iTRAQ技术在颅内动脉瘤病人血浆差异表达蛋白检测中的应用

Detection of plasma differentially expressed proteins of ruptured and unruptured intracranial aneurysms using iTRAQ-based proteomics analysis

目的探讨同位素标记相对和绝对定量(iTRAQ)技术在颅内动脉瘤血浆差异表达蛋白(DEPs)检测中的应用价值。方法收集2019年1~12月收治的10例颅内破裂动脉瘤病人血浆10份,同时收集年龄、性别、吸烟史、饮酒史、高血压以及动脉瘤位置等相匹配的10例颅内未破裂动脉瘤血浆10份作为对照组。采用iTRAQ技术筛选血浆样本DEPs,应用生信分析方法GO分析和KEGG通路富集分析,并与GEO数据库的数据集进行交叉验证。结果经蛋白质组学分析鉴定出806个可定量蛋白,其中DEPs共122个,表达上调59个,表达下调63个。GO和KEGG分析结果显示,动脉瘤破裂主要与炎症和免疫反应相关。DEPs相互作用网络分析显示,白蛋白、血红蛋白β亚基、钙调蛋白1、转铁蛋白和载脂蛋白A1是网络核心蛋白。与GEO数据集交叉结果显示,血浆样本SLMAP、IQGAP3、PTPRJ、PEBP1、S100P、BASP1和AOC3等表达水平与GEO数据集的动脉瘤组织基因表达水平呈现相同变化趋势,而且破裂动脉瘤与未破裂动脉瘤之间具有统计学差异(P<0.05)。结论本文结果初步筛选出与颅内动脉瘤破裂相关的血浆DEPs,这些DEPs与炎症和免疫反应密切相关。这为研究颅内动脉瘤发病的分子机制和潜在的治疗靶点提供了新的线索。

Objective To investigate the application value of isotope labeling relative and absolute quantitative(iTRAQ)technique in detecting differential expression proteins(DEPs)in the plasma of patients with intracranial aneurysms.Methods Ten samples of plasma were collected from 10 patients with ruptured intracranial aneurysm who were admitted to our hospital from January to December 2019,and 10 samples of plasma from 10 patients with unruptured intracranial aneurysm were collected as the control group.The DEPs of plasma samples were screened by iTRAQ technique,GO analysis and KEGG pathway enrichment analysis were used to analyzed the DEPs which were cross-verified with GEO database data sets.Results A total of 806 quantifiable proteins was identified by proteomic analysis,of which 122 were DEPs(59 up-regulated,63 down-regulated).The results of GO and KEGG analyses showed that aneurysm rupture was mainly related to inflammation and immune response.DEPs interaction network analysis showed that albumin,hemoglobin subunit beta,calmodulin 1,transferrin and apolipoprotein A1 were the hub proteins of the network.The cross-over results with GEO data set showed that the expression levels of SLMAP,IQGAP3,PTPRJ,PEBP1,S100P,BASP1 and AOC3 in plasma samples presented the same change trend as those of aneurysm tissue gene expression in GEO data set.There was statistical difference between ruptured aneurysms and unruptured aneurysms(P<0.05).Conclusions The plasma DEPs associated with ruptured intracranial aneurysms were closely related to inflammation and immune response.Out results provide new clues for the study of the molecular mechanism of intracranial aneurysm pathogenesis and potential therapeutic targets.