布鲁氏菌病患者尿液差异蛋白质组学分析

Differential proteomic analysis of urine in patients with brucellosis

目的通过对布鲁氏菌病(简称布病)患者尿液蛋白进行通量化鉴定,发现与诊断布病相关的候选标示蛋白,并挖掘其参与的信号通路和生物标志物应用信息。方法收集布病患者及非布病患者尿液样本各10例,采用基于质谱的非标定量蛋白质组学技术,发现与布病相关的差异表达蛋白,通过高通量生物信息学软件深入挖掘,确定差异表达蛋白参与的疾病相关通路并筛选生物标志分子。结果本研究定性鉴定到1 817个蛋白,以|Fold Change (log2)|>1.5和BH法校正P<0.05为标准,发现尿液中布病相关差异表达蛋白185个(13个蛋白表达上调,172个蛋白表达下调),其中有17个蛋白在人的尿液中首次发现并呈现疾病表达差异;布病尿液样本中胰岛素样生长因子2(IGF2)缺失表达,其与蛋白CD300分子样家族成员f(CD300LF)、血清类粘蛋白1和2(ORM1,ORM2)、富含亮氨酸α-2-糖蛋白1(LRG1)、SH3结构域谷氨酸富集样蛋白3(SH3BGRL3)、半胱氨酸丰富跨膜BMP调节剂1(CRIM1)和谷氨酸氨连接酶(GLUL)是潜在的诊断布病、预测布病并发症的尿液标示蛋白。结论通过差异蛋白质组学分析布病患者尿液中差异表达的蛋白,为布病的快速诊断、发现药物靶点提供了新的生物标志分子,同时也为布病发病机制的揭示提供了分子线索。

To identify potential urine marker proteins for the diagnosis of brucellosis,the urine samples of 10 patients with brucellosis and 10 people without brucellosis were identified by UPLC-MS/MS.On the basis of label-free quantitative proteomics,the raw data were searched in Proteome Discoverer software,and IPA software was used to analyze canonical pathways and biomarker applications of differentially expressed prote ins.Finally,1817 proteins were identified,among which 185 differentially expressed proteins(13 up-regulated and 172 down-regulated)were screened according to|fold change(log 2)|>1.5 and BH correction P<0.05.Seventeen previously unreported proteins were detected in human urine and showed differential expression in brucellosis.IGF2 was not found in the urine samples of patients with brucellosis.IGF2,CD300LF,ORM1,ORM2,LRG1,SH3BGRL3,CRIM1 and GLUL are potential urine marker proteins for diagnosing brucellosis and predicting its complications.This study not only provides new biomarkers for the rapid diagnosis of brucellosis and the discovery of drug targets,but also provides molecular clues for revealing the pathogenesis of this disease.