慢加急性肝衰竭不同预后患者血浆外泌体差异蛋白的生物信息学分析

目的筛选不同预后慢加急性肝衰竭(ACLF)患者血浆外泌体中的差异蛋白,分析其功能及生物学过程,为患者临床诊断提供参考依据。方法前瞻性选取2019年7月—10月于首都医科大学附属北京佑安医院住院确诊的ACLF患者10例,随访90 d,患者死亡或肝移植归入肝移植/死亡组(n=5),患者存活归入生存组(n=5),两组一般资料指标比较采用Mann-Whitney U秩和检验。采用非标记(Label-free)定量蛋白质组学技术对血浆外泌体蛋白进行鉴定和定量分析,筛选差异蛋白并进行功能富集分析,使用R-3.5.1软件对差异蛋白进行层次聚类分析,分析其参与的生物学过程。结果外泌体蛋白质组学分析共鉴定出860种蛋白,以倍数上调>1.2倍或下调>1.2倍且P<0.05的标准筛选出差异表达蛋白116种,与肝移植/死亡组相比,生存组上调蛋白62种、下调蛋白54种。生物信息学分析结果显示,这些蛋白主要参与了免疫反应、信号转导、囊泡介导的转运、细胞死亡和增殖等生物学过程,并与炎症反应、糖类和氨基酸代谢、肝细胞损伤及再生等信号通路密切相关。结论非标记定量蛋白质组学技术筛选出的差异蛋白可能作为ACLF早期诊断及预后判断的血清学标志物。

A bioinformatics analysis of differentially expressed proteins in plasma exosome of acute-on-chronic liver failure patients with different prognoses

Objective To investigate the differentially expressed proteins in the plasma exosome of acute-on-chronic liver failure(ACLF)patients with different prognoses,to analyze their functions and biological processes,and to provide a basis for clinical diagnosis.Methods A prospective study was performed for 10 ACLF patients who were hospitalized and diagnosed in Beijing YouAn Hospital,Capital Medical University,from July 2019 to October 2019,and the patients were followed up for 90 days.The patients who died or received liver transplantation were enrolled as liver transplantation/death group(5 patients),and the patients who survived were enrolled as survival group(5 patients).The Mann-Whitney U test was used for comparison of general data between the two groups.The label-free quantitative proteomic method was used for identification and quantitative analysis of plasma exosome proteins to screen out differentially expressed proteins,and a functional enrichment analysis was performed.R-3.5.1 software was used to perform a hierarchical cluster analysis of differentially expressed proteins to analyze the biological processes involving these proteins.Results A total of 860 proteins were identified by the exosome proteomic analysis,and according to the criteria of upregulation>1.2 folds or downregulation>1.2 folds(P<0.05),there were 116 differentially expressed proteins.Compared with the liver transplantation/death group,the survival group had 62 upregulated proteins and 54 downregulated proteins.The bioinformatics analysis showed that these differentially expressed proteins mainly participated in immune reaction,signal transduction,vesicle-mediated transport,cell death,and cell proliferation and were closely associated with the signaling pathways including inflammatory response,carbohydrate and amino acid metabolism,hepatocyte injury,and hepatocyte regeneration.Conclusion Differentially expressed proteins screened out by the label-free quantitative proteomic method can be used as serological markers for the early diagnosis and prognostic evaluation of ACLF.