Proteins related to the functions of fibroblast-like synoviocytes identified by proteomic analysis

• Published in: Clinical and experimental rheumatology 2012-03, Vol.30 (2), p.213-221
• Main Authors: ZHANG, H, FAN, L. Y, ZONG, M, SUN, L. S, LU, L
• Format: Article
• Language: English
• Subjects: Adult; Aged; Arthritis, Rheumatoid - genetics; Arthritis, Rheumatoid - metabolism; Arthritis, Rheumatoid - pathology; Biological and medical sciences; Blotting, Western; Carrier Proteins - metabolism; Cells, Cultured; Diseases of the osteoarticular system; Electrophoresis, Gel, Two-Dimensional; Female; Fibroblasts - metabolism; Fibroblasts - pathology; Humans; Inflammatory joint diseases; Male; Medical sciences; Middle Aged; Nuclear Proteins - metabolism; Osteoarthritis, Knee - genetics; Osteoarthritis, Knee - metabolism; Osteoarthritis, Knee - pathology; Protein D-Aspartate-L-Isoaspartate Methyltransferase - metabolism; Proteins - genetics; Proteins - metabolism; Proteomics - methods; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Synovial Membrane - metabolism; Synovial Membrane - pathology; Tandem Mass Spectrometry; Thioredoxins - metabolism
• ISSN: 0392-856X; 1593-098X

It is well known that the fibroblast-like synoviocytes (FLS) play a key role in pathogenesis of rheumatoid arthritis (RA). This study was performed to separate the differentially expressed proteins of FLS from the patients with RA or osteoarthritis (OA) by two-dimensional electrophoresis (2-DE), and found proteins associated with the functions of FLS by mass spectrometry (MS). Total proteins were extracted and quantified from the primary cultured FLS from patients of RA (n=8) or OA (n=6). Proteins were separated by high-resolution 2-DE, and identified the differentially expressed proteins by MS. Western blot analyses was used to validated the expression of candidate proteins. The mRNA of these proteins was detected by semi-quantitative fluorescent PCR. There are 1147 protein spots from RA and 1324 protein spots from OA showed on 2-DE graphs, respectively. We have selected 84 protein spots for MS analysis, and 27 protein spots were successfully identified. We have found that protein isoaspartyl methyltransferase (PIMT) and pirin (iron-binding nuclear protein, PIR) with lower expression in RA, and thioredoxin 1(Trx-1) only expressed in RA may be associated with functions of FLS. Western Blot confirmed the expression of PIMT and pirin lower in RA, and Trx-1 expressed only in RA. The results of semi-quantitative fluorescent PCR are also consistent with 2-DE graphs. PIMT, pirin and Trx-1 affect the functions of FLS in some style and can be the drug targets of RA.