Role of Mechanical Stress-induced Glutamate Signaling-associated Molecules in Cytodifferentiation of Periodontal Ligament Cells
Role of Mechanical Stress-induced Glutamate Signaling-associated Molecules in Cytodifferentiation of Periodontal Ligament Cells
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DOI：
10.1074/jbc.m109.097303
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发表时间：
2010-09-03
影响因子：
4.8
通讯作者：
Murakami, Shinya
中科院分区：
文献类型：
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作者：
Fujihara, Chiharu;Yamada, Satoru;Murakami, Shinya
在这项研究中，我们分析了拉伸机械应力对体外培养的人牙周膜（PDL）细胞基因表达谱的影响。DNA芯片分析鉴定了17个在机械应力下人PDL细胞中上调的基因，包括HOMER 1（homer homolog 1）和GRIN 3A（glutamate receptor ionotropic N-methyl-D-aspartate 3A），它们与谷氨酸信号传导有关。RT-PCR和实时PCR分析表明，人PDL细胞组成型表达谷氨酸信号相关基因，机械应力增加这些mRNA的表达，导致谷氨酸从人PDL细胞和细胞内谷氨酸信号转导的释放。有趣的是，外源性谷氨酸在PDL细胞分化过程中增加了细胞分化和矿化相关基因的RNA以及ALP（碱性磷酸酶）活性。另一方面，谷氨酸信号传导抑制剂利鲁唑和（+）-MK 801马来酸盐抑制碱性磷酸酶活性和矿化结节的形成过程中的细胞分化和矿化。阿曲唑抑制机械应力诱导的人PDL细胞谷氨酸信号相关基因的表达。此外，原位杂交分析显示，上调谷氨酸信号相关的基因表达的张力网站在牙周膜正畸牙齿移动的小鼠模型。目前的数据表明，由机械应力诱导的谷氨酸信号正向调节PDL细胞的细胞分化和矿化。
In this study, we analyzed the effects of tensile mechanical stress on the gene expression profile of in vitro-maintained human periodontal ligament (PDL) cells. A DNA chip analysis identified 17 up-regulated genes in human PDL cells under the mechanical stress, including HOMER1 (homer homolog 1) and GRIN3A (glutamate receptor ionotropic N-methyl-D-aspartate 3A), which are related to glutamate signaling. RT-PCR and real-time PCR analyses revealed that human PDL cells constitutively expressed glutamate signaling-associated genes and that mechanical stress increased the expression of these mRNAs, leading to release of glutamate from human PDL cells and intracellular glutamate signal transduction. Interestingly, exogenous glutamate increased them RNAs of cytodifferentiation and mineralization-related genes as well as the ALP (alkaline phosphatase) activities during the cytodifferentiation of the PDL cells. On the other hand, the glutamate signaling inhibitors riluzole and (+)-MK801 maleate suppressed the alkaline phosphatase activities and mineralized nodule formation during the cytodifferentiation and mineralization. Riluzole inhibited the mechanical stress-induced glutamate signaling-associated gene expressions in human PDL cells. Moreover, in situ hybridization analyses showed up-regulation of glutamate signaling-associated gene expressions at tension sites in the PDL under orthodontic tooth movement in a mouse model. The present data demonstrate that the glutamate signaling induced by mechanical stress positively regulates the cytodifferentiation and mineralization of PDL cells.