TLR-3 enhances osteoclastogenesis through upregulation of RANKL expression from fibroblast-like synoviocytes in patients with rheumatoid arthritis
TLR-3 enhances osteoclastogenesis through upregulation of RANKL expression from fibroblast-like synoviocytes in patients with rheumatoid arthritis
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DOI：
10.1016/j.imlet.2009.02.006
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发表时间：
2009-05-14
影响因子：
4.4
通讯作者：
Kim, Ho-Youn
中科院分区：
文献类型：
--
作者：
Kim, Kyoung-Woon;Cho, Mi-La;Kim, Ho-Youn
为探讨Toll样受体-3(Toll-like Receptor-3，TLR3)对类风湿关节炎(RA)成纤维样滑膜细胞(FLS)破骨活性的调节作用，采用RT-PCR方法检测TLR3激活后RA-FLS中核因子kappa B受体激活剂B配体(RANKL)基因和蛋白的表达。实时荧光定量聚合酶链式反应、蛋白印迹分析和免疫组织化学检测。人单核细胞与经TLR3配体多聚核苷-多核胞苷酸预刺激的RA-FLS共培养，然后进行抗酒石酸酸性磷酸酶(TRAP)染色。用RT-PCR和实时定量聚合酶链式反应检测破骨细胞的其他标志物。TLR3和RANKL在RA滑膜中的表达明显高于在骨关节炎(OA)滑膜中的表达。TLR3激活可诱导RA-FLS表达RANKL，但对OA-FLS和正常皮肤成纤维细胞无明显影响。TLR3激活也可诱导RA-FLS产生IL-1β，但对IL-17和TNF-α的产生无影响。抑制IL-1β可逆转TLR3诱导的RANKL表达上调。人单核细胞与TLR3激活的RA-FLS或TLR3配体刺激的人单核细胞共培养后，TRAP、RANK、组织蛋白酶K、降钙素受体和基质金属蛋白酶-9的表达增加，反映了单核细胞向破骨细胞的分化。TLR3直接刺激人单核细胞，促进破骨细胞分化。TLR3间接诱导RA-FLS中RANKL的表达，RANKL的表达促进RA滑膜破骨细胞的分化。靶向TLR3通路可能是预防RA炎性骨破坏的一种有前途的方法。(C)2009爱思唯尔B.V.保留所有权利。
This study was undertaken to determine the effect of toll-like receptor-3 (TLR3) on the regulation of osteoclastogenic activity in rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLS).The expression of receptor activator of nuclear factor kappa B ligand (RANKL) mRNA and protein in RA-FLS after TLR3 activation was determined using RT-PCR. real-time PCR, western blot analysis, and immunohistochemistry. Human monocytes were cocultured with RA-FLS that had been prestimulated by the TLR3 ligand polyriboinosinic-polyribocytidylic acid and then stained for tartrate-resistant acid phosphatase (TRAP) activity. Other markers of osteoclasts were measured using RT-PCR and real-time PCR.The expression of TLR3 and RANKL was much higher in the RA synovium than in the osteoarthritis (OA) synovium. TLR3 activation induced RANKL expression in RA-FLS, but not in OA-FLS or in normal skin fibroblasts. TLR3 activation also induced the production of IL-1 beta but had no effect on IL-17 or TNF-alpha production in RA-FLS. Inhibition of IL-1 beta reversed the TLR3-induced upregulation of RANKL expression. Coculture of human monocytes with TLR3-activated RA-FLS or TLR3 ligand-stimulated human monocytes increased the expression of TRAP, RANK, cathepsin K, calcitonin receptor, and MMP-9, reflecting the differentiation of monocytes into osteoclasts.Our results suggest that TLR3 promotes osteoclastogenesis in the RA synovium both directly and indirectly. TLR3 stimulates human monocytes directly to promote osteoclast differentiation. TLR3 induces RANKL expression indirectly in RA-FLS, and the expression of RANKL promotes the differentiation of osteoclasts in the RA synovium. Targeting the TLR3 pathway may be a promising approach to preventing inflammatory bone destruction in RA. (c) 2009 Elsevier B.V. All rights reserved.