Both Diabetic retinopathy (DR) and Atherosclerosis (AS) are common complications in patients with diabetes, and they share major pathophysiological similarities and have a common pathogenesis. Studies performed to date have demonstrated that ferroptosis plays a vital part in the occurrence and development of DR and AS, but its mechanism in the two diseases remains poorly understood. DR Chip data (GSE60436 and GSE102485) and AS chip data (GSE100927 and GSE57691) were obtained from the Gene Expression Omnibus (GEO) database. The screening of the differential expression genes (DEGs) was analyzed using the limma package, and the genes related to ferroptosis were obtained from the FerrDb V2 database. Two key genes (NOX4 and PARP14) were identified through external datasets validation and receiver operating characteristic (ROC) curve analysis. Gene Ontology (GO) and Gene Set Enrichment Analysis (GSEA) were used to conduct a functional enrichment analysis, and miRNA-mRNA networks were established. The CIBERSORT algorithm was applied to identify the immune cell infiltration between the disease group and control group. Next, the correlations between key genes and infiltrating immune cells were investigated by the Spearman method. Finally, the correlation between 2 key genes and ferroptosis markers was confirmed. Nine ferroptosis differentially expressed genes (DE-FRGs) between DR and AS were identified in this study. NOX4 and PARP14 were selected as key genes for further analysis by external datasets and ROC curve analysis. The key genes NOX4, PARP14 and their correlated genes (such as CYBA, NOX1, NOX3, CYBB, PARP9, PARP10, and PARP15) are mainly enriched in oxidoreductase activity, protein ADP-ribosylation, superoxide metabolic process, reactive oxygen species metabolic process, PID pathway, and VEGFA-VEGFR2 pathway. A miRNA-mRNA network was constructed, and we got 12 miRNAs correlated with the target gene NOX4, 38 miRNAs correlated with the target gene PARP14. Three common miRNAs (hsa-miR-1-3p, hsa-miR-129-2-3p, and hsa-miR-155-5p) were observed in the network. Immune infiltration analysis displayed that activated B cell, MDSC, and Type 17 T helper cell are the common immune cells involved in the immune infiltration process of DR and AS. The results revealed that there are significant correlations between two key genes and most ferroptosis marker genes no matter in DR or AS. Ferroptosis-related genes NOX4 and PARP14 may be common biomarkers of DR and AS. Both were associated with immune infiltration in patients with DR and AS. Our data provide a theoretical basis for the early diagnosis and immunotherapy of the two diseases.

中文翻译：

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两个铁死亡特异性表达基因 NOX4 和 PARP14 被认为是诊断和治疗糖尿病视网膜病变和动脉粥样硬化的潜在生物标志物

糖尿病视网膜病变（DR）和动脉粥样硬化（AS）都是糖尿病患者常见的并发症，它们具有主要的病理生理相似性和共同的发病机制。迄今为止的研究表明，铁死亡在DR和AS的发生和发展中起着至关重要的作用，但其在这两种疾病中的机制仍知之甚少。DR 芯片数据（GSE60436 和 GSE102485）和 AS 芯片数据（GSE100927 和 GSE57691）从基因表达综合（GEO）数据库获得。使用limma包对差异表达基因（DEGs）进行筛选分析，从FerrDb V2数据库中获取与铁死亡相关的基因。通过外部数据集验证和受试者工作特征（ROC）曲线分析确定了两个关键基因（NOX4 和 PARP14）。利用基因本体论（GO）和基因集富集分析（GSEA）进行功能富集分析，并建立miRNA-mRNA网络。应用CIBERSORT算法识别疾病组和对照组之间的免疫细胞浸润情况。接下来，通过Spearman方法研究关键基因与浸润免疫细胞之间的相关性。最后，证实了2个关键基因与铁死亡标记之间的相关性。本研究鉴定了 DR 和 AS 之间的 9 个铁死亡差异表达基因 (DE-FRG)。选择NOX4和PARP14作为关键基因，通过外部数据集和ROC曲线分析进行进一步分析。关键基因NOX4、PARP14及其相关基因(如CYBA、NOX1、NOX3、CYBB、PARP9、PARP10和PARP15)主要富集于氧化还原酶活性、蛋白质ADP-核糖基化、超氧化物代谢过程、活性氧代谢过程、 PID途径和VEGFA-VEGFR2途径。构建miRNA-mRNA网络，得到与靶基因NOX4相关的12个miRNA，与靶基因PARP14相关的38个miRNA。在网络中观察到三种常见的 miRNA（hsa-miR-1-3p、hsa-miR-129-2-3p 和 hsa-miR-155-5p）。免疫浸润分析显示，活化的B细胞、MDSC和17型T辅助细胞是参与DR和AS免疫浸润过程的常见免疫细胞。结果表明，无论在DR还是AS中，两个关键基因与大多数铁死亡标志基因之间都存在显着相关性。铁死亡相关基因 NOX4 和 PARP14 可能是 DR 和 AS 的常见生物标志物。两者都与 DR 和 AS 患者的免疫浸润有关。我们的数据为这两种疾病的早期诊断和免疫治疗提供了理论依据。