Effect of the release from mechanical stress on osteoclastogenesis in RAW264.7 cells
Effect of the release from mechanical stress on osteoclastogenesis in RAW264.7 cells
复制标题
DOI：
10.3892/ijmm.2011.675
复制
发表时间：
2011-07-01
影响因子：
5.4
通讯作者：
Iida, Junichiro
中科院分区：
文献类型：
--
作者：
Shibata, Kenjiro;Yoshimura, Yoshitaka;Iida, Junichiro
机械应力释放对破骨细胞形成的影响可能与施加机械应力一样重要。然而，机械应力对破骨细胞行为的直接影响还没有得到彻底的研究，关于机械应力释放的结果的信息也有限。在这项研究中，检测了机械应力的施加和释放对破骨细胞分化的影响。检测RAW264.7细胞来源的抗酒石酸酸性磷酸酶(TRAP)阳性多核破骨细胞的数量，并用实时荧光定量聚合酶链式反应(Real-time PCR)检测破骨细胞分化基因的表达，这些基因在机械应力释放后根据Flexforce细胞张力系统发生改变。机械应力作用可抑制破骨细胞的分化和融合，机械应力释放后可迅速诱导破骨细胞分化和融合。破骨细胞特异性基因TRAP、基质金属蛋白酶-9(MMP9)、组织蛋白酶K(cath-k)、降钙素受体(CTR)、ATPase H+转运空泡质子泵成员I(ATP6i)、氯通道-7(ClC7)和树突状细胞特异性跨膜蛋白(DC-STAMP)的表达随机械应力的增加而降低，释放后48h表达增强。这些基因mRNA表达的改变与破骨细胞和大破骨细胞的数量有关。诱导型一氧化氮合酶(INOS)mRNA随机械应力的增加而增加，释放后下降。一氧化氮(NO)的产生随机械应力的增加而增加。活化T细胞胞浆核因子(NFATc)家族的mRNAs在机械应力作用下无明显变化，但在释放后48h表达上调。这些结果表明，机械应力抑制破骨细胞分化和融合是通过诱导型一氧化氮合酶(INOS)升高的结果，机械应力释放后促进破骨细胞分化和融合与NFATc家族基因有关，NFATc家族基因在机械应力作用下表达不变，但在机械应力释放后表达上调。
The effects of mechanical stress release on osteoclastogenesis may be as Important as those of mechanical stress application. However, the direct effects of mechanical stress on the behavior of osteoclasts has not been thoroughly investigated and there is limited information on the results of the release from mechanical stress. In this study, the effects of mechanical stress application and its release on osteoclast differentiation were examined. The number of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated osteoclasts derived from RAW264.7 cells were measured and the expression of osteoclast differentiation genes, which was altered in response to the release from mechanical stress according to the Flexercell tension system was evaluated by real-time PCR. Osteoclast differentiation and fusion were suppressed by mechanical stress application and were rapidly induced after mechanical stress release. The mRNA expression of the osteoclast specific genes, TRAP, matrix metalloproteinase-9 (MMP-9), cathepsin-K (cath-k), calcitonin receptor (CTR), ATPase H+ transporting vacuolar proton pump member I (ATP6i), chloride channel-7 (ClC7) and dendritic cell-specific transmembrane protein (DC-STAMP) was decreased with mechanical stress application, and increased up to 48 h after the release from it. These alterations in gene mRNA expression were associated with the number of osteoclasts and large osteoclasts. Inducible nitric oxide synthetase (iNOS) mRNA was increased with mechanical stress and decreased after its release. Nitric oxide (NO) production was increased with mechanical stress. Nuclear factor of activated T cells cytoplasmic (NFATc) family mRNAs were not altered with mechanical stress, but were up-regulated up to 48 h after the release from it. These findings indicate that the suppression of osteoclast differentiation and fusion induced by mechanical stress is the result of NO increase via iNOS, and that the promotion of osteoclast differentiation and fusion after the release from mechanical stress is related to the NFATc family genes, whose expression remained constant during mechanical stress but was up-regulated after the release from mechanical stress.