Expression of matrix metalloproteinase 9 (96-kd gelatinase B) in human rheumatoid arthritis
Expression of matrix metalloproteinase 9 (96-kd gelatinase B) in human rheumatoid arthritis
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DOI：
10.1002/art.1780390919
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发表时间：
1996-09-01
影响因子：
--
通讯作者：
Niedbala, MJ
中科院分区：
文献类型：
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作者：
Ahrens, D;Koch, AE;Niedbala, MJ
目标。采用特异性单克隆抗体检测类风湿性关节炎（RA）、炎性关节炎（IA）和骨关节炎（OA）患者滑膜液（SF）、血浆和滑膜组织中基质金属蛋白酶9/gelantinase B （MMP-9）的表达。用明胶酶谱法评估RA、IA和OA患者SF和血浆中的明胶溶解活性。小鼠单克隆抗血清277.13可选择性识别人MMP-9的可溶性潜伏形式，使用捕获夹心酶联免疫吸附试验（ELISA）定量测定患者滑膜积液、血浆和滑膜组织中的MMP-9水平。分析了51份SF样本（RA 31份，OA 9份，IA 11份）。用MMP-9特异性抗血清研究了MMP-9在RA、OA和正常滑膜中的免疫定位。与OA样本相比，RA样本滑膜积液中MMP-9抗原水平升高67倍。此外，尽管IA滑膜积液中的MMP-9抗原水平比RA样品低2.7倍，但比OA样品高34倍。这些数据表明MMP-9水平升高与炎症性关节炎有关。在RA和IA样品中明显存在明显的92-kd明胶溶解活性（被EDTA特异性抑制），但在OA样品中未观察到活性。免疫捕获ELISA法检测86份血浆样本（RA 17份，IA 9份，正常人60份）MMP-9抗原水平，RA血浆中MMP-9抗原水平比正常人高7倍。与OA滑膜组织相比，RA滑膜组织提取物显示MMP-9抗原水平升高。免疫定位研究表明，MMP-9在RA滑膜浸润性白细胞（中性粒细胞和巨噬细胞）、内皮细胞和滑膜成纤维细胞中表达。与OA SF相比，RA和IA SF中潜在的MMP-9和/或MMP-9金属蛋白酶组织抑制剂1 （TIMP-1）复合物升高。此外，与正常对照血浆相比，RA血浆中MMP-9升高。滑膜组织MMP-9抗原水平在RA与OA中也升高。MMP-9在RA滑膜内的组织分布局限于炎症部位，包括滑膜表面衬里细胞、内皮细胞和白细胞。综上所述，这些观察结果表明，结缔组织的更新是MMP活性超过TIMP作用的结果，而TIMP作用在侵入性肠膜、关节周围组织或SF中。进一步的研究，如在本研究中使用的，将有助于阐明许多不同的酶和抑制剂在破坏性关节病中的作用。
Objective. To determine the expression of matrix metalloproteinase 9/gelantinase B (MMP-9) in synovial fluid (SF), plasma, and synovial tissue from individuals with rheumatoid arthritis (RA), inflammatory arthritis (IA), and osteoarthritis (OA), using specific monoclonal antibody reagents.Methods. Gelatinolytic activity in the SF and plasma of patients with RA, IA, and OA was assessed by gelatin zymography. A mouse monoclonal antiserum, 277.13, which selectively recognizes soluble latent forms of human MMP-9, was used to quantitate MMP-9 levels in patient synovial effusions, plasma, and synovial tissue with a capture sandwich enzyme-linked immunosorbent assay (ELISA). Fifty-one SF samples (31 RA, 9 OA, 11 IA) were analyzed. Immunolocalization of MMP-9 in RA, OA and normal synovium was investigated using MMP-9-specific antisera.Results. MMP-9 antigen levels in synovial effusions were elevated 67-fold in RA samples compared with OA samples. In addition, although MMP-9 antigen levels in IA synovial effusions were 2.7-fold less than the values in RA samples, they were elevated 34-fold over the values in OA samples. These data indicate an association between increased MMP-9 levels inflammatory arthritis. A predominant 92-kd gelatinolytic activity (specifically inhibited by EDTA) was evident in RA and IA samples, but no activity was observed in OA samples. Among 86 plasma samples (17 RA, 9 IA, 60 normal controls) analyzed for MMP-9 antigen levels by immunocapture ELISA, MMP-9 antigen levels were elevated 7-fold in RA plasma compared with normal plasma. RA synovial tissue extracts demonstrated elevated levels of MMP-9 antigen compared with OA synovial tissue. MMP-9 immunolocalization studies demonstrated expression in infiltrating leukocytes (neutrophils and macrophages), endothelial cells, and synovial fibroblasts in RA synovium.Conclusion. Latent MMP-9 and/or MMP-9-tissue inhibitor of metalloproteinases 1 (TIMP-1) complexes are elevated in RA and IA SF compared with OA SF. In addition, MMP-9 is increased in RA plasma versus normal control plasma. Synovial tissue levels of MMP-9 antigen are also elevated in RA versus OA. The tissue distribution of MMP-9 within RA synovium is localized to sites of inflammation comprising surface synovial lining cells, endothelium, and leukocytes. Taken together, these observations suggest that connective tissue turnover occurs as a result of excessive MMP activity over TIMP action in the invading pannus, periarticular tissue, or SF. Further studies such as those used in the present investigation will help elucidate the role of a number of different enzymes and inhibitors in the destructive arthropathies.