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Quantitative Proteomic Analysis of Peripheral Blood Mononuclear Cells in Ankylosing Spondylitis by iTRAQ

机译:用iTRAQ定量分析强直性脊柱炎患者外周血单个核细胞的蛋白质组学

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摘要

This study was designed to identify and quantify the different proteins expression levels in ankylosing spondylitis (AS) and to explore the pathogenesis of AS. We performed isobaric tags for relative and absolute quantitation (iTRAQ) coupled with multiple chromatographic fractionation and tandem mass spectrometry to detect the proteins profiling in peripheral blood mononuclear cells (PBMCs) from AS patients and healthy controls. Mascot software and the International Protein Index and the Gene Ontology (GO) database were used to conduct the bioinformatics analysis. The differentially expressed proteins were validated by enzyme-linked immunosorbent assay (ELISA). A total of 1,232 proteins were identified by iTRAQ, of which 183 showed differential expression and 18 differentially expressed proteins were acute phase reactants. Upon mapping of the differentially expressed proteins to GO database, we found four differentially expressed proteins involved in the biological process of cell killing, including up-regulated cathepsin G (CTSG), neutrophil defensin3 (DEFA3), protein tyrosine phosphatase receptor type C (PTPRC), and down-regulated peroxiredoxin-1(PRDX1),which were consistent with the verified results of ELISA. Our proteomic analyses suggested that the proteins involved in the biological process of cell killing might play an important role in the pathogenesis of AS.
机译:本研究旨在鉴定和定量强直性脊柱炎(AS)中不同蛋白质的表达水平,并探讨AS的发病机理。我们进行了相对定量和绝对定量(iTRAQ)的等压标记,并进行了多次色谱分离和串联质谱分析,以检测AS患者和健康对照组的外周血单个核细胞(PBMC)中的蛋白质谱。使用Mascot软件以及国际蛋白质索引和基因本体论(GO)数据库进行生物信息学分析。差异表达的蛋白质通过酶联免疫吸附测定(ELISA)进行了验证。 iTRAQ共鉴定了1,232种蛋白质,其中183种显示差异表达,而18种差异表达蛋白是急性期反应物。通过将差异表达的蛋白质映射到GO数据库后,我们发现了四种差异表达的蛋白质参与了细胞杀伤的生物学过程,包括上调的组织蛋白酶G(CTSG),中性粒细胞防御素3(DEFA3),蛋白酪氨酸磷酸酶受体C(PTPRC) ),并下调peroxiredoxin-1(PRDX1),与ELISA验证结果一致。我们的蛋白质组学分析表明,参与细胞杀伤生物学过程的蛋白质可能在AS的发病机理中起重要作用。

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