Pap T, Shigeyama Y, Kuchen S, Fernihough J K, Simmen B, Gay R E, Billingham M, Gay S
WHO Collaborating Center for Molecular Biology and Novel Therapeutic Strategies for Rheumatic Diseases, University Hospital, Zurich, Switzerland.
Arthritis Rheum. 2000 Jun;43(6):1226-32. doi: 10.1002/1529-0131(200006)43:6<1226::AID-ANR5>3.0.CO;2-4.
To study the expression of messenger RNA (mRNA) for different membrane-type matrix metalloproteinases (MT-MMPs) and compare their expression pattern in rheumatoid arthritis (RA) and normal synovium.
Polymerase chain reaction (PCR) with specific primers was performed to analyze the presence of MT1-, MT2-, MT3-, and MT4-MMP in synovial tissue and synovial fibroblasts from 10 patients with RA and 4 subjects without arthritis. In addition, in situ hybridization with digoxigenin-labeled RNA probes was used to investigate the expression pattern of MT-MMPs in the synovium of these subjects. MT-MMP-expressing cells were characterized by immunohistochemical double labeling with anti-CD68 monoclonal antibodies.
Reverse transcription-PCR revealed the expression of MT1-, MT2-, MT3-, and MT4-MMP mRNA in all tissues and cell cultures examined. However, in situ hybridization showed considerable differences in the expression pattern of the different MT-MMPs in RA synovium. MT1- and MT3-MMP mRNA were highly expressed in both the lining and the sublining layer, with more intense staining in the lining. Immunohistochemical double labeling demonstrated the presence of mRNA for MT1-MMP in fibroblasts and macrophages, as well as in osteoclast-like cells at sites of bone resorption. Expression of MT3-MMP mRNA was seen in fibroblasts and some macrophages. Expression of MT2- and MT4-MMP was characterized by staining of only a few CD68-negative fibroblasts, and no differences could be found between the lining and sublining. Normal synovial samples showed only limited staining for all MT-MMPs.
Our results indicate a role for MT1-MMP not only in the matrix degradation by fibroblasts, but also in osteoclast-mediated bone resorption in RA. Given the ability of MT1-MMP to activate MMP-2 and MMP-13, the findings also point to a cooperation between fibroblasts and macrophages in degrading cartilage and bone. While MT3-MMP is also intensely expressed in RA synovium, MT2- and MT4-MMP appear not to be involved in rheumatoid joint destruction.
研究不同膜型基质金属蛋白酶（MT - MMPs）信使核糖核酸（mRNA）的表达，并比较其在类风湿关节炎（RA）和正常滑膜中的表达模式。
采用特异性引物进行聚合酶链反应（PCR），分析10例RA患者和4例无关节炎受试者的滑膜组织及滑膜成纤维细胞中MT1 -、MT2 -、MT3 -和MT4 - MMP的存在情况。此外，使用地高辛标记的RNA探针进行原位杂交，以研究这些受试者滑膜中MT - MMPs的表达模式。通过抗CD68单克隆抗体免疫组化双重标记对表达MT - MMP的细胞进行鉴定。
逆转录 - PCR显示在所检测的所有组织和细胞培养物中均有MT1 -、MT2 -、MT3 -和MT4 - MMP mRNA的表达。然而，原位杂交显示RA滑膜中不同MT - MMPs的表达模式存在显著差异。MT1 -和MT3 - MMP mRNA在衬里层和衬里下层均高表达，衬里层染色更强。免疫组化双重标记显示，在成纤维细胞、巨噬细胞以及骨吸收部位的破骨细胞样细胞中均存在MT1 - MMP的mRNA。MT3 - MMP mRNA在成纤维细胞和一些巨噬细胞中表达。MT2 -和MT4 - MMP的表达表现为仅少数CD68阴性成纤维细胞染色，且衬里层和衬里下层之间未发现差异。正常滑膜样本中所有MT - MMPs的染色均有限。
我们的结果表明MT1 - MMP不仅在成纤维细胞介导的基质降解中起作用，而且在RA的破骨细胞介导的骨吸收中也起作用。鉴于MT1 - MMP具有激活MMP - 2和MMP - 13的能力，这些发现还表明成纤维细胞与巨噬细胞在降解软骨和骨方面存在协同作用。虽然MT3 - MMP在RA滑膜中也强烈表达，但MT2 -和MT4 - MMP似乎不参与类风湿关节破坏。
