Rédini F, Lafuma C, Pujol J P, Robert L, Hornebeck W
Laboratoire de Biochimie du Tissu Conjonctif, CHU Côte de Nacre, Caen, France.
Biochem Biophys Res Commun. 1988 Sep 15;155(2):786-93. doi: 10.1016/s0006-291x(88)80564-3.
Human synoviocytes, rabbit articular chondrocytes and human skin fibroblasts in culture were examined for their ability to express elastase activity. Latent enzyme activity degrading insoluble elastin was detected in the culture media of the three cell types and was completely abolished by metal chelating agents. Triton X-100 cell extracts were found to degrade a synthetic elastase substrate, N Succinyl-(Ala)3p-nitroanilide (SANA). The SANA-degrading activity of cell extracts could be attributed to a metalloprotease for fibroblasts and synoviocytes (100%) and to a metalloprotease associated with a cysteine protease for chondrocytes (70 and 30% respectively). This SANA-degrading activity was partly due to the combined action of an endo and an exopeptidase. Tumor Necrosis Factor-alpha (TNF-alpha) and Interferon-gamma (IFN-gamma) significantly enhanced the elastin degrading activity present in the culture media of both synoviocytes and chondrocytes. Interleukin-1 beta significantly increased the secretion of elastase by chondrocytes. By contrast, Transforming Growth Factor-beta (TGF-beta) reduced by 80 per cent the secretion of elastinolytic activity by chondrocytes but had not effect on other cell types.
检测了培养的人滑膜细胞、兔关节软骨细胞和人皮肤成纤维细胞表达弹性蛋白酶活性的能力。在这三种细胞类型的培养基中均检测到了降解不溶性弹性蛋白的潜在酶活性,且金属螯合剂可使其完全丧失。发现Triton X-100细胞提取物可降解合成弹性蛋白酶底物N-琥珀酰-(丙氨酸)3-对硝基苯胺(SANA)。细胞提取物的SANA降解活性对于成纤维细胞和滑膜细胞可归因于一种金属蛋白酶(100%),对于软骨细胞则归因于一种与半胱氨酸蛋白酶相关的金属蛋白酶(分别为70%和30%)。这种SANA降解活性部分归因于一种内切肽酶和一种外切肽酶的联合作用。肿瘤坏死因子-α(TNF-α)和干扰素-γ(IFN-γ)显著增强了滑膜细胞和软骨细胞培养基中存在的弹性蛋白降解活性。白细胞介素-1β显著增加了软骨细胞弹性蛋白酶的分泌。相比之下,转化生长因子-β(TGF-β)使软骨细胞弹性溶解活性的分泌减少了80%,但对其他细胞类型没有影响。