螨虫诱导变应性鼻炎患者血浆细胞外囊泡的蛋白质组学研究

Proteomic study of plasma extracellular vesicles in house dust mite-induced allergic rhinitis patients

目的探究螨虫诱导的变应性鼻炎(allergic rhinitis,AR)患者血浆细胞外囊泡(extracellular vesicle,EV)蛋白质组学的差异,寻找并鉴定AR潜在的生物标志物。方法分别收集螨虫诱导的AR患者(AR组)和健康志愿者(HC组)的血液样本,每组各20例。使用超速离心法对AR组与HC组的血浆EV进行分离,并使用透射电子显微镜和纳米流式细胞术对EV进行鉴定和表征。使用液相色谱-串联质谱技术对分离出的EV进行蛋白质组学分析,寻找并鉴定差异表达蛋白质(differentially expression proteins,DEPs)。随后,对DEPs进行基因本体(gene ontology,GO)、京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析和蛋白质相互作用(protein-protein interaction,PPI)网络分析。结果AR组和HC组的血浆EV大小在30~150 nm之间,两组之间无显著性差异。在两组之间共筛选出664种蛋白质,并鉴定出89种表达水平具有显著性差异的蛋白质(P<0.05)。与HC组相比,AR组中有44种蛋白质表达显著上调,45种蛋白质表达显著下调。通过进行GO分析,发现这89种DEPs可能与免疫调节有关。此外,KEGG富集分析和PPI分析显示,这89种DEPs还与补体和凝血级联反应、雌激素和脂肪细胞因子信号通路等多种途径密切相关。结论本研究通过蛋白质组学技术成功筛选出与螨虫诱导AR相关的89种差异表达蛋白质。这些DEPs主要参与调节个体的免疫反应,其中脂联素、补体C1q亚基A和溶质载体家族2等蛋白质可能成为诊断AR的潜在生物标志物。

Objective To explore the differences in the proteomic profiles of extracellular vesicles(EVs)in the plasma of patients with house dust mite-induced allergic rhinitis(AR)and identify potential biomarkers for AR.Methods Blood samples were collected from 20 patients with house dust mite-induced AR(AR group)and 20 healthy individuals(HC group).Plasma EVs were isolated from the AR and HC groups using ultracentrifugation,and EVs were characterized and identified using transmission electron microscopy and nanoflow cytometry.Proteomic analysis of the isolated EVs was performed using liquid chromatography-mass spectrometry to identify differentially expressed proteins(DEPs).Gene ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis,and protein-protein interaction(PPI)network analysis were conducted for the DEPs.Results The size of plasma EVs from both the AR and HC groups ranged from 30 to 150 nm,with no significant differences between the two groups.A total of 664 proteins were screened and 89 proteins were identified as DEPs with significant expression differences(P<0.05)between the AR and HC groups.Compared to the HC group,the AR group showed upregulation of 44 proteins and downregulation of 45 proteins.GO analysis revealed that these 89 DEPs may be associated with immune regulation.Furthermore,KEGG pathway enrichment analysis and PPI analysis demonstrated that these DEPs were closely related to complement and coagulation cascades,estrogen signaling pathway,and adipokine signaling pathway,among other pathways.Conclusions This study successfully identified that 89 DEPs associated with house dust mite-induced AR through proteomic analysis of EVs.These DEPs are primarily involved in regulating immune responses.Proteins such as adiponectin,complement C1q subcomponent subunit A,and solute carrier family 2,facilitated glucose transporter member 1 may serve as potential biomarkers for diagnosing AR.