摘要
目的探究钙卫蛋白S100A8/A9对肝星状细胞(HSC)的直接刺激效应及其潜在调控机制。方法通过不同浓度(200、1000 ng/mL)S100A8/A9异源二聚体重组蛋白刺激小鼠原代HSC,利用基于质谱的定量蛋白质组学串联质量标签(TMT)标记技术获得定量蛋白质组学数据,分析HSC经S100A8/A9重组蛋白刺激后蛋白表达谱的变化。采用Significance B分析方法,依据P<0.05筛选差异蛋白并进行Reactome通路富集分析,进一步基于STRING数据库对通路中的差异蛋白进行蛋白-蛋白相互作用分析。结果高浓度S100A8/A9(1000 ng/mL)处理显著改变了HSC蛋白表达谱,Reactome通路富集分析发现,转化生长因子-β(TGF-β)信号转导、核因子κB(NF-κB)信号激活、细胞因子介导的免疫调控,以及胶原生物合成等通路显著富集,蛋白-蛋白互作分析发现NF-κB转录因子亚基(RELB)、C-X-C基序趋化因子配体10(CXCL10)和Notch受体1(NOTCH1)是调控网络中的关键节点蛋白。结论高浓度S100A8/A9可直接促进HSC活化,其机制可能与NF-κB、TGF-β和Notch信号通路的调控有关,为S100A8/A9对HSC直接激活效应的机制研究提供重要参考。
Objective To investigate the direct stimulatory effects of calprotectin S100A8/A9 on hepatic stellate cells(HSCs)and underlying regulatory mechanisms.Methods Primary HSCs of mice were stimulated with S100A8/A9 heterodimer recombinant protein at 200 and 1000 ng/mL.Data on quantitative proteomics was obtained using the tandem mass tag(TMT)-labeled method before changes in the protein level of HSCs were analyzed.Differentially expressed proteins(DEPs)were screened using Significance B method and P<0.05,followed by Reactome pathway enrichment analysis.Furthermore,protein-protein interactions between the DEPs enriched in the pathways were analyzed using the STRING database.Results The protein expression profile of HSCs was significantly altered after treatment with S100A8/A9 at 1000 ng/mL.Reactome pathway enrichment analysis revealed significant enrichment in such pathways as transforming growth factor-beta(TGF-β)signaling,nuclear factor kappa-B(NF-κB)signaling activation,cytokine-mediated immune regulation,and collagen biosynthesis.The analysis of protein-protein interactions identified NF-kappa-B transcription factor subunit(RELB),chemokine(C-X-C motif)ligand 10(CXCL10)and Notch receptor 1(NOTCH1)as key hub proteins in the regulatory network.Conclusion S100A8/A9 can directly stimulate the activation of HSCs,through NF-κB signaling,TGF-βsignaling,and Notch signaling pathways potentially.This study sheds light on the mechanisms underlying the activation of HSCs stimulated by S100A8/A9.
作者
刘睿茜
刘金芳
王建
徐平
LIU Ruixi;LIU Jinfang;WANG Jian;XU Ping(State Key Laboratory of Medical Proteomics,Beijing Proteome Research Center,National Center for Protein Sciences(Beijing),Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 102206,China;Taikang Medical School(School of Basic Medical Sciences),Wuhan University,Wuhan 430071,China;Research Unit of Proteomics&Research and Development of New Drug,Chinese Academy of Medical Sciences,Beijing 102206,China)
出处
《军事医学》
2025年第10期721-727,共7页
Military Medical Sciences
基金
国家自然科学基金项目(32141003,32371503)
国家重点研发计划(2024YFC3405400)
北京市自然科学基金-大兴创新联合基金(L246037)
医学蛋白质组学全国重点实验室项目(SKLP-K202201)。
