A novel approach to measure the contribution of matrix metalloproteinase in the overall net proteolytic activity present in synovial fluids of patients with arthritis
A novel approach to measure the contribution of matrix metalloproteinase in the overall net proteolytic activity present in synovial fluids of patients with arthritis
复制标题
一种测量基质金属蛋白酶在关节炎患者滑液中总体净蛋白水解活性中的贡献的新方法
作者：
N. Simard;G. Boire;A. D. de Brum;Y. St
尽管进行了数十年的研究，但只有非常有限的基质金属蛋白酶 (MMP) 抑制剂在关节炎的临床试验中取得了成功。与这种失败相关的核心问题之一可能是我们无法监测关节中蛋白酶的局部活性，因为细胞外基质的完整性是由蛋白酶抑制剂的非共价、1:1化学计量结合与酶的活化形式的催化位点之间的平衡产生的。在目前的工作中，我们通过流式细胞术测量了从 95 名骨关节炎和各种形式的炎性关节炎（包括类风湿性关节炎、脊柱关节病和慢性幼年关节炎）患者收集的滑液 (SF) 中的净蛋白水解活性。我们发现炎症性关节炎患者的 SF 蛋白水解活性水平显着高于骨关节炎患者。此外，炎症性关节炎患者的总体活动与浸润白细胞的数量和C反应蛋白的血清水平呈正相关。在骨关节炎患者中没有发现这种相关性。 MMP 家族的成员对 SF 中的蛋白水解活性有显着贡献。小分子量 MMP 抑制剂确实能有效抑制 SF 的蛋白水解活性，但其效果在患者之间差异很大。有趣的是，在蛋白水解活性非常高的患者中，MMP 的贡献下降，这是由于 MMP-1 组织抑制剂摩尔过量和其他蛋白水解酶的贡献增加所致。这些结果强调了与关节炎有关的 MMP 的多样性，并且从临床角度来看，提出了一种有趣的替代方案来测试新型蛋白酶抑制剂治疗关节炎的潜力。
Despite decades of research, only a very limited number of matrix metalloproteinase (MMP) inhibitors have been successful in clinical trials of arthritis. One of the central problems associated with this failure may be our inability to monitor the local activity of proteases in the joints since the integrity of the extracellular matrix results from an equilibrium between noncovalent, 1:1 stoichiometric binding of protease inhibitors to the catalytic site of the activated forms of the enzymes. In the present work, we have measured by flow cytometry the net proteolytic activity in synovial fluids (SF) collected from 95 patients with osteoarthritis and various forms of inflammatory arthritis, including rheumatoid arthritis, spondyloarthropathies, and chronic juvenile arthritis. We found that SF of patients with inflammatory arthritis had significantly higher levels of proteolytic activity than those of osteoarthritis patients. Moreover, the overall activity in inflammatory arthritis patients correlated positively with the number of infiltrated leukocytes and the serum level of C-reactive protein. No such correlations were found in osteoarthritis patients. Members of the MMP family contributed significantly to the proteolytic activity found in SF. Small-molecular-weight MMP inhibitors were indeed effective for inhibiting proteolytic activity in SF, but their effectiveness varied greatly among patients. Interestingly, the contribution of MMPs decreased in patients with very high proteolytic activity, and this was due both to a molar excess of tissue inhibitor of MMP-1 and to an increased contribution of other proteolytic enzymes. These results emphasize the diversity of the MMPs involved in arthritis and, from a clinical perspective, suggest an interesting alternative for testing the potential of new protease inhibitors for the treatment of arthritis.
影响因子：
4.1
作者：
Vincenti, MP;Coon, CI;Brinckerhoff, CE
通讯作者：
Brinckerhoff, CE
影响因子：
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作者：
Ahrens, D;Koch, AE;Niedbala, MJ
通讯作者：
Niedbala, MJ