基于生物信息学分析鉴定急性心肌梗死中与糖酵解相关的枢纽基因及其与免疫浸润的相关性。

PURPOSE: Glycolysis and immune metabolism play important roles in acute myocardial infarction (AMI). Therefore, this study aimed to identify and experimentally validate the glycolysis-related hub genes in AMI as diagnostic biomarkers, and further explore the association between hub genes and immune infiltration. METHODS: Differentially expressed genes (DEGs) from AMI peripheral blood mononuclear cells (PBMCs) were analyzed using R software. Glycolysis-related DEGs (GRDEGs) were identified and analyzed using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) for functional enrichment. A protein-protein interaction network was constructed using the STRING database and visualized using Cytoscape software. Immune infiltration analysis between patients with AMI and stable coronary artery disease (SCAD) controls was performed using CIBERSORT, and correlation analysis between GRDEGs and immune cell infiltration was performed. We also plotted nomograms and receiver operating characteristic (ROC) curves to assess the predictive accuracy of GRDEGs for AMI occurrence. Finally, key genes were experimentally validated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting using PBMCs. RESULTS: A total of 132 GRDEGs and 56 GRDEGs were identified on the first day and 4-6 days after AMI, respectively. Enrichment analysis indicated that these GRDEGs were mainly clustered in the glycolysis/gluconeogenesis and metabolic pathways. Five hub genes (HK2, PFKL, PKM, G6PD, and ALDOA) were selected using the cytoHubba plugin. The link between immune cells and hub genes indicated that HK2, PFKL, PKM, and ALDOA were significantly positively correlated with monocytes and neutrophils, whereas G6PD was significantly positively correlated with neutrophils. The calibration curve, decision curve analysis, and ROC curves indicated that the five hub GRDEGs exhibited high predictive value for AMI. Furthermore, the five hub GRDEGs were validated by RT-qPCR and western blotting. CONCLUSION: We concluded that HK2, PFKL, PKM, G6PD, and ALDOA are hub GRDEGs in AMI and play important roles in AMI progression. This study provides a novel potential immunotherapeutic method for the treatment of AMI.

目的：糖酵解和免疫代谢在急性心肌梗死（AMI）中发挥重要作用。因此，本研究旨在鉴定和实验验证 AMI 中与糖酵解相关的枢纽基因作为诊断生物标志物，并进一步探讨枢纽基因与免疫浸润的关系。

方法：使用 R 软件分析 AMI 外周血单核细胞（PBMCs）中的差异表达基因（DEGs）。使用数据库 for Annotation, Visualization, and Integrated Discovery（DAVID）鉴定和分析与糖酵解相关的 DEGs（GRDEGs），进行功能富集分析。使用 STRING 数据库构建蛋白质-蛋白质相互作用网络，并使用 Cytoscape 软件可视化。使用 CIBERSORT 分析 AMI 患者和稳定型冠状动脉疾病（SCAD）对照之间的免疫浸润，并对 GRDEGs 与免疫细胞浸润进行相关性分析。我们还绘制了列线图和接收者操作特征（ROC）曲线，以评估 GRDEGs 对 AMI 发生的预测准确性。最后，使用 PBMCs 进行逆转录-定量聚合酶链反应（RT-qPCR）和 Western blot 实验验证关键基因。

结果：在 AMI 发生后第 1 天和第 4-6 天分别鉴定出 132 个和 56 个 GRDEGs。富集分析表明，这些 GRDEGs 主要聚类在糖酵解/糖异生和代谢途径中。使用 cytoHubba 插件选择了 5 个枢纽基因（HK2、PFKL、PKM、G6PD 和 ALDOA）。免疫细胞与枢纽基因之间的联系表明，HK2、PFKL、PKM 和 ALDOA 与单核细胞和中性粒细胞呈显著正相关，而 G6PD 与中性粒细胞呈显著正相关。校准曲线、决策曲线分析和 ROC 曲线表明，这 5 个枢纽 GRDEGs 对 AMI 具有较高的预测价值。此外，通过 RT-qPCR 和 Western blot 验证了这 5 个枢纽 GRDEGs。

结论：我们得出结论，HK2、PFKL、PKM、G6PD 和 ALDOA 是 AMI 中的枢纽 GRDEGs，在 AMI 进展中发挥重要作用。本研究为 AMI 的治疗提供了一种新的潜在免疫治疗方法。