脓毒症急性肾损伤关键基因的筛选和验证研究

Screening and Validation of Key Genes in Sepsis-Associated Acute Kidney Injury

摘要: 目的：筛选脓毒症急性肾损伤(sepsis-associated acute kidney injury, SA-AKI)相关的关键基因，分析其与临床结局的相关性，并通过对关键基因进行验证。方法：纳入GEO数据库中脓毒症、急性肾损伤(AKI)相关转录组数据集，采用批次校正和差异分析方法筛选共同差异表达基因(DEGs)，并进行GO和KEGG富集分析。进一步分析关键基因在健康对照与脓毒症患者、存活者及非存活者中的表达差异，筛选与临床结局相关的核心基因。采用盲肠结扎穿孔(CLP)法建立小鼠脓毒症模型，通过HE染色观察肾组织病理变化，ELISA检测肾组织中IL-1β、IL-6、IL-10和TNF-α水平，qPCR验证核心基因表达。结果：差异分析共筛选出11个与SA-AKI相关的差异表达基因，功能富集分析显示其主要涉及肽酶调节活性、免疫应答调控等生物学过程。进一步分析发现，MS4A4A、VSIG4、CKLF、LY96、CD163和CSTA共6个基因在脓毒症患者中显著高表达，且在非存活者中的表达水平进一步升高。动物实验结果显示，与对照组相比，CLP组小鼠肾组织病理损伤明显，IL-1β、IL-6、IL-10和TNF-α水平显著升高，且上述6个基因在肾组织中的mRNA表达均明显上调。结论：本研究筛选并验证了6个与SA-AKI临床结局相关的关键基因。MS4A4A、VSIG4、CKLF、LY96、CD163和CSTA可能通过参与炎症反应及肾组织损伤过程促进SA-AKI的发生发展，有望成为评估疾病严重程度及不良结局的潜在分子标志物。

Abstract: Objective: To identify key genes associated with sepsis-associated acute kidney injury (SA-AKI), analyze their correlation with clinical outcomes, and validate these key genes. Methods: Transcriptomic datasets related to sepsis and acute kidney injury (AKI) were obtained from the GEO database. Batch effect correction and differential expression analysis were performed to identify common differentially expressed genes (DEGs), followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The expression differences of key genes were further analyzed between healthy controls and sepsis patients, as well as between survivors and non-survivors, to screen core genes associated with clinical outcomes. A mouse sepsis model was established by cecal ligation and puncture (CLP). Histopathological changes in renal tissues were observed by hematoxylin-eosin (HE) staining. The levels of IL-1β, IL-6, IL-10, and TNF-α in renal tissues were measured by ELISA, and the expression of core genes was validated by quantitative polymerase chain reaction (qPCR). Results: A total of 11 differentially expressed genes associated with SA-AKI were identified. Functional enrichment analysis showed that these genes were mainly involved in biological processes such as regulation of peptidase activity and immune response regulation. Further analysis revealed that six genes, namely MS4A4A, VSIG4, CKLF, LY96, CD163, and CSTA, were significantly upregulated in sepsis patients, and their expression levels were further increased in non-survivors. Animal experiments showed that, compared with the control group, the CLP group exhibited obvious renal histopathological injury, significantly increased levels of IL-1β, IL-6, IL-10, and TNF-α, and markedly upregulated mRNA expression of the above six genes in renal tissues. Conclusion: This study identified and validated six key genes associated with the clinical outcomes of SA-AKI. MS4A4A, VSIG4, CKLF, LY96, CD163, and CSTA may promote the occurrence and progression of SA-AKI by participating in inflammatory responses and renal tissue injury, and may serve as potential molecular biomarkers for evaluating disease severity and poor clinical outcomes.