Studies on the destruction of articular cartilage by matrix metalloproteinases

基质金属蛋白酶破坏关节软骨的研究

• 批准号: 07457049
• 负责人: OKADA Yasunori
• 金额: $ 4.93万
• 依托单位: Kanazawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457049/
• 关键词: Matrix metalloproteinase; Rheumatoid arthritis; Osteoarthritis; Cartilage destruction; Degradation; Tissue inhibitor; Cytokine; Activation; 細胞外マトリックス分解; 関節破壊

Matrix metalloproteinases (MMPs) are composed of at least 15 different molecules and their activities are strictly regulated by their common tissue inhibitors of metalloproteinases (TIMP-1,2,3,4). In the present studies, we have demonstrated that membrane-type 1 matrix metalloproteinase (MT1-MMP) is highly expressed in the human osteoarthritic cartilage showing a positive correlation with activation of proMMP-2 (progelatinase A). We have also revealed that MT1-MMP is an extracellular matrix (ECM) -degrading proteinase capable of digesting interstitial collagens and aggrecan as well as an activator of proMMP-2. In osteoarthritis and rheumatoid arthritis, cartilage matrix protein and SPARC were overexpressed, suggesting that it may be a tissue reaction secondary to the degradation of the ECM components by MMPs expressed in the cartilages. Actually, TGF-beta which stimulates ECM production in various mesenchymal cells was released after degradation of the TGF-beta/decorin complex by MMPs. To determine the production levels, sandwich enzyme immunoassays for MMP-7 and MMP-8 were also developed. Research projects on the mechanisms of MT1-MMP inhibition by TIMP-2 and expression of TIMP-1,2,3,4 in osteoarthritic and rheumatoid arthritic cartilages are now under way.

基质金属蛋白酶（Matrix metalloproteinases，MMPs）由至少15种不同的分子组成，其活性受其共同的组织金属蛋白酶抑制剂（tissue inhibitor of metalloproteinases，TIMP-1，2，3，4）的严格调控。在本研究中，我们已经证明，膜型1基质金属蛋白酶（MT 1-MMP）是高表达的人骨关节炎软骨显示与MMP-2（明胶酶A）的激活呈正相关。我们还发现，MT 1-MMP是一种细胞外基质（ECM）降解蛋白酶，能够消化间质胶原和聚集蛋白聚糖以及proMMP-2的激活剂。在骨关节炎和类风湿性关节炎中，软骨基质蛋白和ECM过度表达，表明这可能是继发于软骨中表达的MMP降解ECM组分的组织反应。实际上，在各种间充质细胞中刺激ECM产生的TGF-β在TGF-β/核心蛋白聚糖复合物被MMP降解后释放。为了确定生产水平，还开发了MMP-7和MMP-8的夹心酶免疫测定法。TIMP-2对MT 1-MMP的抑制机制以及TIMP-1、2、3、4在骨关节炎和类风湿关节炎软骨中的表达的研究正在进行中。

项目成果

Ohashi K., Kawai R., Hara M., Okada Y., Tachibana S.and Ogura Y.: "Increased matrix metalloproteinases as possible cause of osseo-articular tissue destruction in long term hemodailysis and beta2-microglobulin amyloidosis." Virchows Archiv. 428. 37-46 (199

Ohashi K.、Kawai R.、Hara M.、Okada Y.、Tachibana S.和 Ogura Y.：“基质金属蛋白酶的增加可能是长期血液透析和 β2-微球蛋白淀粉样变性中骨关节组织破坏的原因。”

Okimura A., Okada Y., Makihira S., Pan H., Yu L., Tanne K., Imai K., Yamada H., Kawamoto T., Noshiro M., Yan W.and Kato Y.: "Cartilage-matrix protein (CMP) synthesis is enhanced in arthritic cartilage." Arthritis Rheum. (in press). (1997)

Okimura A.、Okada Y.、Makihira S.、Pan H.、Yu L.、Tanne K.、Imai K.、Yamada H.、Kawamoto T.、Noshiro M.、Yan W.和 Kato Y.：“软骨

Imai K.: "Degradation of decorin by matrix metalloproteinases. Identification of the cleavage sites, kinetic analyses and transforming growth factor-βl release." Biochem. J.(in press). (1997)

Imai K.：“基质金属蛋白酶对核心蛋白聚糖的降解。切割位点的鉴定、动力学分析和转化生长因子-β1 释放。Biochem J.（出版中）。”

Matsuki H., Fujimoto N., Iwata K., Knauper V., Okada Y.and Hayakawa T.: "A one-step sandwich enzyme immunoassay for human matrix metalloproteinase 8 (neutrophil collagenase) using monoclonal antibodies" Clin.Chim.Acta. 244. 129-143 (1996)

Matsuki H.、Fujimoto N.、Iwata K.、Knauper V.、Okada Y. 和 Hayakawa T.：“使用单克隆抗体对人基质金属蛋白酶 8（中性粒细胞胶原酶）进行一步夹心酶免疫测定” Clin.Chim.Acta

Ito M., Masuda K., Ito Y., Akizawa T., Yoshioka M., K.Imai, Okada Y., Sato H.and Seiki M.: "Purification and refolding of recombinant human proMMP-7 (pro-matrilysin) expressed in Echelichia coil and its characterization." J.Biochem. 119. 667-673 (1996)

Ito M.、Masuda K.、Ito Y.、Akizawa T.、Yoshioka M.、K.Imai、Okada Y.、Sato H. 和 Seiki M.：“重组人 proMMP-7（pro-matrilysin）的纯化和重折叠