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Molecular Pathology of Cartilage destruction in Rheumatoid Arthritis

类风湿关节炎软骨破坏的分子病理学

基本信息

批准号：
10470051
负责人：
OKADA Yasunori
金额：
$ 8.13万
依托单位：
Keio University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

项目摘要

Degradation of the extracellular matrix (ECM) is an essential step to the destruction of articular cartilage in rheumatoid athritis (RA), and members of the matrix metalloproteinase (MMP) gene family are involved in the degradation. In the present studies, we characterized the biochemical properties of some MMP species and examined their functions in the destruction of the cartilage in RA joints. In addition, synthetic MMP inhibitors and tissue inhibitor of metalloproteinases 3 (TIMP-3)-inducible drug were developed by collaboration with pharmaceutical companies. The major results are as follows:(1). Activation of proMMP-2 mediated by membrane-type1 MMP (MT1-MMP) was accelerated in the presence of TIMP-2, and thus TIMP-2 was considered to be essential to the efficient activation of proMMP-2 on the cell membranes. We also demonstrated that MT1-MMP cleaves aggrecan at three sites in the interglobular domain and MT3-MMP degrades the ECM components such as type III collagen and aggrecan. O … More n the other hand, the activities of MT1-MMP and MT3-MMP were not inhibited by TIMP-1. MT-MMP was shown to be shed from the cell membranes of tumor cells treated with concanvalin A.(2). ProMMP-2 was efficiently activated in RA synovial tissues and its activation ratio directly correlated with the expression levels of MT1-MMP among MT1, 2, 3-MMPs. MT1-MMP was expressed in the lining cells of RA synovial membrane and gelatinolytic activity was detected by in situ zymography in the lining cell layer.(3). When steady state levels of MMP-1, 2, 3, 7, 8, 9, 13 and TIMP-1, 2 in synovial fluids were assayed by sandwich enzyme immunoassays, the levels of MMP 1, 2, 3, 8, 9 and TIMP-1 were significantly higher in RA than in OA. The molar ratio was significantly higher in RA than in OA, and metalloproteinase activity was detected with a direct correlation to MMP/TIMP molar ratio, suggesting an imbalance in favor of proteinase.(4). By collaboration with a pharmaceutical company, we developed synthetic inhibitors more selective to MT1-MMP, We also demonstrated that pentosan polysulfate stimulates RA synovial fibroblasts to produce TIMP-3 by the posttranscriptional level. Less

细胞外基质（ECM）的降解是类风湿性关节炎（RA）关节软骨破坏的重要步骤，基质金属蛋白酶（MMP）基因家族的成员参与了降解。在本研究中，我们表征了一些 MMP 物种的生化特性，并检查了它们在破坏 RA 关节软骨中的功能。此外，还与制药公司合作开发了合成MMP抑制剂和金属蛋白酶3（TIMP-3）诱导药物的组织抑制剂。主要研究结果如下： (1)． TIMP-2 存在时，膜型 1 MMP (MT1-MMP) 介导的 proMMP-2 的激活会加速，因此 TIMP-2 被认为对于细胞膜上 proMMP-2 的有效激活至关重要。我们还证明 MT1-MMP 在球间结构域的三个位点裂解聚集蛋白聚糖，MT3-MMP 降解 ECM 成分，如 III 型胶原和聚集蛋白聚糖。另一方面，MT1-MMP 和 MT3-MMP 的活性不受 TIMP-1 的抑制。经伴刀豆球蛋白 A 处理的肿瘤细胞的细胞膜显示 MT-MMP 脱落。(2)。 ProMMP-2在RA滑膜组织中被有效激活，其激活率与MT1、2、3-MMP中MT1-MMP的表达水平直接相关。 MT1-MMP在RA滑膜衬里细胞中表达，并通过原位酶谱法检测衬里细胞层的明胶分解活性。(3)．当通过夹心酶免疫分析法测定滑液中MMP-1、2、3、7、8、9、13和TIMP-1、2的稳态水平时，RA中MMP-1、2、3、8、9和TIMP-1的水平显着高于OA。 RA 中的摩尔比显着高于 OA，并且检测到的金属蛋白酶活性与 MMP/TIMP 摩尔比直接相关，表明存在有利于蛋白酶的不平衡。(4)。通过与制药公司合作，我们开发了对 MT1-MMP 更具选择性的合成抑制剂，我们还证明多硫酸戊聚糖可刺激 RA 滑膜成纤维细胞在转录后水平产生 TIMP-3。较少的