Yan Haijian, An Pingjiang, He Jingjing, Zhang Bin, Wei Binxing, Wu Tongqian, Li Qing
Department of Emergency, Affiliated Hospital of Guizhou Medical University, Guiyang, China.
School of Clinical Medicine, Guizhou Medical University, Guiyang, China.
Int J Genomics. 2025 Aug 11;2025:1027388. doi: 10.1155/ijog/1027388. eCollection 2025.
Spinal cord injury (SCI) is a central nervous system disorder characterized by oxidative stress and inflammatory responses. Activating transcription factor 3 (Atf3) is an early-response stress-regulating gene whose abnormal upregulation exacerbates oxidative stress and inflammation. However, despite its known association with oxidative stress, the mechanism of Atf3 in SCI remains incompletely understood. This study aimed to investigate the role and mechanisms of Atf3 in SCI. Bioinformatics analysis was performed using multiple GEO datasets to identify hub genes and key signaling pathways associated with post-SCI oxidative stress. The rat SCI model and a lipopolysaccharide (LPS)-induced PC12 cell injury model were established in vivo and in vitro to investigate whether Atf3 could ameliorate SCI progression, neuronal damage, oxidative stress, and inflammation. Chromatin immunoprecipitation quantitative PCR (ChIP-qPCR) and luciferase reporter assays were used to analyze the interaction between Atf3 and NF-B/p65, as well as the promoter activity of the p65 gene. Finally, the NF-B signaling pathway was activated to observe the role of Atf3 in neuronal injury and SCI. Bioinformatics analysis revealed significant enrichment of Atf3 and the NF-B signaling pathway in SCI datasets. Atf3 primarily colocalized with NeuN but minimally with GFAP or Iba-1. Knockdown of Atf3 significantly alleviated SCI damage, reduced oxidative stress (decreased MDA/MPO and increased SOD/GSH levels), and suppressed inflammation (reduced TNF-, IL-1 and IL-6 levels) in both SCI rats and LPS-treated PC12 cells. However, activation of NF-B counteracted these protective effects. ChIP-qPCR and luciferase reporter assays demonstrated that Atf3 overexpression enhanced its binding to the p65 promoter and promoted NF-B/p65 activation, whereas Atf3 knockdown reversed these effects. Downregulation of Atf3 mitigates oxidative stress and inflammation in SCI, potentially by regulating neuronal biological functions via the NF-B pathway. These findings provide a theoretical foundation for understanding SCI pathogenesis and identifying therapeutic targets.
脊髓损伤（SCI）是一种以氧化应激和炎症反应为特征的中枢神经系统疾病。激活转录因子3（Atf3）是一种早期应激调节基因，其异常上调会加剧氧化应激和炎症。然而，尽管已知Atf3与氧化应激有关，但其在脊髓损伤中的作用机制仍不完全清楚。本研究旨在探讨Atf3在脊髓损伤中的作用及机制。使用多个GEO数据集进行生物信息学分析，以识别与脊髓损伤后氧化应激相关的枢纽基因和关键信号通路。在体内和体外建立大鼠脊髓损伤模型和脂多糖（LPS）诱导的PC12细胞损伤模型，以研究Atf3是否可以改善脊髓损伤的进展、神经元损伤、氧化应激和炎症。采用染色质免疫沉淀定量PCR（ChIP-qPCR）和荧光素酶报告基因检测分析Atf3与NF-κB/p65之间的相互作用，以及p65基因的启动子活性。最后，激活NF-κB信号通路，观察Atf3在神经元损伤和脊髓损伤中的作用。生物信息学分析显示，Atf3和NF-κB信号通路在脊髓损伤数据集中显著富集。Atf3主要与NeuN共定位，但与GFAP或Iba-1的共定位最少。敲低Atf3可显著减轻脊髓损伤大鼠和LPS处理的PC12细胞中的脊髓损伤，降低氧化应激（降低MDA/MPO水平并提高SOD/GSH水平），并抑制炎症（降低TNF-α、IL-1和IL-6水平）。然而，NF-κB的激活抵消了这些保护作用。ChIP-qPCR和荧光素酶报告基因检测表明，Atf3过表达增强了其与p65启动子的结合并促进NF-κB/p65激活，而Atf3敲低则逆转了这些作用。Atf3的下调减轻了脊髓损伤中的氧化应激和炎症，可能是通过NF-κB途径调节神经元生物学功能实现的。这些发现为理解脊髓损伤发病机制和确定治疗靶点提供了理论基础。
