类风湿关节炎滑膜成纤维样细胞的蛋白质组学研究

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

目的研究类风湿关节炎(RA)和骨性关节炎(OA)关节滑膜成纤维样细胞(FLS)蛋白质组的表达差异,分析差异蛋白在RA关节滑膜增生中的作用。方法原代培养关节滑膜FLS,抽提细胞总蛋白并定量,通过高分辨双向电泳(2-DE)对RA和OA的滑膜FLS进行蛋白分离,找出其中表达差异的蛋白质点并进行质谱(MS)分析鉴定。结果 OA及RA滑膜FLS的2-DE图谱上分别显示有1324和1147个蛋白质点。其中47个蛋白质点在OA或RA滑膜FLS中有3倍以上的量变;12个蛋白质点只存在于OA滑膜FLS;15个蛋白质点只存在于RA滑膜FLS。选择54个蛋白质点进行MS,共成功鉴定34个蛋白质点。其中包括在OA中表达上升的人异天冬氨酸甲基转移酶(PIMT)和PIR(PIR),仅在RA中表达的硫氧还蛋白1(TRX-1)这三个与关节滑膜增生相关的蛋白质。结论 PIMT和PIR在RA中表达降低及TRX-1在RA中表达升高可能是RA关节滑膜增生,引起关节软骨破坏的原因之一。

Objective Studies have found that the fibroblast-like synoviocytes(FLS)play an important role in the hyperplasia of synovium.However,which molecules of FLS caused the hyperplasia remains unclear.This study was performed to separate the differentially expressed proteins of FLS from the patients with rheumatoid arthritis(RA)or osteoarthritis(OA)by two-dimensional electrophoresis(2-DE),and found proteins associated with hyperplasia of FLS by mass spectrometry(MS).Methods Total proteins were extracted and quantified from the primary cultured FLS from patients of RA or OA.Proteins were separated by high-resolution 2-DE,and the differentially expressed proteins were identified by MS.Results There were 1147 protein spots from RA and 1324 protein spots from OA showed on 2-DE graphs,respectively.We had selected 54 protein spots for MS analysis,and 34 protein spots were successfully identified.We had selected the human protein isoaspartate methyltransferase(PIMT)and pirin(iron-binding nuclear protein,PIR)high expressed in FLS of OA,thioredoxin 1(TRX-1)only expressed in FLS of RA.These 3 proteins may be associated with hyperplasia of synovium.Conclusions The decreased expression of PIMT,PIR and increased expression of TRX-1 may be one of the reasons for hyperplasia of FLS in RA.