New collagenolytic enzymes cascade identified at the pannus-hard tissue junction in rheumatoid arthritis: Destruction from above
New collagenolytic enzymes cascade identified at the pannus-hard tissue junction in rheumatoid arthritis: Destruction from above
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DOI：
10.1016/s0945-053x(98)90110-x
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发表时间：
1998-12-01
期刊：
影响因子：
6.9
通讯作者：
Seiki, M
中科院分区：
文献类型：
--
作者：
Konttinen, YT;Ceponis, A;Seiki, M
我们的目的是研究基质金属蛋白酶2 （matrix metalloproteinase-2, MMP-2）的胶原溶解潜能和定位与其调节蛋白膜型MT1-MMP和金属蛋白酶组织抑制剂-2的关系；TIMP-2在类风湿关节炎（RA）中的表达。为此，我们纯化了MMP-2、MMP-8、MMP-9和间质I、II和III型胶原；SDS-PAGE/密度法测定胶原酶活性；zymography;西方墨点法;逆转录酶聚合酶链反应；原位杂交；免疫荧光、ABC、ABC- apap双免疫染色。MMP-2降解人II型胶原几乎与MMP-8一样有效，而MMP-9不裂解人II型胶原。在滑膜组织中，在成纤维细胞和巨噬细胞样细胞、基质细胞和血管内皮中发现了MT1-MMP、TIMP-2和MMP-2。MT1-MMP、TIMP-2和MMP-2在浸润性输卵管交界面泛细胞中表达强烈，但软硬组织（软骨和/或骨）交界面“上方”和“下方”染色较弱和/或阳性细胞较少。类风湿性滑膜组织提取物含有62/59 kDa的蛋白水解活性MMP-2和43 kDa的MT1-MMP，但没有游离的TIMP-2。这些结果表明，三元MT1-MMP/TIMP-2/MMP-2复合物的组分在正常滑膜衬里及其在透明关节软骨上的病理延伸中共表达。MMP-2可能参与正常内膜的重塑，也可能定位/聚焦于关节炎组织破坏关键部位的泛细胞。
Our aim was to investigate the collagenolytic potential and localization of matrix metalloproteinase-2 (MMP-2) in relation to its regulatory proteins membrane type MT1-MMP and tissue inhibitor of metalloproteinases-2; (TIMP-2) in rheumatoid arthritis (RA). For this purpose, we have used purification of MMP-2, MMP-8, MMP-9 and interstitial type I, II and III collagens; SDS-PAGE/densitometric collagenase activity assay; zymography; Western blotting; reverse transcriptase polymerase chain reaction; in situ hybridization; and immunofluorescence, ABC, ABC-APAAP double immunostainings. MMP-2 degraded human type II collagen almost as effectively as MMP-8, whereas MMP-9 did not cleave type II collagen. In synovial tissue, MT1-MMP, TIMP-2 and MMP-2 were found in synovial lining in fibroblast- and macrophage-like cells, in stromal cells and in vascular endothelium. MT1-MMP, TIMP-2 and MMP-2 were strongly expressed in the pannocytes of the invasive pannus at the interface, but staining was weak and/or there were few positive cells both "above" and "below" the soft-to-hard tissue (cartilage and/or bone) interface. Rheumatoid synovial tissue extract contained proteolytically active 62/59 kDa MMP-2 and 43 kDa MT1-MMP, but no free TIMP-2. These results indicate that components of the ternary MT1-MMP/TIMP-2/MMP-2 complex are coexpressed in the normal synovial lining and in its pathological extension on the hyaline articular cartilage. MMP-2 may participate in the remodeling of the normal lining and also seems to be localized/focalized to pannocytes at a site critical for tissue destruction in arthritis.