Iguratimod (T-614) suppresses RANKL-induced osteoclast differentiation and migration in RAW264.7 cells via NF-κB and MAPK pathways
Iguratimod (T-614) suppresses RANKL-induced osteoclast differentiation and migration in RAW264.7 cells via NF-κB and MAPK pathways
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Iguatimod (T-614) 通过 NF-kappa B 和 MAPK 途径抑制 RANKL 诱导的 RAW264.7 细胞中的破骨细胞分化和迁移
DOI：
10.1016/j.intimp.2016.03.038
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发表时间：
2016-06-01
影响因子：
5.6
通讯作者：
Zhang, Miaojia
中科院分区：
文献类型：
--
作者：
Gan, Ke;Yang, Leilei;Zhang, Miaojia
简介：艾拉莫德（Iguratimod，T-614）是一种高效、安全的治疗类风湿关节炎的新型药物，在中国和日本已被证实具有良好的抗炎作用。方法：采用TRAP染色法、transwell迁移实验和成骨细胞盘法分别观察艾拉莫德对破骨细胞分化、迁移和骨吸收的影响。RT-PCR检测破骨细胞相关基因、趋化因子和转录因子的相对表达，Western blotting检测破骨细胞相关信号传导通路。结果：艾拉莫德能明显抑制RANKL诱导的RAW 264. 7细胞的破骨细胞分化、迁移和骨吸收，并呈剂量依赖性。RANKL刺激后RAW264.7细胞TRAP、CTSK和CTR等骨诱导相关基因表达增加，而Iguratimod则明显抑制其表达。Iguratimod可剂量依赖性地抑制RANKL诱导的趋化因子CCL 7、CCL 4和CCL 12的表达以及骨形成相关转录因子c-Fos、c-Jun和NFATc 1的表达。Western blotting结果显示，艾拉莫德可抑制RANKL诱导的RAW 264. 7破骨细胞生成过程中MAPK和NF-κ B通路的激活。结论：艾拉莫德对破骨细胞的形成和功能有直接的抑制作用，这与以往的研究结果不同，提示艾拉莫德对RA有独特的治疗作用，尤其是在预防骨破坏方面。(C)© 2016 Elsevier B. V.版权所有。
Introduction: Iguratimod (T-614) has been confirmed as a highly efficacious and safe novel disease-modifying anti-rheumatic drug (DMARD) for rheumatoid arthritis therapy in China and Japan due to its potent anti inflammation effect. Here, we investigate the effects of Iguratimod on osteoclast differentiation, migration and function.Methods: The effect of Iguratimod on osteoclastogenesis, migration and bone resorption were assessed by TRAP staining, transwell migration assay and osteologic discs, respectively. Relative expressions of osteoclastic related genes, chemokines and transcription factors were assessed by reverse transcription polymerase chain reaction (RT-PCR) and signaling pathways were analyzed by western blotting.Results: Iguratimod significantly inhibits osteoclast differentiation, migration and bone resorption in RANKL-induced RAW264.7 cell in a dose-dependent manner. The expressions of osteoclastic related genes including TRAP, CTSK and CTR were increased in RAW264.7 cell upon RANKL stimulation but were obviously suppressed in the presence of Iguratimod. RANKL induced the expression of chemokines including CCL7, CCL4 and CCL12 and osteoclastic related transcription factors of c-Fos, c-Jun and NFATc1 could be significantly inhibited by Iguratimod in a dose dependent manner. Western blotting indicated Iguratimod could suppress the activation of MAPKs and NF-kappa B pathway in RANKL induced osteoclastogenesis in RAW264.7.Conclusions: These findings revealed a directly inhibitory role of Iguratimod on osteoclast formation and function, which is distinct from previous report, suggesting Iguratimod provide a unique therapeutic strategy for RA and especially in light of preventing bone destruction. (C) 2016 Elsevier B.V. All rights reserved.