Identification of Potential Serum Biomarkers for Rheumatoid Arthritis by High-Resolution Quantitative Proteomic Analysis

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• 作者：Yongjing Cheng ; Yuling Chen ; Xiaolin Sun ; Yuhui Li ; Cibo Huang…
• 关键词：rheumatoid arthritis ; proteomic analysis ; orbitrap ; ficolin ; 2
• 刊名：Inflammation
• 出版年：2014
• 出版时间：October 2014
• 年：2014
• 卷：37
• 期：5
• 页码：1459-1467
• 全文大小：555 KB
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• 作者单位：Yongjing Cheng (2)
  Yuling Chen (3)
  Xiaolin Sun (1)
  Yuhui Li (1)
  Cibo Huang (2)
  Haiteng Deng (3)
  Zhanguo Li (1)
  
  2. Department of Rheumatology and Immunology, Beijing Hospital, 1 Dahua Road, 100730, Beijing, China
  3. School of Life Sciences, Tsinghua University, 100084, Beijing, China
  1. Department of Rheumatology and Immunology, People’s Hospital, Peking University, 11 Xizhimen South St., 100044, Beijing, China
  
• ISSN：1573-2576

文摘

The aim of this study was to find serum biomarkers of rheumatoid arthritis (RA) by high-resolution proteomic analysis. Low-abundance proteins from pooling serum sample of early RA patients and healthy controls were enriched using ProteoMiner?enrichment kits. The enriched proteins were separated on SDS-PAGE, digested by trypsin, labeled with tandem mass tag (TMT) reagents, and desalted by C18 stage tip column. Then, the labeled peptides were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with nano-LC combined with Orbitrap Q Exactive mass spectrometer, and experiments were carried out three times using different specimens, and differentially expressed proteins were screened by intensity ratios of identified peptides. Enzyme-linked immunosorbent assays (ELISAs) were performed to confirm differentially expressed proteins. Twenty-six proteins were found differentially expressed in RA serum by high-resolution proteomic analysis. Among them, levels of thrombospondin-1, ficolin-2, isoform 10 of fibronectin, and apolipoprotein E were higher in RA patients than in healthy controls (RA/healthy control (HC)?≥-.5, p--.05), while levels of angiotensinogen, paraoxonase/arylesterase 1, isoform E of proteoglycan 4, and plasminogen were significantly lower (RA/HC?≤-.67, p--.05). Further study by ELISA showed a higher level of ficolin-2 in the serum of RA patients compared to healthy controls; the level of ficolin-2 was found to be positively correlated with swollen joint counts (SJCs), disease activity score (DAS28), rheumatoid factor, and IgM in RA patients and with DAS28 and IgM in early RA patients through statistical analysis. The results of this study suggest that ficolin-2, as a newly screened biomarker by high-resolution quantitative proteomic analysis, offers the potentiality to become a diagnostic or disease evaluation tool in RA.