Differential induction and regulation of matrix metalloproteinases in osteoarthritic tissue and fluid synovial fibroblasts
Differential induction and regulation of matrix metalloproteinases in osteoarthritic tissue and fluid synovial fibroblasts
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DOI：
10.1016/j.joca.2004.02.005
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发表时间：
2004-05-01
影响因子：
7
通讯作者：
Dankbar, B
中科院分区：
文献类型：
--
作者：
Fuchs, S;Skwara, A;Dankbar, B
目的：研究滑膜液来源的成纤维细胞分泌基质金属蛋白酶（MMP）及其抑制剂（TIMP）的情况，并与组织来源的成纤维细胞进行比较。用肿瘤坏死因子（TNF）-α、白细胞介素（IL）-1 α、IL-1 β、IL-6以及TNF α和IL-1 β的组合。用细胞因子组合或骨关节炎滑液刺激成纤维细胞和软骨的共培养物。采用酶联免疫吸附法检测MMP-1、MMP-3、MMP-8、MMP-13、TIMP-1和TIMP-2的分泌。明胶酶谱和免疫印迹进行证明酶activity.Results：TNF α，IL-1 α，IL-1 β导致显着增加MMP-1和MMP-3的释放（高达4.2倍和547倍，分别）滑膜成纤维细胞，而分泌MMP-13诱导TNF α和IL-1 β的伴随管理。细胞内MMP-8的表达被细胞因子刺激，但释放所需的滑膜成纤维细胞与软骨的粘附。在整个研究中，与TSC培养物相比，在刺激的FSC中观察到显著更高水平的分泌的MMP。此外，MMP分泌的增加并不伴随着分泌的TIMP-1和TIMP-2的增加，导致酶和抑制剂levels.Conclusions之间的显着不平衡：结果提供了强有力的证据，滑膜来源的基质金属蛋白酶对软骨破坏OA的显着影响。在这种情况下，滑膜液中存在的成纤维细胞似乎发挥了突出的作用。(C)2004年国际骨关节炎研究学会。由爱思唯尔有限公司出版。保留所有权利。
Objectives: To investigate the secretion profiles of matrix metalloproteinases (MMP) and their inhibitors (TIMP) in synovial fluid-derived fibroblasts and to compare them with those of tissue-derived fibroblasts.Methods: Fibroblast cultures established from synovial tissues (TSC) and fluids (FSC) of the same OA patients were stimulated with tumor necrosis factor(TNF)-alpha, interleukin(IL)-1alpha, IL-1beta, IL-6 and a combination of TNFalpha and IL-1beta. Cocultures of fibroblasts and cartilage were stimulated either with the cytokine combination or with osteoarthritic synovial fluid. Secretion of MMP-1, MMP-3, MMP-8, MMP-13, TIMP-1, and TIMP-2 was measured by enzyme-linked immunosorbent assay. Gelatin zymography and immunoblotting were performed to demonstrate enzyme activity.Results: TNFalpha, IL-1alpha, and IL-1beta led to marked increases in MMP-1 and MMP-3 release (up to 4.2-fold and 547-fold, respectively) by synovial fibroblasts, whereas secretion of MMP-13 was induced by concomitant administration of TNFalpha and IL-1beta. Expression of intracellular MMP-8 was stimulated by cytokines, but adhesion of synovial fibroblasts to cartilage was required for the release. Throughout the study, significantly higher levels of secreted MMPs were observed in stimulated FSC compared to TSC cultures. Furthermore, increases in MMP secretion were not accompanied by increases in secreted TIMP-1 and TIMP-2, resulting in marked imbalances between enzyme and inhibitor levels.Conclusions: The results provide strong evidence for a significant impact of synovial-derived MMPs on cartilage destruction in OA. In this context, fibroblasts present in the synovial fluid appeared to play an outstanding role. (C) 2004 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved.