ADAMTS-9 is synergistically induced by interleukin-1β and tumor necrosis factor α in OUMS-27 chondrosarcoma cells and in human chondrocytes
ADAMTS-9 is synergistically induced by interleukin-1β and tumor necrosis factor α in OUMS-27 chondrosarcoma cells and in human chondrocytes
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DOI：
10.1002/art.21010
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发表时间：
2005-05-01
影响因子：
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通讯作者：
Ninomiya, Y
中科院分区：
文献类型：
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作者：
Demircan, K;Hirohata, S;Ninomiya, Y
objective.比较白细胞介素-1 β（IL-1 β）和肿瘤坏死因子α（TNF α）对软骨细胞样OUMS-27细胞和人软骨细胞中聚集蛋白聚糖酶（ADAMTS-1、ADAMTS-4、ADAMTS-5、ADAMTS-8、ADAMTS-9和ADAMTS-15）的诱导作用，并确定最敏感的聚集蛋白聚糖酶基因的诱导机制。将OUMS-27细胞用不同浓度的IL-1 β和/或TNF α刺激不同的时间段。还用IL-1 β和/或TNF α刺激从骨关节炎关节获得的人软骨细胞和人皮肤成纤维细胞。提取总RNA，逆转录，并通过定量实时聚合酶链反应和北方印迹分析。Western blotting检测ADAMTS-9蛋白的表达，探讨MAPK信号通路在IL-1 β刺激OUMS-27细胞中ADAMTS-9诱导表达中的作用。IL-1 β增加ADAMTS4、ADAMTS5和ADAMTS9的信使RNA（mRNA）水平，但不增加ADAMTS1和ADAMTS8。ADAMTS9 mRNA的增加倍数大于其他聚集蛋白聚糖酶基因的mRNA。在软骨细胞中，IL-1 β刺激引起的ADAMTS9 mRNA的增加大于成纤维细胞。IL-1 β和TNF α的组合具有协同效应，导致ADAMTS9 mRNA水平显著升高。ADAMTS-9蛋白在IL-1 β刺激的OUMS-27细胞中也被诱导。MAPK抑制剂SB203580和PD98059可降低ADAMTS9在OUMS-27细胞中的表达。ADAMTS9是一种IL-1 β和TNF α诱导型基因，似乎比其他聚集蛋白聚糖酶基因对这些促炎细胞因子更敏感。此外，这些细胞因子对ADAMTS9具有协同作用。结合已知ADAMTS-9蛋白水解降解聚集蛋白聚糖的能力及其切割其他软骨分子的潜力，数据表明ADAMTS-9可能在关节炎中具有病理作用。
Objective. To compare induction of the aggrecanases (ADAMTS-1, ADAMTS-4, ADAMTS-5, ADAMTS-8, ADAMTS-9, and ADAMTS-15) by interleukin-1 beta (IL-1 beta) and tumor necrosis factor a (TNF alpha) in chondrocyte-like OUMS-27 cells and human chondrocytes, and to determine the mechanism of induction of the most responsive aggrecanase gene.Methods. OUMS-27 cells were stimulated for different periods of time and with various concentrations of IL-1 beta and/or TNFa. Human chondrocytes obtained from osteoarthritic joints and human skin fibroblasts were also stimulated with IL-1 beta and/or TNF alpha. Total RNA was extracted, reverse transcribed, and analyzed by quantitative real-time polymerase chain reaction and Northern blotting. ADAMTS-9 protein was examined by Western blotting, and the role of the MAPK signaling pathway for ADAMTS9 induction in IL-1 beta-stimulated OUMS-27 cells was investigated.Results. IL-1 beta increased messenger RNA (mRNA) levels of ADAMTS4, ADAMTS5, and ADAMTS9 but not ADAMTS1 and ADAMTS8. The fold increase for ADAMTS9 mRNA was greater than that for mRNA of the other aggrecanase genes. The increase of ADAMTS9 mRNA by IL-1 beta stimulation was greater in chondrocytes than in fibroblasts. The combination of IL-1 beta and TNF alpha had a synergistic effect, resulting in a considerable elevation in the level of ADAMTS9 mRNA. ADAMTS-9 protein was also induced in IL-1 beta-stimulated OUMS-27 cells. The MAPK inhibitors SB203580 and PD98059 decreased ADAMTS9 upregulation in OUMS-27 cells.Conclusion. ADAMTS9 is an IL-1 beta- and TNF alpha-inducible gene that appears to be more responsive to these proinflammatory cytokines than are other aggrecanase genes. Furthermore, these cytokines had a synergistic effect on ADAMTS9. Together with the known ability of ADAMTS-9 to proteolytically degrade aggrecan and its potential to cleave other cartilage molecules, the data suggest that ADAMTS-9 may have a pathologic role in arthritis.