基于蛋白质组学的支气管哮喘(巴达干赫依偏盛型)血清差异蛋白变化的机制研究

Proteomics-based screening of differentially expressed protein in bronchial asthma(syndrome of excessive cold)

采用蛋白质组学寻找可能与支气管哮喘(bronchial asthma, BA)致病相关的关键蛋白。首先收集健康成年人与哮喘患者的血清样本,运用去除高丰度结构和非特异性蛋白质后的蛋白定量质谱(Tandem Mass TagTM,TMT),分别获得哮喘患者与健康成年人差异表达蛋白,并进行GO富集和KEGG富集分析,通过蛋白相互作用(PPI)网络分析筛选出哮喘组中的核心蛋白。然后将核心蛋白在3例支气管哮喘患者和3例健康成年人中进行蛋白印迹(Western blot)验证。通过蛋白质组学筛查共鉴定出778个差异表达蛋白,其中32个蛋白质包含定量信息(18个蛋白表达上调、14个蛋白表达下调);KEGG富集分析具有明显表达差异的信号通路有28条。PPI结果显示有10个蛋白(GDN、1433Z、VWF、HEMO、CERU、A1AT、TSP1、G3P、IBP7、KPYM)可能在支气管哮喘的发病机制中扮演重要作用。而相对于健康成年人,支气管哮喘患者血清中SLC25A4、SVEP1和KRT25蛋白表达水平升高,差异均具有统计学意义(P<0.05)。因此,推测多种免疫信号通路及差异表达蛋白在BA的发病机制中扮演了重要作用,为BA的治疗靶点研究进一步提供了方向。

Proteomic tools were used to identify the key proteins that might be associated with bronchial asthma(BA). Firstly, the serum samples from healthy adults and asthmatic patients were collected. Tandem Mass TagTM(TMT), which removes high-abundance structures and nonspecific proteins, was employed to identify the differentially expressed proteins between asthmatic patients and healthy adults. Gene Ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were carried out for the differentially expressed proteins. The core proteins in the asthma group were screened out by protein-protein interaction(PPI) analysis. Then the core proteins were verified by Western blot for 3 patients with bronchial asthma and 3 healthy adults. A total of 778 differentially expressed proteins were screened out, among which 32 proteins contained quantitative information, including 18 up-regulated proteins and 14 down-regulated proteins. The differentially expressed proteins were enriched in 28 KEGG signaling pathways. The PPI analysis showed that 10 proteins(GDN, 1433 Z, VWF, HEMO, CERU, A1 AT, TSP1, G3 P, IBP7, and KPYM) might be involved in the pathogenesis of bronchial asthma. Compared with those in healthy adults, the expression levels of SLC25 A4, SVEP1, and KRT25 in the sera of asthmatic patients were up-regulated(P<0.05). Therefore, it is hypothesized that a variety of immune signaling pathways and differentially expressed proteins play a role in the pathogenesis of BA, which provides potential target information for the treatment of BA.