Xu Min, Wu Yinxiang, Meng Jiaqi, Chen Mengchen, Ding Chen
Department of Trauma Orthopedic, First Affiliated Hospital of Naval Medical University, Shanghai, China.
Department of Pulmonary and Critical Care Medicine, Third Affiliated Hospital of Naval Medical University, Shanghai, China.
Front Endocrinol (Lausanne). 2025 Jul 7;16:1606877. doi: 10.3389/fendo.2025.1606877. eCollection 2025.
This study aims to elucidate the molecular mechanisms by which the widely used organophosphate flame retardant and plasticizer, triphenyl phosphate (TPhP), disrupts bone metabolism, highlighting the potential impact of environmental chemicals on bone homeostasis.
A combined approach of network toxicology and molecular docking was employed to investigate the molecular mechanisms underlying the effects of TPhP on bone metabolism. Potential targets associated with both TPhP and bone metabolism were identified through database searches in ChEMBL, STITCH, GeneCards, and OMIM. A protein-protein interaction (PPI) network was constructed using the STRING database and analyzed with Cytoscape software. Functional enrichment analyses, including Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, were performed to determine the major pathways involved. Molecular docking was conducted to evaluate the binding affinity between TPhP and key target proteins. Additionally, experiments using MC3T3-E1 osteoblasts were conducted to validate the bioinformatics findings.
78 potential targets related to both TPhP and bone metabolism were identified. STRING and Cytoscape revealed six key proteins: IGF1R, NR3C1, MAP3K1, BRAF, WNK4, and CNR2. GO and KEGG analyses indicated that these targets predominantly function through the MAPK signaling pathway. Molecular docking results demonstrated strong binding affinities between TPhP and key targets, particularly BRAF and WNK4. , TPhP inhibited osteoblast proliferation and migration in a dose-dependent manner and downregulated EMT-related proteins and key target genes via MAPK signaling.
TPhP disrupts bone metabolism by modulating key proteins and pathways, underscoring its potential health risks and the need for further epidemiological and clinical research.
本研究旨在阐明广泛使用的有机磷酸酯阻燃剂和增塑剂磷酸三苯酯（TPhP）破坏骨代谢的分子机制，突出环境化学物质对骨稳态的潜在影响。
采用网络毒理学和分子对接相结合的方法，研究TPhP对骨代谢影响的分子机制。通过在ChEMBL、STITCH、GeneCards和OMIM数据库中搜索，确定与TPhP和骨代谢相关的潜在靶点。使用STRING数据库构建蛋白质-蛋白质相互作用（PPI）网络，并用Cytoscape软件进行分析。进行功能富集分析，包括基因本体（GO）注释和京都基因与基因组百科全书（KEGG）通路分析，以确定主要涉及的通路。进行分子对接以评估TPhP与关键靶蛋白之间的结合亲和力。此外，使用MC3T3-E1成骨细胞进行实验以验证生物信息学结果。
确定了78个与TPhP和骨代谢相关的潜在靶点。STRING和Cytoscape揭示了6个关键蛋白：IGF1R、NR3C1、MAP3K1、BRAF、WNK4和CNR2。GO和KEGG分析表明，这些靶点主要通过MAPK信号通路发挥作用。分子对接结果表明TPhP与关键靶点之间具有很强的结合亲和力，尤其是BRAF和WNK4。TPhP以剂量依赖性方式抑制成骨细胞增殖和迁移，并通过MAPK信号下调EMT相关蛋白和关键靶基因。
TPhP通过调节关键蛋白和通路破坏骨代谢，强调了其潜在的健康风险以及进一步进行流行病学和临床研究的必要性。
