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The pathophysiological significance of proteases in patients with chronic inflammatory joint disease

蛋白酶在慢性炎症性关节病患者中的病理生理意义

基本信息

批准号：
12671421
负责人：
NAKANO Shunji
金额：
$ 1.92万
依托单位：
The University of Tokushima
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671421/
关键词：
proteas rheumatoid arthritis osthoarthritis synovial fluid synovium トロンビントリプシンカテプシンB

项目摘要

To clarify the pathophysiological significance of proteases in RA and OA, we investigated what kinds of proteases were specifically activated in both synovial fluid and synovial tissue of RA ＆ OA patients , and effects of serine proteases on release of inflammatory cytokines from synovial cells.We obtained the following fmdings. (I) When the protease substrates were incubated with SF supernatants, thrombin was the most preferentially hydrolyzed while trypsin substrate and cathepsin B substrate were hydrolyzed to some degree. In the case of SF cell extracts, cathepsin B substrate was the most preferentially hydrolyzed while the other substrates were hardly hydrolyzed. In the case of synovial tissue extracts, cathepsin B substrate was the most preferentially hydrolyzed while trypsin substrate was hydrolyzed to some degree. Protease activities were in general significantly higher in RA than in OA. (II) Thrombin (0.1 to 100 units /ml) significantly enhanced growth of the synovial fibroblast-like cells (FC). When SFC were cultured with thrombin (100 units/ml) or MCT (10 ng/ml), concentrations of IL-8 and IL-6 in the culture medium were significantly increased compared with the control, while the concentrations of IL-1□, IL-1□ and TNF-□ were not detectable in all the culture mediums. (III) Cathepsin B-like protease purified from both the SF and synovial tissues ofRA patients was mainly composed of cathepsin B, and the cathepsin B purified from SF samples ofRA patients was capable of activating prourokinase to urokinase in vitro.

为了阐明蛋白酶在RA和OA中的病理生理意义，我们研究了RA和OA患者滑液和滑膜组织中哪些蛋白酶被特异性激活，以及丝氨酸蛋白酶对滑膜细胞释放炎性细胞因子的影响。(I)当蛋白酶底物与SF上清液孵育时，凝血酶最优先水解，而胰蛋白酶底物和组织蛋白酶B底物在一定程度上被水解。对于SF细胞提取物，组织蛋白酶B底物最优先水解，而其他底物几乎不水解。在滑膜组织提取物的情况下，组织蛋白酶B底物最优先水解，而胰蛋白酶底物在一定程度上水解。蛋白酶活性在一般显着高于RA比OA。(II)凝血酶（0.1至100单位/ml）显着增强滑膜成纤维细胞样细胞（FC）的生长。当SFC与凝血酶（100 units/ml）或MCT（10 ng/ml）共同培养时，培养液中IL-8和IL-6的浓度较对照组显著升高，而IL-1□、IL-1□和TNF-□在所有培养液中均未检测到。(III)从RA患者滑液和滑液中分离纯化的类组织蛋白酶B主要由组织蛋白酶B组成，且从RA患者滑液中分离纯化的组织蛋白酶B在体外可激活尿激酶原为尿激酶。