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类风湿关节炎滑膜成纤维样细胞的蛋白质组学分析。

Proteomic analysis of synovial fibroblast-like synoviocytes from rheumatoid arthritis.

机构信息

School of Medicine, Nanjing University, Nanjing, China.

出版信息

Clin Exp Rheumatol. 2013 Jul-Aug;31(4):552-8. Epub 2013 Jun 5.

Abstract

OBJECTIVES

The purpose of this study was to identity protein expression patterns of fibroblast-like synoviocytes (FLSs) derived from the synovial tissue of patients with rheumatoid arthritis (RA) and osteoarthritis.

METHODS

Two-dimensional gel electrophoresis (2-DE) in combination with matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF-MS) was used to visualise and identify differential cellular protein expression profiles in FLSs between RA and control groups. Western-blot analysis was performed to further verify selected differentially-expressed proteins.

RESULTS

A total of 1633 and 1603 protein spots were examined in synovial FLSs of RA patients and controls, respectively. Ninety-two spots in the RA group were statistically over- or under-expressed compared with controls. Among them, 33 proteins over-expressed by more than 3-fold were then identified by MALDI-TOF-MS analysis. These proteins included enzymatic and structural proteins (e.g. PKM1/M2, α-enolase, ERp60, lamin-A/C), signal transduction proteins (e.g. annexin 11, peroxiredoxin 1, TrpRS), heat-shock/chaperone proteins (e.g. TCP-1, GRP75, HspB5, Bip) and some unknown protein species. Three proteins, namely α-enolase, GRP75 and PKM2, were verified by Western blot and the results were found to be consistent with proteomic analysis.

CONCLUSIONS

The differentially expressed proteins identified in RA synovial FLSs might be candidate RA-associated proteins and may prove to be promising diagnostic indicators or new therapeutic targets for RA.

摘要

目的

本研究旨在鉴定类风湿关节炎(RA)和骨关节炎患者滑膜组织来源的成纤维样滑膜细胞(FLS)的蛋白表达模式。

方法

采用二维凝胶电泳(2-DE)结合基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术,可视化和鉴定 RA 组和对照组 FLS 中差异表达的细胞蛋白表达谱。采用 Western-blot 分析进一步验证选定的差异表达蛋白。

结果

在 RA 患者和对照组的滑膜 FLS 中分别检测到 1633 和 1603 个蛋白点。RA 组中有 92 个蛋白点的表达与对照组相比统计学上过度或不足。其中,MALDI-TOF-MS 分析鉴定出 33 个表达超过 3 倍的蛋白。这些蛋白包括酶和结构蛋白(如 PKM1/M2、α-烯醇酶、ERp60、层粘连蛋白-A/C)、信号转导蛋白(如 annexin 11、过氧化物还原酶 1、TrpRS)、热休克/伴侣蛋白(如 TCP-1、GRP75、HspB5、Bip)和一些未知的蛋白种类。通过 Western blot 验证了 3 种蛋白,即α-烯醇酶、GRP75 和 PKM2,结果与蛋白质组学分析一致。

结论

RA 滑膜 FLS 中差异表达的蛋白可能是 RA 相关的候选蛋白,可能成为有前途的 RA 诊断指标或新的治疗靶点。

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