华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

Notch信号促进核因子κB受体活化因子配体诱导的破骨细胞分化的体外研究

何飞 吴勇 周艳   

  1. 深圳市人民医院口腔科,深圳 518020
  • 出版日期:2015-02-01 发布日期:2015-02-01
  • 通讯作者: 何飞,副主任医师,博士,E-mail:hefeixqkq@aliyun.com
  • 作者简介:何飞,副主任医师,博士,E-mail:hefeixqkq@aliyun.com
  • 基金资助:

    深圳市科技计划基金资助项目(201202108)

Notch signaling promotes receptor activator of nuclear factor kappa B ligand-induced ostoclastogenesis of RAW264.7 cells in vitro

He Fei, Wu Yong, Zhou Yan   

  1. Dept. of Stomatology, Shenzhen People’s Hospital, Shenzhen 518020, China
  • Online:2015-02-01 Published:2015-02-01

摘要:

目的  探讨Notch信号抑制对核因子κB受体活化因子配体(RANKL)诱导的小鼠RAW264.7细胞向破骨细胞分化的影响。方法  建立RANKL诱导的小鼠RAW264.7细胞体外分化模型,实时定量聚合酶链反应(real-time PCR)检测Notch信号成分(Notch1、Notch2、Delta1、Jagged1)、下游靶基因Hes1以及破骨细胞标志基因抗酒石酸酸性磷酸酶(TRAP)和Cathepsin K在诱导前后mRNA的表达。在诱导体系中加入不同浓度的γ分泌酶抑制剂(GSI),抑制Notch受体的表达,TRAP染色检测破骨细胞分化的变化情况。结果  50 ng•mL-1 RANKL诱导小鼠RAW264.7细胞3 d,Notch1、Notch2、Delta1、Jagged1及Hes1的mRNA表达均有不同程度的提高,其中以Notch2、Jagged1增高最明显;破骨细胞标志基因表达显著增高。在RANKL诱导的同时加入不同浓度GSI,抑制Notch的表达,可致Notch下游靶基因Hes1表达下降,同时TRAP阳性细胞计数显著减少,且呈剂量依赖性。结论  Notch信号可促进RANKL诱导的RAW264.7细胞向破骨细胞分化。

关键词: Notch信号, 核因子&kappa, B受体活化因子配体, 破骨细胞分化

Abstract:

Objective  This study aims to explore the effect of Notch signaling depression on the receptor activator of nuclear factor kappa B ligand (RANKL)-induced osteoclastogenesis of RAW264.7 cells. Methods  Mice RAW264.7 cells were cultured and differentiated into osteoclasts with the induction of RANKL. The expressions of Notch1, Notch2, Delta1, Jagged1, Hes1, tartrate-resistant acid phosphatase (TRAP), and Cathepsin K genes during osteoclastogenesis were analyzed using real-time polymerase chain reaction. Osteoclast formation was analyzed using TRAP assay with suppression of Notch receptors by a selective γ-secretase inhibitor (GSI). Results   Notch1, Notch2, Delta1, Jagged1, and Hes1 expressions in RAW264.7 cells were upregulated following 50 ng•mL-1 RANKL stimulation for 3 d, concomitant with the expression of the osteoclast differentiation markers TRAP and Cathepsin K. Notch2 and Jagged1 had the most remarkable increase in the Notch family members. GSI inhibited RANKL-induced osteoclastogenesis of RAW264.7 cells and Hes1 expression dosedependently. Conclusion  Notch signaling activation may promote RANKL-induced osteoclastogenesis of RAW264.7 cells.

Key words:  Notch signaling, receptor activator of nuclear factor kappa B ligand, osteoclastogenesis