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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（extracellular matrix，ECM）的降解是类风湿关节炎（rheumatoid athlete，RA）关节软骨破坏的重要环节，基质金属蛋白酶（matrix metalloproteinase，MMP）基因家族成员参与了ECM的降解。在目前的研究中，我们的特点是一些MMP物种的生化特性，并检查它们的功能，在RA关节软骨的破坏。此外，与制药公司合作开发了合成MMP抑制剂和金属蛋白酶组织抑制剂3（TIMP-3）诱导型药物。主要结果如下：（1）.在TIMP-2的存在下，由膜型1 MMP（MT 1-MMP）介导的proMMP-2的活化被加速，因此TIMP-2被认为是proMMP-2在细胞膜上的有效活化所必需的。我们还证明了MT 1-MMP在球间结构域的三个位点切割聚集蛋白聚糖，MT3-MMP降解ECM组分，如III型胶原和聚集蛋白聚糖。O ...更多信息 TIMP-1对MT 1-MMP和MT3-MMP活性无明显抑制作用。MT-MMP从伴刀豆素A处理的肿瘤细胞的细胞膜上脱落。（二）、ProMMP-2在RA滑膜组织中被有效激活，其激活率与MT 1-MMP在MT 1，2，3-MMPs中的表达水平直接相关。MT 1-MMP表达于RA滑膜衬里层细胞，原位酶谱法检测衬里层细胞明胶酶活性。（三）、用双抗体夹心酶免疫法测定关节液中MMP-1、2、3、7、8、9、13和TIMP-1、2的稳态水平，发现RA组MMP-1、2、3、8、9和TIMP-1的水平显著高于OA组。RA中MMP/TIMP的摩尔比显著高于OA，且MMP/TIMP的摩尔比与金属蛋白酶活性呈正相关，提示蛋白酶的失衡。（四）、通过与制药公司合作，我们开发了对MT 1-MMP更具选择性的合成抑制剂，我们还证明了戊聚糖多硫酸酯通过转录后水平刺激RA滑膜成纤维细胞产生TIMP-3。少