华西口腔医学杂志

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压应力对小鼠单核细胞RAW264.7 DNAX活化蛋白12、抗酒石酸酸性磷酸酶表达的影响

黄生高  凌天牖  钟孝欢  刘云峰   

  1. 中南大学湘雅二医院口腔中心,长沙 410011
  • 出版日期:2013-08-01 发布日期:2013-08-01
  • 通讯作者: 黄生高,Tel: 13973197181
  • 作者简介:黄生高(1964—),男,湖南人,副教授,博士

Effect of compressive stress on the expression of DNAX-activating protein 12 and tartrate-resistant acid phosphatase in mouse monocyte RAW264.7

Huang Shenggao, Ling Tianyou, Zhong Xiaohuan, Liu Yunfeng.   

  1. Dept. of Stomatology, The Second Xiangya Hospital of Central South University, Changsha 410011, China
  • Online:2013-08-01 Published:2013-08-01

摘要:

 目的   探讨压应力对小鼠单核细胞RAW264.7 DNAX活化蛋白12(DAP12)、抗酒石酸酸性磷酸酶(TRAP)表达的影响。方法   以小鼠单核细胞RAW264.7为研究对象,采用四点弯曲体外细胞加载装置加载压应力0、3、6、12 h,分别以逆转录聚合酶链反应(RT-PCR)、蛋白免疫印迹法检测DAP12、TRAP mRNA和DAP12蛋白的表达情况。结果   小鼠单核细胞RAW264.7经破骨细胞培养液培养后,体积变大,核数目增多,TRAP染色阳性。受压应力刺激后,DAP12、TRAP mRNA及DAP12蛋白表达随加力时间延长而增加(P<0.05)。结论   小鼠单核细胞RAW264.7经破骨细胞培养液培养后成功向破骨细胞转化,转化细胞受压应力刺激活化过程中存在DAP12高表达。

关键词:  , DNAX活化蛋白12, 抗酒石酸酸性磷酸酶, 压应力, 破骨细胞

Abstract:

 Objective   To investigate the expression of DNAX-activating protein 12(DAP12) and tartrate-resistantacid phosphatase(TRAP) in mouse monocyte RAW264.7 subjected to compressive stress. Methods   Mouse monocyteRAW264.7 was subjected to four-point bending system. The expression of DAP12 and TRAP mRNA were examined byreverse transcription-polymerase chain reaction(RT-PCR) and the expression of DAP12 protein by Western Blottingafter 0, 3, 6 and 12 h’s compressive stress. Results   After RAW264.7 cells were cultured by osteoclast cell culturefluid, the amount of the cells increased, and the volume enlarged, and the number of nuclei per osteoclast increasedin vitro. The expression of DAP12 and TRAP mRNA and DAP12 protein of RAW264.7 cells subjected to compressivestress increased along with the time(P<0.05). Conclusion  The mouse monocyte RAW264.7 can differentiate intoosteoclast in vitro, and high expression of DAP12 exists in the process of osteoclast differentiation.

Key words:  DNAX-activating protein 12, tartrate-resistant acid phosphatase, compressive stress, osteoclast