Li Pengpeng, Tao Zhenxin, Gao Yangyang, Mu Zhengqian, Tian Jiajia, Zhang YaTing, Yang Wenhui, Li Yilu, Zhao Xudong
Wuxi School of Medicine, Jiangnan University, Wuxi, Jiangsu, China.
Department of Neurosurgery, Jiangnan University Medical Center, Wuxi, Jiangsu, China.
Brain Behav. 2025 May;15(5):e70493. doi: 10.1002/brb3.70493.
This study aimed to investigate the role of secretory phosphoprotein 1 (SPP1/OPN) in modulating iron-induced cell death (ferroptosis) following intracerebral hemorrhage (ICH). By integrating transcriptomic analysis and experimental validation, we sought to identify key molecular pathways and therapeutic targets associated with ferroptosis in ICH.
The Gene Expression Omnibus Series GSE24265 dataset was analyzed using the limma package (R platform) to identify differentially expressed genes. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) enrichment analyses were performed to elucidate biological functions. Genes associated with iron-induced mortality were identified by cross-referencing transcriptomic profiles with the FerrDb database. A protein-protein interaction network was constructed using Cytoscape, and hub genes were identified. An experimental ICH model was developed in mice using stereotactic instrumentation, and the effects of OPN administration were evaluated through neurological assessments, biochemical assays (superoxide dismutase, glutathione, malondialdehyde), Western immunoblotting (GPX4, ACSL4), Prussian blue histochemistry, and electron microscopy.
Transcriptomic analysis identified 27 hub genes, with CD44 and ITGB3 characterized as receptors for OPN. In the ICH model, OPN administration improved neurological outcomes, elevated antioxidant markers, and reduced lipid peroxidation. OPN upregulated GPX4 while suppressing ACSL4, indicating anti-ferroptotic effects. These protective effects were mediated through the Nrf2 pathway, as confirmed by inhibitor ML385. Prussian blue staining and electron microscopy demonstrated reduced cerebral iron deposition and mitochondrial damage following OPN treatment.
This study provides novel evidence for SPP1/OPN as a key modulator of ferroptosis in ICH, highlighting its potential as a therapeutic target. By enhancing iron homeostasis and mitigating oxidative stress, OPN offers a promising strategy for improving outcomes in ICH patients.
本研究旨在探讨分泌型磷蛋白1（SPP1/骨桥蛋白，OPN）在脑出血（ICH）后调节铁诱导的细胞死亡（铁死亡）中的作用。通过整合转录组分析和实验验证，我们试图确定与ICH中铁死亡相关的关键分子途径和治疗靶点。
使用limma软件包（R平台）分析基因表达综合数据库GSE24265数据集，以鉴定差异表达基因。进行京都基因与基因组百科全书（KEGG）通路和基因本体论（GO）富集分析，以阐明生物学功能。通过将转录组谱与FerrDb数据库交叉参照，鉴定与铁诱导死亡相关的基因。使用Cytoscape构建蛋白质-蛋白质相互作用网络，并鉴定枢纽基因。使用立体定位仪器在小鼠中建立实验性ICH模型，并通过神经学评估、生化测定（超氧化物歧化酶、谷胱甘肽、丙二醛）、Western免疫印迹（GPX4、ACSL4）、普鲁士蓝组织化学和电子显微镜评估OPN给药的效果。
转录组分析鉴定出27个枢纽基因，其中CD44和ITGB3被表征为OPN的受体。在ICH模型中，给予OPN可改善神经学结果，提高抗氧化标志物水平，并减少脂质过氧化。OPN上调GPX4，同时抑制ACSL4，表明具有抗铁死亡作用。如抑制剂ML385所证实，这些保护作用是通过Nrf2途径介导的。普鲁士蓝染色和电子显微镜显示，OPN治疗后脑铁沉积和线粒体损伤减少。
本研究为SPP1/OPN作为ICH中铁死亡的关键调节因子提供了新证据，突出了其作为治疗靶点的潜力。通过增强铁稳态和减轻氧化应激，OPN为改善ICH患者的预后提供了一种有前景的策略。
