Activation of NF-κB signalling and TNFα-expression in THP-1 macrophages by TiAlV- and polyethylene-wear particles
Activation of NF-κB signalling and TNFα-expression in THP-1 macrophages by TiAlV- and polyethylene-wear particles
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DOI：
10.1016/j.orthres.2005.02.017
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发表时间：
2005-11-01
影响因子：
2.8
通讯作者：
Rader, CP
中科院分区：
文献类型：
--
作者：
Baumann, B;Seufert, J;Rader, CP
磨损颗粒被认为会诱发假体周围炎症，从而导致假体周围骨质溶解。TNF α在该过程的发病机制中起关键作用。导致假体周围炎症发展的分子机制尚未确定，单核细胞对不同磨损颗粒的反应是TNF α表达上调。在本研究中，我们评价了聚乙烯和TiAl V颗粒对假体周围破骨细胞生成方面单核细胞中NF-κ B信号通路活化和TNF α生物合成和释放的影响。将THP-1单核细胞分化为巨噬细胞样细胞，并暴露于LPS解毒的聚乙烯和假体衍生的TiAlV颗粒。通过ELISA分析培养物上清液中的TNF α释放。通过电泳迁移率变动试验（EMSA）检查NF-κ B活化，并通过荧光素酶报告基因试验测定NF-κ B靶启动子活性（包括TNF α启动子的反式激活）。分化的THP-1巨噬细胞暴露于增加数量的颗粒0，60，180和360分钟。这两个，聚乙烯和TiAlV颗粒诱导一个显着的激活NF-κ B和TNF α启动子在180分钟。一个显着的TNF α释放后检测到360分钟暴露于聚乙烯和TiAlV颗粒的剂量依赖性方式。相比之下，LPS诱导NF-κ B和TNF α启动子的更大活化，并且强烈诱导TNF α分泌到上清液中。这些结果提供了证据，表明巨噬细胞中NF-κ B信号转导通路的诱导在引发和介导导致假体周围骨质溶解的炎症反应中起主要作用。(c)2005骨科研究学会。由爱思唯尔有限公司出版。保留所有权利。
Wear particles are believed to induce periprosthetic inflammation which contributes to periprosthetic osteolysis. TNF alpha plays a pivotal role in the pathogenesis of this process. The molecular mechanisms leading to the development of periprosthetic inflammation with upregulated TNF alpha expression in monocytic cells in response to different wear particles have yet to be defined. In this study we evaluated the effects of polyethylene- and TiAlV-particles on activation of NF-kappa B signalling pathways and TNF alpha biosynthesis and release in monocytic cells with respect to periprosthetic osteoclastogenesis. THP-1 monocytic cells were differentiated to macrophage-like cells and exposed to LPS-detoxified polyethylene and prosthesis-derived TiAlV-particles. TNF alpha release was analyzed in culture supernatant by ELISA. NF-kappa B activation was examined by electrophoretic mobility shift assay (EMSA), and NF-kappa B target promoter activities including transactivation of the TNF alpha promoter were determined by luciferase reporter gene assays. Differentiated THP-1 macrophages were exposed to increasing numbers of particles for 0, 60, 180 and 360 min. Both, polyethylene- and TiAlV-particles induced a significant activation of both NF-kappa B and TNF alpha promoters at 180 min. A significant TNF alpha release was detected after 360 min exposure to polyethylene- and TiAlV-particles in a dose dependent manner. In comparison, LPS induced a much greater activation of NF-kappa B and TNF alpha promoters, and TNF alpha secretion into the supernatant was strongly induced. These results provide evidence that induction of the NF-kappa B signal transduction pathway in macrophages plays a major role in initiating and mediating the inflammatory response leading to periprosthetic osteolysis. (c) 2005 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved.