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）的降解是破坏类风湿肌关节软骨（RA）的重要步骤，而基因金属蛋白酶（MMP）基因家族的成员参与了降解。在演讲研究中，我们表征了某些MMP物种的生化特性，并检查了它们在RA关节中软骨破坏的功能。此外，金属蛋白酶3（TIMP-3）可诱导药物的合成MMP抑制剂和组织抑制剂是通过与制药公司的合作开发的。主要结果如下：（1）。在TIMP-2存在的情况下加速了由膜型MMP（MT1-MMP）介导的ProMP-2的激活，因此TIMP-2被认为对于在细胞膜上有效激活Prommp-2是必不可少的。我们还证明了MT1-MMP在盘间域中的三个位点切割Aggrecan，MT3-MMP降解了ECM组件，例如III型胶原蛋白和脂肪蛋白。 o…另一方面，TIMP-1并未抑制MT1-MMP和MT3-MMP的活动。 MT-MMP被证明是从用姜黄蛋白A处理的肿瘤细胞的细胞膜中脱落的。（2）。在RA滑膜组织中有效激活了Prommp-2，其激活比与MT1，2、3-MMP之间的MT1-MMP的表达水平直接相关。 MT1-MMP在RA滑膜的衬里细胞中表达，并通过衬里细胞层中的原位zym摄影检测到明胶活性。（3）。通过三明治酶免疫测定分配了滑液中MMP-1、2、3、7、7、8、9、13和TIMP-1、2的稳态水平时，RA的MMP 1、2、3、8、9和TIMP-1的水平显着高于OA。 RA的摩尔比显着高于OA，并且与MMP/TIMP摩尔比的直接相关性检测到金属蛋白酶活性，这表明有利于蛋白酶的失衡。（4）。通过与一家制药公司的合作，我们开发了对MT1-MMP更具选择性的合成抑制剂，我们还证明，五角星多硫酸盐会刺激RA滑膜成纤维细胞，以通过转录后水平产生TIMP-3。较少的