Oral malodorous compound induces osteoclast differentiation without receptor activator of nuclear factor κB ligand.
Oral malodorous compound induces osteoclast differentiation without receptor activator of nuclear factor κB ligand.
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DOI：
10.1902/jop.2010.100116
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发表时间：
2010-11
影响因子：
4.3
通讯作者：
Hisataka Ii;T. Imai;K. Yaegaki;K. Irie;D. Ekuni;M. Morita
中科院分区：
文献类型：
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作者：
Hisataka Ii;T. Imai;K. Yaegaki;K. Irie;D. Ekuni;M. Morita
背景硫化氢（H（2）S）是口臭的主要成分，是牙周炎的致病因素之一。我们最近报道H（2）S可能会引起大鼠牙槽骨的病理变化。本研究的目的是确定H（2）S对破骨细胞分化的影响。方法将小鼠巨噬细胞RAW 264置于不含核因子κB配体（核因子κB配体的受体激活剂）的培养液中，于37°C、5%CO2和空气中培养24 h，然后加入0.05、0.5或5 ng/ml硫化氢，培养4 d。对照组接受不含硫化氢的CO（2）-空气混合物。通过计数抗酒石酸酸性磷酸酶（TRAP）阳性细胞来评价细胞分化。Western blotting检测细胞外信号调节激酶1/2（ERK 1/2）和丝裂原活化蛋白激酶p38磷酸化水平。骨吸收活性用磷酸钙吸收法测定。结果0.05ng/ml硫化氢组TRAP阳性细胞数明显多于其他浓度组（P <0.001）;破骨细胞的特异性标志物组织蛋白酶K蛋白在H（2）S诱导的多核细胞中表达。在磷酸钙磷灰石包被的培养板上培养的TRAP阳性细胞对H（2）S诱导的磷酸钙的吸收显著增加（P <0.01）。H（2）S可促进ERK 1/2和p38的磷酸化，并随时间延长而增加。ERK 1/2和p38的特异性抑制剂PD 98059和SB 203580抑制H（2）S对这些酶的激活和破骨细胞的分化。结论口腔空气中生理浓度的硫化氢可诱导RAW 264细胞产生破骨细胞。
BACKGROUND Hydrogen sulfide (H(2)S), the main component of halitosis, is one of the etiologic factors for periodontitis. We recently reported that H(2)S may induce pathologic changes in rat alveolar bone. The objective of this study is to determine the effect of H(2)S on osteoclast differentiation. METHODS Murine macrophage cells RAW264 were cultured in medium lacking nuclear factor κB ligand (receptor activator of nuclear factor κB ligand) in 5% CO(2) with air at 37°C for 24 hours; then 0.05, 0.5, or 5 ng/ml H(2)S was added to the CO(2)-air mix for 4 days. The controls received the CO(2)-air mix with no H(2)S. Cell differentiation was evaluated by counting the tartrate-resistant acid-phosphatase (TRAP)-positive cells. Extracellular signaling-regulated kinase1/2 (ERK1/2) and mitogen-activated protein kinase p38 phosphorylation were examined by Western blotting. The bone-resorption activity was determined with the resorption assay of calcium phosphate. RESULTS There were significantly more TRAP-positive cells at a concentration of 0.05 ng/ml H(2)S than at the other concentrations (P <0.001). Cathepsin K protein, a specific marker for osteoclasts, was expressed in the H(2)S-induced multinuclear cells. Resorption of calcium phosphate significantly increased in the H(2)S-induced TRAP-positive cells cultured on plates coated with calcium phosphate apatite (P <0.01). The phosphorylation of ERK1/2 and p38 were accelerated by H(2)S, and increased with time. PD98059 and SB203580, specific inhibitors of ERK1/2 and p38, suppressed the activation of these enzymes and osteoclast differentiation by H(2)S. CONCLUSION Results demonstrate that H(2)S at physiologic concentrations in mouth air induces osteoclasts from RAW264 cells.