Diphenylhydantoin Inhibits Osteoclast Differentiation and Function Through Suppression of NFATc1 Signaling
Diphenylhydantoin Inhibits Osteoclast Differentiation and Function Through Suppression of NFATc1 Signaling
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DOI：
10.1359/jbmr.090302
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发表时间：
2009-08-01
影响因子：
6.2
通讯作者：
Udagawa, Nobuyuki
中科院分区：
文献类型：
--
作者：
Koide, Masanori;Kinugawa, Saya;Udagawa, Nobuyuki
二苯海因(DPH)是一种广泛应用的抗惊厥药物。我们使用体内和体外检测系统检测了DPH对破骨细胞分化和功能的影响。过量表达可溶性RANKL(RANKL TG)的转基因小鼠表现出更多的破骨细胞性骨吸收。将DPH注射到RANKL转基因小鼠的头盖骨上覆的皮下组织中，可抑制头盖骨的增强吸收。在小鼠成骨细胞和骨髓细胞的共培养中，DPH抑制了脂多糖(LPS)诱导的破骨细胞的形成。DPH对体外培养的小鼠成骨细胞的生长和RANKL、骨保护素基因的表达均无影响。另一方面，DPh抑制RANKL诱导的小鼠骨髓来源的巨噬细胞(Phi S)和人外周血源性CD14(+)细胞的破骨细胞的形成。DPH可抑制小鼠颅骨器官培养中脂多糖诱导的骨吸收，并抑制牙本质片上培养的破骨细胞的凹坑形成活性。DPh可抑制RANKL诱导的T细胞钙振荡和活化T细胞核因子c1(NFATc1)和c-fos的表达，并抑制RANKL诱导的NFATc1的核定位和自身扩增。在成熟的破骨细胞中。骨髓基质细胞和破骨细胞都表达T型钙通道Cav3.2的基因，Cav3.2是DPH的靶标。短发夹状RNA阻断Cav3.2的表达可显著抑制RANKL诱导的破骨细胞分化。这些结果提示DPH通过抑制NFATc1信号通路抑制破骨细胞的分化和功能。局部应用DPH可能是预防牙周炎等局部炎症引起的骨丢失的一种治疗方法。《矿工杂志》2009；24：1469-1480。2009年3月16日在线发布；DOI：10.1359/JBMR.090302
Diphenylhydantoin (DPH) is widely used as an anticonvulsant drug. We examined the effects of DPH on osteoclast differentiation and function using in vivo and in vitro assay systems. Transgenic mice overexpressing a soluble form of RANKL (RANKL Tg) exhibited increased osteoclastic bone resorption. Injection of DPH into the subcutaneous tissue overlying calvaria of RANKL Tg mice suppressed the enhanced resorption in the calvaria. In co-cultures of mouse osteoblasts and bone marrow cells, DPH inhibited lipopolysaccharide (LPS)-induced osteoclast formation. DPH affected neither the mRNA expression of RANKL and osteoprotegerin nor the growth of mouse osteoblasts in culture. On the other hand, DPH inhibited the RANKL-induced formation of osteoclasts in cultures of mouse bone marrow-derived macrophages (BMM phi s) and of human peripheral blood-derived CD14(+) cells. DPH concealed LPS-induced bone resorption in mouse calvarial organ cultures and inhibited the pit-forming activity of mouse osteoclasts cultured on dentine slices. DPH suppressed the RANKL-induced calcium oscillation and expression of nuclear factor of activated T cells c1 (NFATc1) and c-fos in BMM phi s. Moreover, DPH inhibited the RANKL-induced nuclear localization and auto-amplification of NFATc1. in mature osteoclasts. Both BMM phi s and osteoclasts expressed mRNA of a T-type calcium channel, Cav3.2, a target of DPH. Blocking the expression of Cav3.2 by short hairpin RNAs significantly suppressed RANKL-induced osteoclast differentiation. These results suggest that DPH inhibits osteoclast differentiation and function through suppression of NFATc1 signaling. The topical application of DPH may be a therapeutic treatment to prevent bone loss induced by local inflammation such as periodontitis. J Bone Miner Res 2009;24:1469-1480. Published online on March 16, 2009; doi: 10.1359/JBMR.090302