背景:研究的重点是受中性粒细胞胞外诱捕网(NETs)影响的心肌梗死(MI)的长期预后。它还旨在分析和验证此过程中的相对枢纽基因,为了进一步探索能够改善MI预后的新的治疗靶点。
方法:我们通过结扎左前降支(LAD)建立了小鼠MI模型,并进行了8周的连续观察,以研究这些小鼠心脏结构和功能的动态变化。同时,我们服用了Apocynin,NADPH氧化酶的抑制剂,它也被证明抑制了NET的形成,与接受MI手术的小鼠进行比较。本研究采用苏木精-伊红(HE)染色,超声心动图,免疫荧光,和实时定量PCR(RT-qPCR)来检查NETs对MI长期预后的影响。接下来,从GEO数据库下载与MI和NET相关的数据集,分别。R软件的Limma软件包用于鉴定差异表达基因(DEGs)。在分析了“稳健排名聚合(RRA)”包之后,我们对稳健差异表达基因(DEGs)进行了筛选,并使用基因本体论(GO)和京都基因和基因组百科全书(KEGG)进行了通路富集分析,以确定这些稳健DEGs的功能作用.可视化蛋白质-蛋白质相互作用(PPI)网络,并使用Cytoscape过滤hub基因。
结果:免疫荧光和qPCR结果显示,MI后第1周和第8周小鼠心脏中髓过氧化物酶(MPO)的表达增加。HE染色显示MI组心脏在8周内出现一系列病理表现,包括放大的尺寸,心肌细胞排列紊乱,炎症细胞浸润,和过度的胶原纤维沉积,在其他人中。Apocynin的利用可以显着改善这些不良性能。超声心动图显示小鼠心脏的心功能。MI组的心脏运动范围减小,弹射能力下降。此外,发现心室收缩运动异常,随着时间的推移,其壁增厚率下降,表明心肌缺血的进行性恶化。Apocynin组,相反,在上述方面显示出较少的异常变化。共有81个DEGs和4个hub基因(FOS,获得EGR1、PTGS2和HIST1H4H)。RT-qPCR结果显示这4个基因在MI组中表达异常,这可以通过Apocynin的治疗来逆转。
结论:NETs的形成可能与MI高度相关,MI的长期预后可能受到NETs的影响。四个枢纽基因,即FOS,EGR1、PTGS2和HIST1H4H,有可能成为与此过程相关的关键基因。它们还可以作为预测MI预后的生物标志物和作为基因治疗的靶标。
BACKGROUND: The study focuses on the long-term prognosis of myocardial infarction (MI) influenced by neutrophil extracellular traps (NETs). It also aims to analyze and validate relative hub genes in this process, in order to further explore new therapeutic targets that can improve the prognosis of MI.
METHODS: We established a MI model in mice by ligating the left anterior descending branch (LAD) and conducted an 8-week continuous observation to study the dynamic changes in the structure and function of the heart in these mice. Meanwhile, we administered Apocynin, an inhibitor of NADPH Oxidase, which has also been shown to inhibit the formation of NETs, to mice undergoing MI surgery in order to compare. This study employed hematoxylin-eosin (HE) staining, echocardiography, immunofluorescence, and real-time quantitative PCR (RT-qPCR) to examine the impact of NETs on the long-term prognosis of MI. Next, datasets related to MI and NETs were downloaded from the GEO database, respectively. The Limma package of R software was used to identify differentially expressed genes (DEGs). After analyzing the \"Robust Rank Aggregation (RRA)\" package, we conducted a screening for robust differentially expressed genes (DEGs) and performed pathway enrichment analysis using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) to determine the functional roles of these robust DEGs. The protein-protein interaction (PPI) network was visualized and hub genes were filtered using Cytoscape.
RESULTS: Immunofluorescence and qPCR results showed an increase in the expression of Myeloperoxidase (MPO) at week 1 and week 8 in the hearts of mice after MI. HE staining reveals a series of pathological manifestations in the heart of the MI group during 8 weeks, including enlarged size, disordered arrangement of cardiomyocytes, infiltration of inflammatory cells, and excessive deposition of collagen fibers, among others. The utilization of Apocynin could significantly improve these poor performances. The echocardiography displayed the cardiac function of the heart in mice. The MI group has a reduced range of heart movement and decreased ejection ability. Moreover, the ventricular systolic movement was found to be abnormal, and its wall thickening rate decreased over time, indicating a progressive worsening of myocardial ischemia. The Apocynin group, on the contrary, showed fewer abnormal changes in the aforementioned aspects. A total of 81 DEGs and 4 hub genes (FOS, EGR1, PTGS2, and HIST1H4H) were obtained. The results of RT-qPCR demonstrated abnormal expression of these four genes in the MI group, which could be reversed by treatment of Apocynin.
CONCLUSIONS: The NETs formation could be highly related to MI and the long-term prognosis of MI can be significantly influenced by the NETs formation. Four hub genes, namely FOS, EGR1, PTGS2, and HIST1H4H, have the potential to be key genes related to this process. They could also serve as biomarkers for predicting MI prognosis and as targets for gene therapy.