急性缺血性卒中相关基因的筛选和验证  

Screening and verification of genes related to ischemic stroke

作　　者：蒋蔚 万蛟龙 包元飞[2] 王新 JIANG Wei;WAN Jiaolong;BAO Yuanfei;WANG Xin(Brain Center,Wuxi Second People's Hospital,Nanjing Medical University,Wuxi Jiangsu 214000;Department of Neurology,Nantong Third People's Hospital,Nantong University,Nantong Jiangsu 226001;Jiangsu Key Laboratory of Nerve Regeneration,Nantong Jiangsu 226001,China)

机构地区：[1]南京医科大学附属无锡市第二人民医院脑科中心,江苏无锡214000 [2]南通大学附属南通第三人民医院神经内科,江苏南通226001 [3]江苏省神经再生重点实验室,江苏南通226001

出　　处：《临床与病理杂志》2020年第9期2325-2331,共7页Journal of Clinical and Pathological Research

基　　金：南通市自然科学基金(MS12017021-4)。

摘　　要：目的:通过分析基因表达谱,探讨缺血性脑卒中(ischemic stroke,IS)的分子机制和诊断标志物。方法:下载GEO数据库中缺血性卒中患者和对照组的基因芯片数据,使用Pubmed提供的GEO2R软件和R语言筛选出具有显著意义的差异基因(differential genes,DEGs);利用Metascape进行基因本体(gene ontology,GO)与酶和通路数据库(Reactome)基因集分析;使用STRING在线分析工具和Cytoscape软件分析出关键基因;最后,收集急性卒中患者外周血,通过RT-PCR进行验证。结果:通过分析IS患者外周血基因芯片数据集,共筛选出20个差异表达基因;基因注释和信号途径分析表明,这些差异表达基因主要参与中性粒细胞脱颗粒、对外来生物刺激反应、细胞因子生产、有机阴离子运输、对脂肪酸的反应和中性粒细胞迁移等。使用Cytoscape和MCODE插件进一步分析,筛选出5个关键基因(MMP-9,OLFM4,ORM1,FOLR3和ARG1);RT-PCR证实ARG1和MMP9 mRNA在IS患者外周血中过表达。结论:缺血性卒中伴随多个基因表达异常,本研究为探索IS的发病机制和开发新的诊治疗策略提供了线索。Objective:The objective of this study was to provide insight into the molecular mechanisms of acute ischemic stroke(IS)through gene expression profiling analysis.Methods:The gene chip data of ischemic stroke patients and the control group in the GEO database were downloaded.Differential genes were analyzed by GEO2R software and the R language.Metascape was used to analyze gene ontology(GO)and gene sets of enzyme and pathway analysis(Reactome).The hub genes were identified using the STRING online analysis tool and Cytoscape software.Finally,quantitative real‐time polymerase chain reaction(qRT‐PCR)was performed to verify the expression of DEGs in the peripheral blood of IS patients.Results:A 20-gene profile was identified in the whole blood of ischemic stroke patients from the gene expression datasets.Gene annotation and signal pathway analysis showed that these differentially expressed genes were mainly involved in the response of neutrophils to extraneous biological stimulation,the response of cytokines to organic anion production,transport to fatty acids,and the migration of neutrophils,etc.Five genes(mmp-9,OLFM4,ORM1,FOLR3,and ARG1)were identified as hub gene by Cytoscape and MCODE software.Finally,ARG1 and MMP9 mRNA overexpressed in peripheral blood of IS patients compared with normal control confirmed by RT-PCR.Conclusion:The dysregulation of several genes may involve the process of ischemic stroke(IS).Our finding provided clues for exploring mechanisms and developing novel diagnostic and therapeutic strategies for IS.

关 键 词：缺血性卒中 精氨酸酶-1 基质金属蛋白酶-9 

分 类 号：R743.3[医药卫生—神经病学与精神病学]