Characterizing transcriptomic signatures and identifying hub differentially expressed genes in resistant hypertension

Resistant hypertension (RH) is one of the high-risk types within the spectrum of hypertensive disorders, characterized by a complex pathogenesis. To identify hub differentially expressed genes (DEGs) associated with this disease, this study performed transcriptome sequencing on 30 blood samples collected in 2022 from the Affiliated Hospital of Shandong University of Traditional Chinese Medicine and Jinan Fifth People's Hospital (comprising 10 hypertensive patients, 10 RH patients, and 10 healthy controls). Using DESeq2 analysis, 731 DEGs were initially screened. Subsequently, weighted gene co-expression network analysis (WGCNA) identified 2 modules significantly associated with RH (containing 1,944 genes). Taking the intersection of these module genes and the DEGs yielded 229 key DEGs. Gene Ontology (GO) enrichment analysis revealed that these key DEGs were significantly enriched in biological processes such as drug catabolic process, cellular components like hemoglobin complex, and molecular functions including peroxidase activity. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that these DEGs were associated with pathways such as the VEGF signaling pathway and mitophagy. A protein-protein interaction (PPI) network was further constructed. Using the cytohubba plugin in Cytoscape software, hub genes were identified by integrating the results from 12 algorithms (taking the intersection of the top 20 genes from each algorithm), preliminarily determining GATA1, EPB42, ANK1, and SNCA as the hub DEGs. Validation by qRT-PCR confirmed that the expression changes of GATA1 and EPB42 were consistent with the sequencing results. This study suggests that the development of RH involves the synergistic action of multiple genes, and perturbations in hub genes (GATA1, EPB42) and related pathways (VEGF signaling pathway, mitophagy) may play significant roles in the disease process. These findings provide new insights for a deeper understanding of the pathological mechanisms underlying RH.

顽固性高血压（Resistant hypertension, RH）是高血压疾病谱中的高危亚型之一，发病机制复杂。为筛选与该病相关的核心差异表达基因（differentially expressed genes, DEGs），本研究于2022年从山东中医药大学附属医院及济南市第五人民医院采集30份血液样本开展转录组测序，样本分为10名高血压患者、10名顽固性高血压患者及10名健康对照者三组。采用DESeq2进行分析后，初步筛选得到731个差异表达基因。后续通过加权基因共表达网络分析（weighted gene co-expression network analysis, WGCNA），鉴定出2个与RH显著相关的基因模块（共包含1944个基因）。将模块基因与差异表达基因取交集，最终获得229个关键差异表达基因。基因本体（Gene Ontology, GO）富集分析结果显示，这些关键差异表达基因显著富集于药物分解代谢等生物过程、血红蛋白复合物等细胞组分，以及过氧化物酶活性等分子功能类别。京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路分析表明，该类基因与血管内皮生长因子（vascular endothelial growth factor, VEGF）信号通路、线粒体自噬等通路密切相关。本研究进一步构建了蛋白质-蛋白质相互作用（protein-protein interaction, PPI）网络，借助Cytoscape软件的cytoHubba插件，整合12种算法的分析结果（取每种算法输出的前20个基因的交集）鉴定核心基因，初步确定GATA1、EPB42、ANK1及SNCA为核心差异表达基因。通过实时定量聚合酶链反应（quantitative real-time polymerase chain reaction, qRT-PCR）验证，发现GATA1与EPB42的表达变化趋势与转录组测序结果一致。本研究提示，RH的发生发展涉及多基因协同调控，核心基因（GATA1、EPB42）及相关通路（VEGF信号通路、线粒体自噬）的异常可能在疾病进程中发挥关键作用。上述研究结果为深入阐释顽固性高血压的病理机制提供了全新视角。