Characterisation of equine matrix metalloproteinase 2 and 9; and identification of the cellular sources of these enzymes in joints
Characterisation of equine matrix metalloproteinase 2 and 9; and identification of the cellular sources of these enzymes in joints
复制标题
DOI：
10.1111/j.2042-3306.1997.tb03136.x
复制
发表时间：
1997-09-01
影响因子：
2.2
通讯作者：
Carter, SD
中科院分区：
文献类型：
--
作者：
Clegg, PD;Burke, RM;Carter, SD
通过组织培养方法和明胶酶谱法分析细胞上清液，研究了常驻关节细胞和浸润炎性细胞的明胶酶基质金属蛋白酶（MMP）的细胞产生。在短期培养中的外周血中性粒细胞产生MMP-9，在培养中的外周血单核细胞也是如此。分离的关节软骨细胞在单层培养产生MMP-2和MMP-9，虽然关节软骨保持作为外植体培养产生MMP-2单独。单层培养中生长的滑膜成纤维细胞仅产生MMP-2，尽管外植体培养中的滑膜产生MMP-2和MMP-2的活性形式。血液多形中性粒细胞的裂解产生大量MMP-9，但血液单核细胞、滑膜成纤维细胞和关节软骨细胞的裂解产生很少的酶，这表明，与其他细胞类型不同，多形中性粒细胞在细胞内储存MMP，用明胶-琼脂糖亲和层析法从滑膜成纤维细胞培养上清中分离纯化出马MMP-2，从多形中性粒细胞培养上清中分离纯化出马MMP-9。侵袭细胞和驻留关节细胞可以产生能够消化关节软骨基质的某些组分分子的酶的证明表明，这些酶的治疗靶向可能是预防骨关节炎中软骨破坏的有效主张。
The cellular production by resident articular cells and infiltrating inflammatory cells of the gelatinase matrix metalloproteinases (MMP) was investigated by tissue culture methods and analysis of cell supernatants by gelatin zymography. Peripheral blood neutrophils in short term culture produced MMP-9, as did peripheral blood monocytes in culture. Isolated articular chondrocytes in monolayer culture produced both MMP-2 and MMP-9, although articular cartilage maintained as explant culture produced MMP-2 alone. Synovial fibroblasts grown in monolayer culture produced MMP-2 alone, although synovial membrane in explant culture produced both MMP-2 and the active form of MMP-2, Lysis of blood polymorph neutrophils produced large quantities of MMP-9, but lysis of blood monocytes, synovial fibroblasts and articular chondrocytes produced little enzyme indicating that, unlike the other cell types, polymorph neutrophils store MMPs intracellularly, Equine MMP-2 was purified from synovial fibroblast cell culture supernatant, and equine MMP-9 from polymorph neutrophil cell culture supernatant, by gelatin-sepharose affinity chromatography, The 2 enzymes were identified from their molecular weights and by their respective N-terminal amino acid sequences which showed homology with the enzymes from other species. The demonstration that invasive cells and resident articular cells can produce enzymes which are capable of digestion of certain component molecules of the articular cartilage matrix, shows that therapeutic targeting of these enzymes could be a valid proposition in the prevention of cartilage destruction in osteoarthritis.