基于蛋白组学的血友病性关节炎差异蛋白生物信息学比较研究

Comparative Bioinformatics Study of Differential Proteins in Hemophilic Arthropathy Based on Proteomics

摘要: 目的：探讨血友病性关节病(HA)滑膜组织的蛋白质表达特征，并与骨关节炎(OA)和类风湿关节炎(RA)滑膜组织进行比较，以筛选HA相关差异表达蛋白及潜在分子通路。方法：收集接受全膝关节置换术患者的HA、OA和RA滑膜组织样本，每组各7例。采用数据独立采集(DIA)定量蛋白质组学技术进行蛋白质检测，并通过MaxQuant、Andromeda及MSstats等工具完成蛋白鉴定、定量及差异表达分析。进一步对差异表达蛋白进行GO功能注释、KEGG通路富集分析及蛋白质互作网络(PPI)分析。结果：本研究共定量鉴定出80,931条肽段和6151种蛋白质。Venn分析显示，HA与OA及HA与RA比较中共有415种共同差异表达蛋白。OTUD6B表现出显著差异表达，MMP3在HA组织中呈低表达趋势。GO分析显示，差异蛋白主要涉及蛋白质运输、mRNA加工、RNA剪接、信号转导、金属离子结合及线粒体相关结构。KEGG和PPI分析提示，PI3K-Akt信号通路、氧化磷酸化通路和代谢通路显著富集，其中氧化磷酸化通路在PPI网络中具有密集互作关系。结论：HA滑膜组织具有区别于OA和RA的蛋白表达特征，其发生发展可能与铁沉积、氧化应激、线粒体能量代谢异常、蛋白稳态失衡及细胞信号转导改变有关。

Abstract: Objective: To investigate the proteomic characteristics of synovial tissues from patients with hemophilic arthropathy (HA) and compare them with those from patients with osteoarthritis (OA) and rheumatoid arthritis (RA), aiming to identify HA-related differentially expressed proteins and potential molecular pathways. Methods: Synovial tissue samples were collected from patients with end-stage HA, OA and RA who underwent total knee arthroplasty, with seven samples included in each group. Data-independent acquisition (DIA)-based quantitative proteomics was performed to characterize protein expression profiles. Protein identification, quantification and differential expression analysis were conducted using MaxQuant, the Andromeda search engine and MSstats. Gene Ontology (GO) annotation, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and protein-protein interaction (PPI) network analysis were further performed to explore the biological functions of differentially expressed proteins. Results: A total of 80,931 peptides and 6151 proteins were quantitatively identified. Venn analysis identified 415 common differentially expressed proteins in HA compared with both OA and RA. OTUD6B showed marked differential expression, while MMP3 displayed a relatively low expression trend in HA synovial tissues. GO analysis indicated that the differentially expressed proteins were mainly associated with protein transport, mRNA processing, RNA splicing, signal transduction, metal ion binding and mitochondrial components. KEGG and PPI analyses revealed significant enrichment of the PI3K-Akt signaling pathway, oxidative phosphorylation and metabolic pathways, with oxidative phosphorylation showing extensive interactions in the PPI network. Conclusion: HA synovial tissues exhibit distinct proteomic features compared with OA and RA tissues. The pathogenesis of HA may be associated with iron deposition, oxidative stress, mitochondrial energy metabolism dysfunction, impaired protein homeostasis and altered cellular signaling.