Clegg P D, Carter S D
Department of Veterinary Clinical Science and Animal Husbandry, University Veterinary Teaching Hospital, University of Liverpool, Neston, S. Wirral, UK.
Equine Vet J. 1999 Jul;31(4):324-30. doi: 10.1111/j.2042-3306.1999.tb03825.x.
A study was performed to identify the activation status of the gelatinase MMPs, MMP-2 and -9, in both normal and diseased equine articular tissues. In addition, the production and activation status of equine MMP-2 and -9 by equine articular cells and tissues in response to increasing IL-1beta concentrations was assessed. The study was performed to test the hypothesis that activation of MMPs is a fundamental step in the pathogenesis of joint diseases; and that this activation is mediated by the cytokine IL-1. Using purified equine MMP-2 and -9, the molecular weights of the zymogen and activated form of equine MMP-2 and -9 were identified by a combination of gelatin zymography and a gelatin degradation assay using aminophenylmercuric acetate as a chemical activator of the molecules. Normal equine articular tissues (cartilage and synovial membrane) maintained in short-term tissue culture produced MMP-2 zymogen alone, while similar tissues obtained from a variety of pathological conditions produce both zymogen and active MMP-2, as well as MMP-9 monomer and dimer. Activated MMP-9 was an inconsistent finding. Normal equine synovial fibroblasts in monolayer culture produced zymogen MMP-2 alone under basal conditions. A mild increase in active and zymogen MMP-2 levels occurred with IL-1beta treatment. Equine synovial membrane explants demonstrated a dose-dependent increase in active and zymogen MMP-2 and MMP-9 levels following IL-1beta treatment. Monolayer chondrocyte cell cultures demonstrated a dose-dependent mild increase in active and zymogen MMP-2 following IL-1beta treatment. Explant cartilage cultures demonstrated a dose-dependent mild increase in zymogen MMP-2 alone following IL-1beta treatment. This study supports the hypothesis that activation of MMPs is occurring in joint disease, and that in vitro stimulation of equine articular cells and tissues causes not only an increase in MMP production, but also an increase in amount of activated enzyme released. Further research is required to investigate the role of MMP activation in joint diseases, and to investigate the potential use of therapeutic agents, which inhibit MMP activation, in the treatment and prevention of joint diseases.
开展了一项研究,以确定明胶酶基质金属蛋白酶(MMP)-2和-9在正常和患病马关节组织中的激活状态。此外,还评估了马关节细胞和组织在白细胞介素(IL)-1β浓度增加时MMP-2和-9的产生及激活状态。该研究旨在验证以下假设:MMP激活是关节疾病发病机制中的一个基本步骤;且这种激活由细胞因子IL-1介导。使用纯化的马MMP-2和-9,通过明胶酶谱法和使用乙酸氨基苯汞作为分子化学激活剂的明胶降解试验相结合,确定了马MMP-2和-9的酶原及激活形式的分子量。短期组织培养中的正常马关节组织(软骨和滑膜)仅产生MMP-2酶原,而从各种病理状况下获得的类似组织则同时产生酶原和活性MMP-2,以及MMP-9单体和二聚体。激活的MMP-9的出现情况并不一致。单层培养的正常马滑膜成纤维细胞在基础条件下仅产生MMP-2酶原。IL-1β处理后,活性和酶原形式的MMP-2水平略有增加。马滑膜外植体在IL-1β处理后,活性和酶原形式的MMP-2及MMP-9水平呈剂量依赖性增加。单层软骨细胞培养物在IL-1β处理后,活性和酶原形式的MMP-2呈剂量依赖性轻度增加。外植体软骨培养物在IL-1β处理后,仅酶原形式的MMP-2呈剂量依赖性轻度增加。本研究支持以下假设:关节疾病中发生了MMP激活,并且体外刺激马关节细胞和组织不仅会导致MMP产生增加,还会导致释放的活性酶量增加。需要进一步研究来调查MMP激活在关节疾病中的作用,并研究抑制MMP激活的治疗药物在关节疾病治疗和预防中的潜在用途。