郭冬芬,任真奎,安小琼,陈明,孙嫚彬,吴昌学,禹文峰
GUO Dongfen,REN Zhenkui,AN Xiaoqiong,CHEN Ming,SUN Manbin,WU Changxue,YU Wenfeng

• 1:贵州医科大学分子生物学重点实验室
• 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
• 3:贵州省第二人民医院检验科
• 4:贵州医科大学儿科系

摘要(Abstract)：

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（Csf2）、IL-1β、丝氨酸（或半胱氨酸）肽酶抑制剂B分支成员2(Serpinb2) mRNA表达增加（P<0.05），而粒细胞集落刺激因子（Csf3）、Cd40抗原（Cd40）、前列腺素内过氧化物合酶2(Ptgs2)、IL-10、CXC基序趋化因子配体2 (Cxcl2)以及IL-12bmRNA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,Csf2,IL-1β,Serpinb2(P< 0. 05);whereas the expression ofCsf3,Cd40,Ptgs2,IL-10,Cxcl2, andIL-12 bdeclined(P< 0. 05).C onclusion 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(KeyWords)： 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
stroke;BV2 cell;glucose oxygen deprivation/reoxygenation;bioinformatics analysis;differentially expressed genes;protein-protein interaction networks

Abstract:

Keywords:

基金项目(Foundation): 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）

作者(Author): 郭冬芬,任真奎,安小琼,陈明,孙嫚彬,吴昌学,禹文峰
GUO Dongfen,REN Zhenkui,AN Xiaoqiong,CHEN Ming,SUN Manbin,WU Changxue,YU Wenfeng

DOI: 10.19367/j.cnki.2096-8388.2022.03.005

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