TNF-α increases expression of IL-6 and ICAM-1 genes through activation of NF-κB in osteoblast-like ROS17/2.8 cells
TNF-α increases expression of IL-6 and ICAM-1 genes through activation of NF-κB in osteoblast-like ROS17/2.8 cells
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DOI：
10.1359/jbmr.1998.13.8.1290
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发表时间：
1998-08-01
影响因子：
6.2
通讯作者：
Seo, H
中科院分区：
文献类型：
--
作者：
Kurokouchi, K;Kambe, F;Seo, H
肿瘤坏死因子-α（TNF-α）在炎性疾病如类风湿性关节炎和绝经后骨质疏松症中起关键作用。在各种组织中，TNF-α作用由转录因子核因子-κ B（NF-κ B）介导。然而，对于TNF-α如何在成骨细胞中发挥作用知之甚少，因此，我们检测了TNF-α对大鼠成骨样骨肉瘤细胞中NF-κ B活化的影响（ROS 17/2.8），电泳迁移率变动分析显示TNF-α早在15分钟就诱导了p50-p65异源二聚体NF-κ B的活化，随后持续活化48 h。当使用将NF-κ B与抑制性蛋白I κ B解离的去污剂检测细胞溶质中NF-κ B B的结合活性时，其在最初30分钟期间降低，然后增加至未刺激的水平。北方印迹分析显示，TNF-α作用6 h后，p50的前体p105 mRNA水平显著增加，而p65 mRNA水平在最初1 h内逐渐增加。两种mRNA水平的显著增加持续到TNF-α后24小时。这些结果表明，TNF-α对NF-κ B的快速激活主要是由于预先存在于胞质溶胶中的NF-κ B的核转位，并且随后p50和p65表达的增加可能导致TNF-α刺激期间NF-κ B的持续激活，TNF-α还增加白细胞介素-6（IL-6）和细胞间粘附分子-1（ICAM-1）的mRNA水平。抗氧化剂N-乙酰-L-半胱氨酸显著减弱了这些mRNA的TNF-α依赖性增加，同时降低了TNF-α对NF-κ B的激活，表明NF-κ B介导了IL-6和ICAM-1的TNF-α依赖性表达。在ROS 17/2.8细胞中。这些结果表明，TNF-α激活NF-κ B可能在成骨细胞产生细胞因子和细胞粘附分子中起重要作用，导致骨吸收和炎症的促进。
Tumor necrosis factor-alpha (TNF-alpha) plays a key role in inflammatory diseases such as rheumatoid arthritis and in postmenopausal osteoporosis, In various tissues, TNF-alpha action is mediated by a transcription factor, nuclear factor-kappa B (NF-kappa B). However, little is known about how TNF-alpha exerts its action in osteoblasts, We thus examined the effect of TNF-alpha on the activation of NF-kappa B in rat osteoblast-like osteosarcoma cells (ROS17/2.8), Electrophoretic mobility shift assay revealed that the activation of the p50-p65 heterodimer NF-kappa B was induced by TNF-alpha as early as 15 minutes followed by a persistent activation for 48 h. When the binding activity of NF-kappa B in cytosol was examined using detergents that dissociate NF-kappa B from an inhibitory protein I kappa B, it decreased during the initial 30 minutes 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. Significant increase in both mRNA levels continued until 24 h after TNF-alpha. These results suggest that the rapid activation of NF-kappa B by TNF-alpha is mainly due to the nuclear translocation of NF-kappa B pre-existing in cytosol, and that the subsequent increase in the expression of p50 and p65 may result in the persistent activation of NF-KB during TNF-alpha stimulation, TNF-alpha also increased the mRNA levels of interleukin-6 (IL-6) and intercellular adhesion molecule-1 (ICAM-1). An antioxidant, N-acetyl-L-cysteine, significantly attenuated the TNF-alpha-dependent increase in these mRNAs, and simultaneously reduced the activation of NF-kappa B by TNF-alpha, indicating that NF-kappa B mediates the TNF-alpha-dependent expression of IL-6 and ICAM-1. in ROS17/2,8 cells. These results suggest that the activation of NF-kappa B by TNF-alpha may play an important role In the production of cytokines and cell adhesion molecules from osteoblasts, leading to the promotion of bone resorption and inflammation.