Detection of high levels of 2 specific isoforms of 14-3-3 proteins in synovial fluid from patients with joint inflammation.
Detection of high levels of 2 specific isoforms of 14-3-3 proteins in synovial fluid from patients with joint inflammation.
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发表时间：
2007-08
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通讯作者：
R. Kilani;W. Maksymowych;A. Aitken;G. Boire;Y. St-Pierre;Yunyuan Li;A. Ghahary
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作者：
R. Kilani;W. Maksymowych;A. Aitken;G. Boire;Y. St-Pierre;Yunyuan Li;A. Ghahary
目的探讨关节炎症患者滑液中是否检测到14-3-3蛋白，以及14-3-3蛋白的亚型是什么(S)，并探讨其与关节炎性滑液中基质金属蛋白酶-1和基质金属蛋白酶-3的表达水平是否存在相关性。方法对2组滑膜和血清标本进行分析。对17例关节炎症患者的第一组SF样本进行了14-3-3ETA亚型的Western印迹分析。第二组12个匹配的血清和SF样本用相同的程序分析了14-3-3ETA、伽马、基质金属蛋白酶-1和基质金属蛋白酶-3。观察不同浓度的14-3-3-ETA对成纤维细胞基质金属蛋白酶-1的刺激作用。结果在检测的7种14-3-3亚型(β、γ、epsilon、eta、sigma、theta和zeta)中，只有两种亚型在炎症性关节疾病患者的SF标本中容易检测到。在炎症性SF和血清样本中，这些蛋白的水平明显高于对照组。这些蛋白的值与血清中检测到的类风湿性关节炎的两个生物标志物--基质金属蛋白酶-1和基质金属蛋白酶-3的水平密切相关。此外，在来自炎症性关节疾病患者的12份血清样本中，14-3-3 ETA水平显著高于健康人。结论在SF的7种不同亚型中只检测到2种(14-3-3ETA和伽马)，提示它们是炎症部位的特异性异构体，这与正常血清中几乎检测不到的这些亚型有所区别。ETA异构体对基质金属蛋白酶-1的刺激作用解释了它与在这些样本中看到的基质金属蛋白酶-1水平的相关性。
OBJECTIVE To investigate whether 14-3-3 proteins were detectable in synovial fluid (SF) of patients with inflamed joints, and if so, what isoform(s); and to examine whether there was a correlation between the levels of these proteins and those of MMP-1 and MMP-3 in the same samples. METHODS In general, 2 sets of synovial and serum samples were analyzed. The first set of 17 SF -samples from patients with inflamed joints were analyzed for 14-3-3 eta isoform by Western blot. The second set of 12 matching serum and SF samples were analyzed for 14-3-3 eta, gamma, MMP-1, and MMP-3 by the same procedure. The MMP-1 stimulatory effect of various concentrations of 14-3-3 eta in cultured fibroblasts was then evaluated. RESULTS We found that of the seven 14-3-3 isoforms tested (beta, gamma, epsilon, eta, sigma, Theta, and zeta), the levels of only 2 isoforms, eta and gamma, were easily detectable in SF samples from patients with inflammatory joint diseases. The levels of these proteins were significantly higher in inflammatory SF and serum samples relative to controls. The values of these proteins correlated strongly with the levels of MMP-1 and MMP-3, 2 biomarkers for rheumatoid arthritis, detected in sera. Further, the level of 14-3-3 eta was significantly higher in a pool of 12 serum samples from patients with inflammatory joint disease than those from healthy individuals. CONCLUSION Detection of only 2 (14-3-3 eta and gamma) out of 7 different isoforms in SF suggests they are specific to the site of inflammation, and that distinguishes them from barely detectable levels of these isoforms found in normal serum. The MMP-1 stimulatory effect of the eta isoform explains its correlation with MMP-1 levels seen in these samples.