The effects of degenerative products from matrix on syanovial cells and chondrocytes metabolism

采集54例骨性关节炎患者膝关节滑液标本。对34例膝骨关节炎患者进行了X线片和核磁共振(MR)成像评估，以确定疾病的分期。TIMP-1的含量与基质金属蛋白酶-1、-3的含量呈正相关，两者也呈正相关，证实了早期的研究结果。DELTAdi-C6S/Deltadi-C4S比值与MMP3呈负相关。DELTAdi-C6s与KS表位相关，与846表位(后者也与ddi-C4S)相关程度较小。TIMP-1与846表位相关，TIMP-2与CPII相关。在晚期OA的SF中，DELTAdi-C6S、DELTAdi-C4S、846表位和CPII的浓度显著降低。TIMP的产生似乎分别与特定软骨分子的合成有关。软骨基质结构成分含量的减少表明存在可测量的软骨la…的丢失。如前所述，在疾病中有更多的TE。因此，我们检测了肿瘤坏死因子-α对大鼠成骨样骨肉瘤细胞(ROS17/2.8)中核因子-kappaB活化的影响。凝胶迁移率改变分析显示，p5O-p65异质结合体NF-kappaB在15 mM时被TNF-α诱导激活，随后持续激活48h，胞浆中的结合活性在开始的30min内下降，然后升高到未刺激的水平。Northern印迹分析显示，在肿瘤坏死因子-α作用后6h，p50的前体p105的mRNA水平显著升高，在最初的1h内，p65的mRNA水平逐渐升高，抗氧化剂显著减弱了这些mRNAs对肿瘤坏死因子-α的依赖性增加，同时也降低了由肿瘤坏死因子-α引起的核因子-kappaB的活化。这些结果提示，肿瘤坏死因子-α激活的核因子-kappaB可能在成骨细胞产生细胞因子的过程中起重要作用，从而促进骨吸收和炎症。较少

Synovial fluid samples from the knees of 54 patients with OA were collected. Radiographic and magnetic resonance (MR) imaging evaluations were performed on 34 of the OA patientsi knees to classify the stage of the disease. The content of TIMP-1 was directly correlated with those of MMP-1 and -3, which were also correlated with each other, confirming earlier results. There was an inverse correlation between the ratio DELTAdi-C6S/deltadi-C4S and MMP-3. DELTAdi-C6S was correlated with the KS epitope, and to a lesser degree with the 846 epitope (latter also with Ddi-C4S). TIMP-1 correlated with the 846 epitope, TIMP-2 correlated with the CPII.There were significantly lower concentrations of DELTAdi-C6S, DELTAdi-C4S, the 846 epitope and CPII in SF in late stage OA.The production of TIMPs appears to be individually linked to the synthesis of specific cartilage molecules. The reduction in content of cartilage matrix structural components suggest that there is a measurable loss of cartilage la … More te in the disease as suggested previously. We thus examined the effect of TNF-alpha on the activation of NF-kappaB in rat osteoblast-like osteosarcoma cells (ROS17/2.8) . Electrophoretic mobility shift assay revealed that the activation of the p5O-p65 heteroclimer NF-kappaB was induced by TNF-alpha as early as 15 mm followed by a persistent activation for 48 h. The binding activity of NF-kappaB in cytosol decreased during the initial 30 min and then increased to the unstimulated level. Northern blot analysis revealed a marked increase in the mRNA levels of p105, a precursor of p50, 6 h after TNF-alpha and a gradual increase in p65 mRNA levels during the initial 1 h. An antioxidant significantly attenuated the TNF-alpha-dependent increase in these mRNAs, and simultaneously reduced the activation of NF-kappaB by TNF-alpha. These results suggest that the activation of NF-kappaB by TNF-alpha may play an important role in the production of cytokines from osteoblasts, leading to the promotion of bone resorption and inflammation. Less