摘要/Abstract

摘要： 目的 ·探索细胞核因子 κB受体激活蛋白配体（receptor activator of nuclear factor-κB ligand，RANKL）/核因子 κB受体激活蛋白（receptor activator of nuclear factor-κB，RANK）系统及 Notch通路间的交互调控机制。方法 ·通过 RANKL（35 ng/L）刺激 RAW264.7小鼠破骨细胞前体细胞系，用 real-time PCR、Western blotting检测 Notch通路关键分子的表达水平。小干扰 RNA（small interfering RNA，siRNA）技术抑制 Notch通路中 Notch1、Dll1和 Dll3分子的表达，检测上述分子抑制后 Rank mRNA和蛋白表达的变化。结果 · RANKL刺激引起 Notch1、Jagged1、Jagged2的表达降低， Dll4的表达升高。 RANKL抑制 Notch通路下游分子 Hes1、Hey1的表达。通过 siRNA技术，Notch1、Dll1和 Dll3 mRNA的表达分别下调 71%（P0.000）、53%（P0.015）和 70%（P0.000）。Notch1表达抑制后，Rank mRNA和蛋白的表达均增加（P0.003，P0.000）。结论 · Notch1对 RANKL-RANK系统的潜在调节机制，可能在破骨细胞相关的溶骨活动中发挥重要作用。
关键词: 破骨细胞, 溶骨, Notch通路, RANKL-RANK系统
Abstract:
Objective · To investigate the crosstalk between receptor activator of nuclear factor-κB ligand (RANKL) / receptor activator of nuclear factorκB (RANK) system and Notch signaling pathways. Methods · Moosteoclast precursor cell line RAW264.7 was cultured and stimulatedRANKL (35 ng/L). Real-time PCR and Western blotting analysis were used to determine the s of Notch signaling molecules in RANKL-stimulated RAW264.7 cells. Small interfering RNA (siRNA) transfection was used to inhibit the of Notch1, Dll1 and Dll3 in Notch signaling pathways, then the mRNA and protein s of Rank were detected using real-time PCR and Western blotting analysis, respectively. Results · The s of Notch1, Jagged1 and Jagged2 were down-regulated while the of Dll4 was up-regulated after RANKL stimulation. The stimulation also inhibited the of Hes1 and Hey1. After siRNA transfection, the mRNA s of Notch1, Dll1 and Dll3 were suppressed71% (P0.000), 53% (P0.015) and 70% (P0.000), respectively. The mRNA and protein s of Rank were up-regulated after Notch1 inhibition (P0.003, P0.000). Conclusion · The data infers that the impact of Notch1 on RANKL/RANK system might play a key role in bone resorption conductedosteoclast.
Key words: osteoclast, bone resorption, Notch pathway, RANKL/RANK system


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