摘要
目的基于基因表达数据库(GEO)分析大鼠中暑神经损伤的基因表达差异,探讨lncRNA-miRNA-mRNA共表达调控网络(ceRNA网络)参与中暑神经损伤的作用。方法通过GEO数据库检索中暑相关数据集GSE64778获得差异表达基因(differential expression genes,DEGs),结合GeneCards和CTD数据库检索中暑相关靶基因,取交集筛选获得候选靶基因。采用R语言对中暑相关候选靶基因进行GO和KEGG富集分析;进一步STRING数据库构建靶基因的交互作用网络图,并对关键基因进行相关性分析获得核心基因。通过RNAInter、miRWalk及RAID2数据库对候选靶基因的上游miRNA进行预测,再利用miRDB数据库预测与lncRNA靶向结合的miRNA,筛选并构建lncRNA-miRNA-mRNA共表达调控通路。结果GSE64778数据集的SRA数据筛选,共获得1178个DEGs,其中322个上调,856个下调。GeneCards数据库筛选到2914个基因,CTD数据库筛选到2377个基因。将GSE64778数据集差异表达排名前300的基因,与GeneCards和CTD数据库检索结果取交集,最终筛出25个候选DEGs。GO功能注释结果表明靶基因主要参与细胞凋亡、应激反应以及细胞过程的负调控,在蛋白质二聚化和蛋白结合等过程中发挥功能。KEGG富集分析提示候选靶基因主要富集在PI3K-Akt信号通路。蛋白-蛋白交互作用(PPI)网络分析Hsp90aa1是degree值最高的枢纽基因,且在PI3K-Akt信号通路中富集。对Hsp90aa1基因的上游miRNA以及候选lncRNA的靶miRNA进行预测,锁定3个lncRNAs、8个miRNAs和1个mRNA,DIANA TOOLS数据库通路富集和相关性分析得到LOC102547734与miR-206-3p、miR-206-3p与Hsp90aa1存在靶向结合位点。结论基于生物信息学分析,中暑神经损伤与细胞凋亡、应激反应以及细胞过程的负调控相关;LOC102547734-miR-206-3p-Hsp90aa1调控网络可能抑制中暑神经损伤的发生。
Objective The gene expression differences of heat stroke-induced nerve injury in rats based on the GEO database were analyzed to explore the role of the lncRNA-miRNA-mRNA co-expression regulatory network involved in nerve injury caused by heat stroke.Methods The GEO database was used to retrieve the heat-stroke-related dataset GSE64778 to obtain differentially expressed genes(DEGs),and the GeneCards and CTD databases were combined to retrieve heat-stroke-related target genes,and the candidate target genes were obtained by intersection screening.The R language was used to enrich GO and KEGG for heat-stroke-related candidate target genes.Furthermore,STRING database was used to construct the interaction network map of target genes and correlation analysis of key genes to obtain core genes.Prediction of upstream miRNAs of candidate target genes by the RNAInter,miRWalk,and RAID2 databases,and then prediction of miRNAs that bind to lncRNA targets by the miRDB database to screen and construct a lncRNAmiRNA-mRNA co-expression regulatory network.Results SRA data screening of the GSE64778 dataset yielded 1178 DEGs,of which 322 were up-regulated and 856 were down-regulated.2914 genes were screened from the GeneCards database,and 2377 genes were screened from the CTD database.The top 300 differentially expressed genes in the GSE64778 dataset were intersected with the GeneCards and CTD database search results,and 25 candidate DEGs were finally screened.GO functional annotation results indicated that the target genes were mainly involved in apoptosis,stress response,and negative regulation of cellular processes and functioned in processes such as protein dimerization and protein binding.KEGG enrichment analysis suggested that the candidate target genes were mainly enriched in the PI3K-Akt signaling pathway,and Hsp90aa1 was the pivotal gene with the highest degree value and was enriched in the PI3K-Akt signaling pathway by PPI network analysis.Prediction of upstream miRNAs of the Hsp90aa1 gene and target miRNAs of candidate lncRNAs targeted 3 lncRNAs,8 miRNAs,and 1 mRNA.DIANA TOOLS database pathway enrichment and correlation analysis revealed targeted binding sites between LOC102547734 and miR-206-3p,miR-206-3p and Hsp90aa1.Conclusions Based on bioinformatics analysis,we found that heat-shot-induced nerve injury is associated with apoptosis,stress response,and negative regulation of cellular processes.The LOC102547734-miR-206-3p-Hsp90aa1 regulatory network may inhibit the occurrence of heat-shot nerve injury.
作者
王蕾
姜岱山
朱保锋
贾寒雨
沈君华
张毅
WANG Lei;JIANG Daishan;ZHU Baofeng;JIA Hanyu;SHEN Junhua;ZHANG Yi(Emergency Center of the Second Affiliated Hospital of Nantong University,Nantong,Jiangsu 226001,China;Emergency Center of the Affiliated Hospital of Nantong University,Nantong,Jiangsu 226001,China;Department of Science and Education,the Second Affiliated Hospital of Nantong University,Nantong,Jiangsu 226001,China)
出处
《职业卫生与应急救援》
2023年第5期623-630,共8页
Occupational Health and Emergency Rescue
基金
江苏省卫生健康委员会医学研究项目(Z2022067)
江苏省“六大人才高峰”高层次人才项目(2019-WSW-199)
南通市卫健委课题(MB2021018、MB2021026)。
