WNT5A is induced by inflammatory mediators in bone marrow stromal cells and regulates cytokine and chemokine production
WNT5A is induced by inflammatory mediators in bone marrow stromal cells and regulates cytokine and chemokine production
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DOI：
10.1002/jbmr.1488
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发表时间：
2012-03-01
影响因子：
6.2
通讯作者：
Hofbauer, Lorenz C.
中科院分区：
文献类型：
--
作者：
Rauner, Martina;Stein, Nicola;Hofbauer, Lorenz C.
WNT5A最近被认为与炎症过程有关，但它作为骨髓基质细胞(BMSC)来源的介质在关节炎关节炎症中的作用尚不清楚。在这里，我们研究了炎症刺激是否诱导BMSC中的Wnt5A来控制炎症反应。用脂多糖(LPS)或肿瘤坏死因子α(TNF-α)刺激人骨髓间充质干细胞(BMSC)，并检测类风湿关节炎(RA)患者滑膜细胞和组织中WNT5A水平。利用Affymetrix基因芯片对Wnt5A处理的小鼠成骨细胞进行微阵列分析。用定量聚合酶链式反应、酶联免疫吸附试验和Luminex技术证实Wnt5A或Wnt5A基因敲除对骨髓间充质干细胞细胞因子/趋化因子表达的调节作用。使用特定的抑制剂确定了相关的信号通路。用荧光印迹系统检测MACS纯化的T淋巴细胞和单核细胞的迁移情况。BMSC经内毒素或肿瘤坏死因子-α刺激后，WNT5A的表达增加3倍。RA患者滑膜成纤维细胞中Wnt5A的表达是对照细胞的两倍，且在RA患者和hTNFtg小鼠的滑膜组织中Wnt5A的表达被高度诱导。Wnt5A处理的成骨细胞的微阵列分析确定了细胞因子和趋化因子为靶点。Wnt5A可诱导骨髓间充质干细胞产生IL-1β、IL-6、CCL2、CCL5、CXCL1和CXCL5，并依赖于核因子-β、丝裂原活化蛋白(MAPK)和Akt信号通路的激活。相应地，Wnt5A基因的敲除显著减少了基础和内毒素诱导的细胞因子/趋化因子的产生。单核细胞和T细胞向Wnt5A处理的BMSC上清液迁移分别增加25%和20%。这项研究强调了骨髓间充质干细胞来源的WNT5A在调节炎症过程中的关键作用，并提示它参与了RA的发病机制。(C)2012年美国骨与矿物研究学会。
WNT5A has recently been implicated in inflammatory processes, but its role as a bone marrow stromal cell (BMSC)derived mediator of joint inflammation in arthritis is unclear. Here, we investigated whether inflammatory stimuli induce WNT5A in BMSC to control inflammatory responses. WNT5A levels were determined in human BMSC after stimulation with lipopolysaccharide (LPS) or tumor necrosis factor a (TNF-a,) and in synovial cells and tissue of patients with rheumatoid arthritis (RA) and human TNF-a transgenic (hTNFtg) mice. A microarray analysis of WNT5A-treated murine osteoblasts was performed using Affymetrix gene chips. The regulation of cytokine/chemokine expression was confirmed by qPCR, ELISA, and Luminex technology in BMSC after stimulation with WNT5A or WNT5A knockdown. Relevant signaling pathways were identified using specific inhibitors. Migration of MACS-purified T lymphocytes and monocytes was assessed using the FluoroBlok system. WNT5A expression was increased threefold in BMSC after stimulation with LPS or TNF-a. Synovial fibroblasts from patients with RA showed a twofold increase of WNT5A expression compared with control cells, and its expression was highly induced in the synovial tissue of patients with RA and hTNFtg mice. Microarray analysis of WNT5A-treated osteoblasts identified cytokines and chemokines as targets. The induction of IL-1 beta, IL-6, CCL2, CCL5, CXCL1, and CXCL5 by WNT5A was confirmed in BMSC and depended on the activation of the NF-?B, mitogen-activated protein (MAPK), and Akt pathways. Accordingly, knockdown of WNT5A markedly reduced the basal and LPS-induced cytokine/chemokine production. Finally, migration of monocytes and T cells toward the supernatant of WNT5A-treated BMSC was increased by 25% and 20%, respectively. This study underlines the critical role of BMSC-derived WNT5A in the regulation of inflammatory processes and suggests its participation in the pathogenesis of RA. (c) 2012 American Society for Bone and Mineral Research.