脓毒症相关肠黏膜屏障损伤关键基因和信号通路的转录组学分析

Transcriptomic Analysis of Key Genes and Signaling Pathways in Sepsis-Associated Intestinal Mucosal Barrier Damage

目的:分析脓毒症相关肠黏膜屏障损伤小鼠差异表达基因(DEG),从转录组水平探讨脓毒症相关肠黏膜屏障损伤的早期诊断和保护机制。方法:收集行盲肠结扎穿孔术(CLP)和假手术的健康雄性C57BL/6J小鼠小肠组织,采用illumina Hiseq测序平台的两端测序模式进行高通量测序。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)分析。利用STRING数据库构建蛋白–蛋白相互作用(PPI)网络,利用Cytoscape筛选关键基因。采用实时荧光定量聚合酶链反应(RT-qPCR)对Hub基因进行验证。结果:共筛选出239个DEG,其中上调基因49个,下调基因130个。KEGG富集分析发现DEG主要涉及细胞因子–细胞因子受体相互作用、Th1和Th2细胞分化、病毒蛋白与细胞因子和细胞因子受体相互作用、IL-17信号通路等。利用cytoHubba插件筛选出前10个hub基因。经实验验证,TBX21、CSF3、IL-6、CXCR3、CXCL9的表达与测序结果一致。结论:TBX21、CSF3、IL-6、CXCR3和CXCL9可能是脓毒症相关肠黏膜屏障早期诊断和治疗的潜在生物学标志物。

Objectives: To analyze the differentially expressed genes (DEG) in mice with sepsis-related intestinal mucosal barrier damage and to explore the early diagnosis and protection mechanism of sepsis-related intestinal mucosal barrier damage at the transcriptome level. Methods: The healthy male C57BL/6J mice small intestinal tissues of Cecal ligation and puncture (CLP) and sham-operation were collected, and the two-end sequencing mode of the illumina Hiseq sequencing platform was commissioned for high-throughput sequencing. Followed by Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of DEGs. A protein-protein interaction (PPI) network was constructed using the STRING database, and hub genes were selected using Cytoscape. Hub genes were verified using quantitative real-time polymerase chain reaction (RT-qPCR). Results: A total of 239 DEG were selected, including 49 up-regulated genes and 130 downregulated genes. KEGG enrichment analysis revealed that DEG mainly involved cytokine-cytokine receptor interaction, Th1 and Th2 cell differentiation, viral protein interaction with cytokine and cytokine receptor, and IL-17 signaling pathway. The cyto Hubba plugin was used to select the top 10 hub genes. After experimental verification, the expression of TBX21, CSF3, IL-6, CXCR3, and CXCL9 was in agreement with the sequencing results. Conclusion: TBX21, CSF3, IL-6, CXCR3, and CXCL9 may be potential biological markers for the early diagnosis and treatment the sepsis-associated intestinal mucosal barrier.