IL18RAP were up-regulated(all P<0.001),while TIMM8A,ALDOAP2 and LPAR4 were down-regulated(all P<0.001).The ROC curve showed that the AUC of these 6 key genes ranged from 0.8941 to 0.9943.RT-qPCR results showed that the expressions of MBOAT2 and IL18RAP in transient middle cerebral artery occlusion(tMCAO)mice were significantly higher than those in sham operation mice(all P<0.001).Conclusion:IL18RAP and MBOAT2 may be hypoxia-immune-IS-related DEGs and may serve as potential biomarkers for the diagnosis of IS.

关 键 词：缺血性脑卒中 缺氧 免疫 MBOAT2 IL18RAPT 

分 类 号：R722[医药卫生—儿科]