Shao Qiaoyu, Wang Hao, Li Shicheng, Zeng Mengying, Zhang Shuyang, Yan Xiaowei
Department of Cardiology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China.
Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Int J Mol Sci. 2025 Apr 15;26(8):3722. doi: 10.3390/ijms26083722.
Salt-sensitive hypertension (SSH) is closely associated with arterial inflammation, yet its molecular mechanisms remain unclear. In this study, we utilized deoxycorticosterone acetate (DOCA)-salt-induced hypertensive mice, which exhibited elevated blood pressure and significant arterial inflammation. Single-cell RNA sequencing (scRNA-seq) identified interferon regulatory factor 5 (IRF5) and its downstream targets, signal transducer and activator of transcription (STAT), as key regulators of these inflammatory changes. In vivo, IRF5 levels were significantly elevated in the DOCA group, while STAT1 and STAT2 protein levels were comparable to those in the normal salt group. However, nuclear levels of phosphorylated STAT1 (pSTAT1) and phosphorylated STAT2 (pSTAT2) were markedly higher in the DOCA group. Furthermore, scRNA-seq analysis showed increased IRF5 expression in endothelial cells (ECs) in both human and mouse aorta samples. In vitro, IRF5 knockdown in artery ECs led to a reduction in nuclear pSTAT1 and pSTAT2 expression. These results suggest that IRF5 promotes STAT1 and STAT2 phosphorylation, enabling their nuclear translocation. Additionally, RNA sequencing indicated a positive correlation between endothelial cell-specific molecule 1 (ESM1) and STAT1/STAT2. Using the UCSC and JASPAR databases, we identified multiple binding sites for the STAT1::STAT2 dimer on the ESM1 promoter. Luciferase reporter assays revealed enhanced ESM1 transcription following pSTAT1::pSTAT2 binding, and pinpoint potential binding sites. Chromatin Immunoprecipitation Quantitative PCR (ChIP-qPCR) further confirmed the specific binding sites between the pSTAT1::pSTAT2 dimer and the ESM1 promoter. These findings highlight the critical role of the IRF5-pSTAT1::pSTAT2-ESM1 pathway in the pathogenesis of SSH and suggest potential therapeutic targets.
盐敏感性高血压（SSH）与动脉炎症密切相关，但其分子机制仍不清楚。在本研究中，我们利用醋酸脱氧皮质酮（DOCA）-盐诱导的高血压小鼠，这些小鼠表现出血压升高和显著的动脉炎症。单细胞RNA测序（scRNA-seq）确定干扰素调节因子5（IRF5）及其下游靶点信号转导和转录激活因子（STAT）是这些炎症变化的关键调节因子。在体内，DOCA组中IRF5水平显著升高，而STAT1和STAT2蛋白水平与正常盐组相当。然而，DOCA组中磷酸化STAT1（pSTAT1）和磷酸化STAT2（pSTAT2）的核水平明显更高。此外，scRNA-seq分析显示，在人和小鼠主动脉样本的内皮细胞（ECs）中IRF5表达增加。在体外，动脉内皮细胞中IRF5的敲低导致核pSTAT1和pSTAT2表达减少。这些结果表明，IRF5促进STAT1和STAT2磷酸化，使其能够进行核转位。此外，RNA测序表明内皮细胞特异性分子1（ESM1）与STAT1/STAT2之间呈正相关。利用UCSC和JASPAR数据库，我们在ESM1启动子上鉴定了多个STAT1::STAT2二聚体的结合位点。荧光素酶报告基因检测显示，pSTAT1::pSTAT2结合后ESM1转录增强，并确定了潜在的结合位点。染色质免疫沉淀定量PCR（ChIP-qPCR）进一步证实了pSTAT1::pSTAT2二聚体与ESM1启动子之间的特异性结合位点。这些发现突出了IRF5-pSTAT1::pSTAT2-ESM1通路在SSH发病机制中的关键作用，并提示了潜在的治疗靶点。
