运用尿蛋白组学技术探索凝血酶与炎症因子相互作用参与IgA血管炎发病的机制

刘萌萌; 侯改灵; 杨晓青; 张秋爽; 梅晓峰; 丁樱; 宋兰; 黄岩杰

doi:10.7499/j.issn.1008-8830.2311151

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中国当代儿科杂志 ›› 2024, Vol. 26 ›› Issue (7) : 683-689. DOI: 10.7499/j.issn.1008-8830.2311151

论著·临床研究

运用尿蛋白组学技术探索凝血酶与炎症因子相互作用参与IgA血管炎发病的机制

刘萌萌^1,2, 侯改灵^1,2, 杨晓青², 张秋爽², 梅晓峰², 丁樱^1,2, 宋兰³, 黄岩杰^1,4

作者信息 +

Exploring the mechanism of IgA vasculitis pathogenesis through the interaction of thrombin and inflammatory factors using urinary proteomics

LIU Meng-Meng, HOU Gai-Ling, YANG Xiao-Qing, ZHANG Qiu-Shuang, MEI Xiao-Feng, DING Ying, SONG Lan, HUANG Yan-Jie

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文章历史 +

摘要

目的运用蛋白质组学方法探索高凝状态参与IgA血管炎（IgA vasculitis, IgAV）发病的证据、尿生物标志物及部分机制。方法采用高效液相色谱-串联质谱技术筛选10例正常儿童和10例IgAV患儿尿液中差异表达的蛋白，进行Reactome通路分析。运用STRING 和Cytoscape软件进行蛋白质-蛋白质相互作用（protein-protein interaction, PPI）网络分析。在验证队列中，纳入15例正常儿童和25例IgAV患儿，采用酶联免疫吸附测定方法验证尿液差异蛋白的表达水平。结果 IgAV患儿与正常组之间共筛选出772个差异蛋白（上调蛋白768个，下调蛋白4个）。Reactome通路富集结果显示中性粒细胞脱颗粒、血小板活化及止血通路参与IgAV的发生。差异蛋白中巨噬细胞迁移抑制因子（macrophage migration inhibitory factor, MIF）在中性粒细胞脱颗粒和止血过程中起着重要作用，凝血酶是参与血小板活化和止血通路的关键蛋白。PPI分析结果显示，凝血酶与多种参与炎症反应的差异蛋白具有直接作用，并通过它们与MIF相互作用。验证结果显示，与正常儿童相比，IgAV患儿尿凝血酶/肌酐、尿MIF/肌酐水平均显著升高（P<0.05）。结论凝血酶通过与炎症因子相互作用参与IgAV的发病；尿凝血酶和MIF可作为反映IgAV患儿高凝与炎症状态的生物标志物。

Abstract

Objective To explore the evidence, urinary biomarkers, and partial mechanisms of hypercoagulability in the pathogenesis of IgA vasculitis (IgAV). Methods Differential expression of proteins in the urine of 10 healthy children and 10 children with IgAV was screened using high-performance liquid chromatography-tandem mass spectrometry, followed by Reactome pathway analysis. Protein-protein interaction (PPI) network analysis was conducted using STRING and Cytoscape software. In the validation cohort, 15 healthy children and 25 children with IgAV were included, and the expression levels of differential urinary proteins were verified using enzyme-linked immunosorbent assay. Results A total of 772 differential proteins were identified between the IgAV group and the control group, with 768 upregulated and 4 downregulated. Reactome pathway enrichment results showed that neutrophil degranulation, platelet activation, and hemostasis pathways were involved in the pathogenesis of IgAV. Among the differential proteins, macrophage migration inhibitory factor (MIF) played a significant role in neutrophil degranulation and hemostasis, while thrombin was a key protein in platelet activation and hemostasis pathways. PPI analysis indicated that thrombin directly interacted with several proteins involved in inflammatory responses, and these interactions involved MIF. Validation results showed that compared to healthy children, children with IgAV had significantly higher urine thrombin/creatinine and urine MIF/creatinine levels (P<0.05). Conclusions Thrombin contributes to the pathogenesis of IgAV through interactions with inflammatory factors. Urinary thrombin and MIF can serve as biomarkers reflecting the hypercoagulable and inflammatory states in children with IgAV.

导出引用

刘萌萌, 侯改灵, 杨晓青, 张秋爽, 梅晓峰, 丁樱, 宋兰, 黄岩杰. 运用尿蛋白组学技术探索凝血酶与炎症因子相互作用参与IgA血管炎发病的机制[J]. 中国当代儿科杂志. 2024, 26(7): 683-689 https://doi.org/10.7499/j.issn.1008-8830.2311151

LIU Meng-Meng, HOU Gai-Ling, YANG Xiao-Qing, ZHANG Qiu-Shuang, MEI Xiao-Feng, DING Ying, SONG Lan, HUANG Yan-Jie. Exploring the mechanism of IgA vasculitis pathogenesis through the interaction of thrombin and inflammatory factors using urinary proteomics[J]. Chinese Journal of Contemporary Pediatrics. 2024, 26(7): 683-689 https://doi.org/10.7499/j.issn.1008-8830.2311151

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基金

国家自然科学基金面上项目（82174187）；中原科技创新领军人才项目（234200510028）；河南省中医药学科领军人才项目（［2021］No. 8）；河南省卫生健康委国家中医临床研究基地科研专项（2022JDZX014）；河南省特色骨干学科中医学学科建设项目（STG-ZYXKY-2020008）。