Homandberg G A, Hui F
Department of Biochemistry, Rush Medical College at Rush-Presbyterian-St. Luke's Medical Center, Chicago, Illinois 60612-3864, USA.
Arch Biochem Biophys. 1996 Oct 15;334(2):325-31. doi: 10.1006/abbi.1996.0461.
Addition of fibronectin fragments to bovine articular cartilage explant cultures results in enhanced release of metalloproteinases and rapid cartilage proteoglycan (PG) degradation and loss. The chondrolysis begins with rapid PG degradation which markedly slows after 1 week. Preliminary observations suggest that catabolic cytokines mediate chondrolytic activities of the fibronectin fragments. The objectives of this work were to investigate the correlations between: (a) release of specific cytokines; (b) release of the metalloproteinase (MMP), stromelysin-1 (MMP-3); (c) release of the tissue inhibitor of MMPs, TIMP-1, and; (d) degradation and release of PG from cultured cartilage. We report that human articular cartilage cultured with an amino-terminal 29-kDa fragment (Fn-f) at 0.1 microM, released enhanced levels of TNF-alpha, IL-1beta, and IL-1alpha with peaks at Days 2, 3, and 9, respectively. MMP-3 release was elevated with a peak at Day 6 and a profile similar to that for the Fn-f-induced cartilage PG depletion. IL-6 release was enhanced within 2 days and continued at the same level throughout the culture period but this did not lead to enhanced release of TIMP-1, a known activity of IL-6. These data suggest that in the early chondrolytic events induced in cultured cartilage by Fn-f, enhanced MMP-3 release and maximal degradation and release of PG from cultured cartilage are kinetically associated with elevated release of the catabolic cytokines, TNF-alpha, IL-1beta, and IL-1alpha. Further, a later period of slowing PG loss and slowing MMP-3 release is associated with greatly slowed release of these cytokines, but prolonged release of IL-6. This model of cartilage damage may be useful for studies of the interplay between cytokines and the effects of combinations of cytokines on cartilage homeostasis.
向牛关节软骨外植体培养物中添加纤连蛋白片段会导致金属蛋白酶释放增加以及软骨蛋白聚糖(PG)快速降解和流失。软骨溶解始于PG的快速降解,1周后降解速度明显减慢。初步观察表明,分解代谢细胞因子介导纤连蛋白片段的软骨溶解活性。这项工作的目的是研究以下各项之间的相关性:(a)特定细胞因子的释放;(b)金属蛋白酶基质溶解素-1(MMP-3)的释放;(c)MMP组织抑制剂TIMP-1的释放;以及(d)培养软骨中PG的降解和释放。我们报告,用0.1微摩尔的氨基末端29 kDa片段(Fn-f)培养的人关节软骨,分别在第2天、第3天和第9天释放出水平升高的TNF-α、IL-1β和IL-1α,达到峰值。MMP-3的释放在第6天升高并达到峰值,其模式与Fn-f诱导的软骨PG消耗相似。IL-6的释放在2天内增强,并在整个培养期间保持在同一水平,但这并未导致TIMP-1释放增加,而TIMP-1释放增加是IL-6的已知活性。这些数据表明,在Fn-f诱导的培养软骨早期软骨溶解事件中,MMP-3释放增加以及培养软骨中PG的最大降解和释放与分解代谢细胞因子TNF-α、IL-1β和IL-1α的释放增加在动力学上相关。此外,后期PG流失减缓以及MMP-3释放减缓与这些细胞因子的释放大幅减缓有关,但IL-6释放持续时间延长。这种软骨损伤模型可能有助于研究细胞因子之间的相互作用以及细胞因子组合对软骨稳态的影响。