氧糖剥夺再灌注后BV2细胞中差异表达关键基因的筛选及鉴定  

Screening and identification of differential expression genes of BV2 cells induced by oxygen-glucose oxygen deprivation/reoxygenation

作　　者：郭冬芬 任真奎 安小琼 陈明 孙嫚彬 吴昌学[1,2] 禹文峰[1,2] GUO Dongfen;REN Zhenkui;AN Xiaoqiong;CHEN Ming;SUN Manbin;WU Changxue;YU Wenfeng(Laboratory of Molecular Biology,Guizhou Medical University,Guiyang 550004,Guizhou,China;Key Laboratory of Endemic and Minority Diseases of Ministry of Education,Guizhou Medical University,Guiyang 550004,Guizhou,China;Clinical Laboratory,the 2 nd People's Hospital of Guizhou Province,Guiyang 550004,Guizhou,China;Department of Pediatrics,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学分子生物学重点实验室,贵州贵阳550004 [2]贵州医科大学地方病与少数民族疾病教育部重点实验室,贵州贵阳550004 [3]贵州省第二人民医院检验科,贵州贵阳550004 [4]贵州医科大学儿科系,贵州贵阳550004

出　　处：《贵州医科大学学报》2022年第3期273-278,共6页Journal of Guizhou Medical University

基　　金：国家自然科学基金(81360199,82060232);中央引导地方科技科技发展专项资金(〔2019〕4008);贵州省卫生健康委科学技术基金(gzwjkj2019-1-039)。

摘　　要：目的筛选氧糖剥夺再灌注后BV2细胞中差异表达的基因。方法取小鼠小胶质细胞BV2分为正常组(control组)和氧糖剥夺再灌注组(OGD/R组),对两组细胞进行转录测序筛选差异表达基因,在线网站(https://www.xiantao.love/products)对差异表达基因进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)功能富集分析;数据库String(https://cn.string-db.org/)对差异表达基因进行蛋白质互作网络分析后,Cytoscape软件中选择cytoHubba插件筛选出评分较高的关键基因;实时荧光定量PCR(qRT-PCR)检测两组关键基因的mRNA表达水平。结果与control组比较,OGD/R组有122个基因发生明显差异表达(P<0.01),其中108个基因表达上调,14个基因表达下调;GO分析显示,122个差异表达基因与受体配体激活、静脉曲张、正调控STAT信号通路等相关;KEGG分析显示,差异表达基因主要富集到白细胞介素-17(IL-17)信号通路中;蛋白质互作网络分析筛选出差异表达基因中10个评分较高的基因;qRT-PCR结果显示,与control组比较,OGD/R组中IL-6、粒细胞-巨噬细胞集落刺激因子(Csf 2)、IL-1β、丝氨酸(或半胱氨酸)肽酶抑制剂B分支成员2(Serpinb 2)mRNA表达增加(P<0.05),而粒细胞集落刺激因子(Csf 3)、Cd 40抗原(Cd 40)、前列腺素内过氧化物合酶2(Ptgs 2)、IL-10、CXC基序趋化因子配体2(Cxcl 2)以及IL-12 b mRNA表达降低(P<0.05)。结论OGD/R处理可诱导BV2细胞基因差异表达,生物信息学分析可筛选出差异表达基因中的关键基因,qRT-PCR可检测关键基因发生差异表达的情况。Objective Identification of key differential expression genes after glucose oxygen deprivation/reoxygenation in BV2 cell.Methods Rats microglia cell BV2 was divided into control group and OGD/R group;both groups were subjected to RNA-sequencing.Differentially expressed genes(DEGs)of both groups were transcribe screening filtered.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)were performed for the functional and pathway enrichment analyses on the online website(https://www.xiantao.love/products).DEGs were input to the String database(https://cn.string-db.org/)for the protein-protein interaction networks analyze,then,cytoHubba plugin were screed for key genes with higher scores by Cytoscape.The mRNA expression level were validated by qRT-PCR in both control and OGD/R group.Results As comparison with control group,there were 122 differentially expressed genes in OGD/R group(P<0.01).Of which,108 genes were upregulated,14 were down-regulated;GO analysis revealed 122 DEGs correlated with receptor ligand activation,varicose,positive regulation of STAT signal pathway.KEGG results indicated DEGs mainly enriched in the IL-17 signal pathway.Ten key genes with higher evaluation scores were screened via PPI networks;compared with control group,qRT-PCR result showed that the mRNA expression level upregulated:IL-6,Csf 2,IL-1β,Serpinb 2(P<0.05);whereas the expression of Csf 3,Cd 40,Ptgs 2,IL-10,Cxcl 2,and IL-12 b declined(P<0.05).Conclusion OGD/R induced BV2 differentially expressed genes,bioinformatics analysis can screen key genes of such DEGs,and qRT-PCR can detect differentially expressed key genes condition.

关 键 词：脑卒中 BV2细胞 氧糖剥夺再灌注 生物信息学分析 差异表达基因 蛋白质互作网络分析 

分 类 号：R34[医药卫生—基础医学]