蛋白质组学分析小鼠动脉血栓揭示新的血栓调控通路  

Proteomics analysis of arterial thrombosis in mice reveals novel pathways of thrombosis regulation

作　　者：胡乐佳 徐鹏飞 陈虹[1] 郭子怡 宁炫 杨雪剑 饶昊杰 王淼[1] Hu Lejia;Xu Pengfei;Chen Hong;Guo Ziyi;Ning Xuan;Yang Xuejian;Rao Haojie;Wang Miao(State Key Laboratory of Cardiovascular Diseases,Fuwai Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100037,China)

机构地区：[1]北京协和医学院/中国医学科学院阜外医院国家心血管病中心,北京市100037

出　　处：《血栓与止血学》2023年第3期117-126,共10页Chinese Journal of Thrombosis and Hemostasis

基　　金：国家重点研发计划主动健康和老龄化科技应对重点专项(项目编号:2020YFC2008003);中国医学科学院医学与健康科技创新工程项目(项目编号:2021⁃I2M⁃1⁃016);2020年中国药学会⁃以岭生物医药创新基金;中国医学科学院阜外医院高水平医院临床科研业务费(项目批准号:2022⁃GSP⁃GG⁃8)。

摘　　要：目的本研究采用蛋白质组学对小鼠动脉血栓相关蛋白进行分析,明确参与调控动脉血栓形成的蛋白分子。方法应用30%氯化铁诱导小鼠颈动脉血栓形成,收集带血栓的血管组(血栓组)和颈动脉假手术血管组(非血栓组)。使用非靶向无标记液相色谱⁃质谱联用蛋白质组学定量方法对样本进行蛋白质鉴定。绘制火山图、热图并进行基因本体(GO)、京都基因与基因组百科全书(KEGG)代谢通路和蛋白质相互作用网络等生物信息学分析两组间显著差异表达蛋白(DEPs)。结果共鉴定出2896个血栓相关蛋白质,通过对血栓组与非血栓组蛋白质组学比较,火山图和热图展示了249个组间显著差异表达的蛋白(P<0.05,|Log2(fold change)|>2),其中211个表达显著上调,38个表达显著下调。GO分析和KEGG代谢通路富集分析显示,DEPs主要参与血小板活化(脱颗粒)、粘附、聚集,补体与凝血级联反应,内皮细胞受体交互作用,中性粒细胞胞外诱捕网(NETs)形成等生物学过程。其中,TGFb1、ITGa6、LY6G6F、MPIG6B、FGA、NR3C1、PDLIM2、PDGFRA与动脉血栓发生密切相关。结论除凝血系统和血小板激活之外,补体系统、天然免疫反应释放的NETs可能参与动脉血栓形成。血栓关键蛋白的发现为血栓发生机制及防治提供了新研究方向和潜在干预靶点。Objective In this study,proteomics was used to analyze the arterial thrombosis⁃related proteins in mice to clarify the protein molecules that regulate arterial thrombosis.Methods 30%ferric chloride was used to induce carotid artery thrombosis in mice.The thrombus group(thrombus group)and the carotid artery sham⁃operated group(non⁃thrombus group)were collected.The data were collected using non⁃targeted label⁃free liquid chromatography⁃mass spectrometry for protein identification of vascular samples.Volcano map and heat map were drawn,and bioinformatics analysis such as gene ontology(GO),Kyoto Encyclopedia of genes and genomes(KEGG)metabolic pathway and protein interaction network were performed to analyze significantly differentially expressed proteins(DEPs)between the two groups.Results A total of 2896 thrombosis⁃related proteins were identified.By comparing the proteomics of the thrombus group and the non⁃thrombus group,the volcano map and heat map showed 249 significantly differentially expressed proteins between the thrombus group and the non⁃thrombus group(P<0.05,|Log2(fold change)|>2),of which 211 were significantly up⁃regulated and 38 were notably down⁃regulated.GO analysis and KEGG metabolic pathway enrichment analysis showed that DEPs were mainly involved in platelet activation(degranulation),adhesion,aggregation,complement and coagulation cascade,endothelial cell receptor interaction,neutrophil extracellular traps(NETs)formation and other biological processes.Among them,TGFb1,ITGa6,LY6G6F,MPIG6B,FGA,NR3C1,PDLIM2 and PDGFRA are closely related to the regulation of arterial thrombosis.Conclusions In addition to coagulation and platelet activation,complement system and NETs released by innate immune system may also participate in arterial thrombosis.The key proteins identified here warrant further mechanistic studies and may have therapeutic implications for arterial thrombosis.

关 键 词：血栓 蛋白质组学 凝血 炎症 免疫反应 

分 类 号：R364.15[医药卫生—病理学]