目的: 探讨缺血性卒中(Ischemic Stroke,IS)患者的循环免疫环状RNA(circular RNA,circRNA)-微小RNA(microRNA,miRNA)-信使RNA(messenger RNA,mRNA)轴,寻找治疗IS的分子靶点。方法: 从GEO数据库中检索到了GSE16561、GSE195442数据集,使用R软件或者GEO2R工具查找缺血性卒中患者与正常对照血液之间差异表达的基因、circRNAs。通过STRING数据库,构建蛋白质-蛋白质相互作用(PPI)网络,用Cytoscape3.9.1进行可视化,使用CytoNCA中的介数中心度筛选出评分较高的基因。对筛选出前15个评分较高的差异基因进行基因本体论(GO)和京都基因和基因组百科全书(KEGG)途径富集分析。之后,根据mirTargets 2.0数据库预测靶标miRNAs,构建miRNA-mRNA调控网络。使用GEO2R工具筛选出IS患者差异表达的circRNAs,结合文献挑选出显著差异并且已经经过PCR验证过的circRNAs,使用Circular RNA Interactome构建circRNA-miRNA调控网络。对两组调控网络取交集,预测出IS中可能存在的调控网络,进一步构建免疫相关circRNA-miRNA-mRNA轴,使用GSE117064数据集对免疫相关circRNA-miRNA-mRNA轴中的miRNAs进行验证。结果: 本研究鉴定了许多转录因子,基因包括CD247、IL2RB、STAT1,miRNAs包括hsa-miR-892a、hsa-miR-938、hsa-miR-338-3p等,并且预测了hsa_circ_0112036 - hsa-miR-892a - CD247、hsa_circ_0112036 - hsa-miR-892a - IL2RB、hsa_circ_0093708 - hsa-miR-1248 - STAT1等7条可能的免疫相关circRNA-miRNA-mRNA轴。结论: hsa_circ_0112036可能作为hsa-miR-892a、hsa-miR-938、hsa-miR-1276、hsa-miR-1208、hsa-miR-338-3p的分子海绵调控IS的进展。hsa_circ_0093708可能作为hsa-miR-1248的分子海绵调控IS的进展。
Objective: To investigate the circulating immune circRNA-miRNA-mRNA axis in patients with ischemic stroke (IS), and to find molecular targets for the treatment of IS. Methods: The GSE16561, GSE195442 datasets were retrieved from the GEO database, using the R software or the GEO2R tool to find the differentially expressed genes ,circRNAs between the blood of ischemic stroke patients and normal controls. A protein-protein interaction (PPI) network was constructed through the STRING database, visualized with Cytoscape 3.9.1, and genes with high scores were selected using the Betweenness Centrality in CytoNCA. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed on the top 15 differential genes with high scores. Thereafter, the miRNA-mRNA regulatory network was constructed by predicting the target miRNAs according to the mirTargets 2.0 database. The differentially expressed circRNAs in IS patients were selected using the GEO2R tool, Combined with literature selected significant differences circRNAs and what has been validated by PCR, the circRNA-miRNA regulatory network was constructed using Circular RNA Interactome. The intersection of the two regulatory networks predicted the possible regulatory networks in the IS, and further constructed the immune-related circRNA-miRNA-mRNA axis, Validation of the miRNAs in the immune-related circRNA-miRNA-mRNA axis using the GSE117064 dataset. Results: This study identified many transcription factors, genes including CD247, IL2RB, STAT1, and miRNAs including hsa-miR-892a, hsa-miR-938, hsa-miR-338-3p, and predicted seven possible immune-related circRNA-miRNA-mRNA axes, for example hsa_circ_0112036 - hsa-miR-892a - CD247, hsa_circ_0112036 - hsa-miR-892a - IL2RB, hsa_circ_0093708 - hsa-miR-1248 - STAT1. Conclusion: The hsa _ circ _ 0112036 may serve as a molecular sponge of hsa-miR-892a, hsa-miR-938, hsa-miR-1276, hsa-miR-1208, and hsa-miR-338-3p to regulate the progression of IS. Regulating IS progression as hsa _ circ _ 0093708 may act as a molecular sponge for hsa-miR-1248.
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