上海交通大学学报(医学版)

• 论著(基础研究) • 上一篇    下一篇

神经肽P物质通过诱导RUNX2表达对小鼠成骨细胞增殖能力的影响

余炜,朱超,茅伟伟,赵恩典,孟胜伟,蒋盛旦   

  1. 上海交通大学 医学院附属新华医院骨科,上海 200092
  • 出版日期:2017-01-28 发布日期:2017-01-19
  • 通讯作者: 蒋盛旦,电子信箱:spine_jiangsd@126.com。
  • 作者简介:余炜(1990—),男,硕士生;电子信箱:halouyuwei@163.com。
  • 基金资助:

    上海市优秀青年医学人才培养计划(XYQ2011026)

Effect of neuropeptide substance P on proliferation of murine osteoblasts through induction of RUNX2 expression

YU Wei, ZHU Chao, MAO Wei-wei, ZHAO En-dian, MENG Sheng-wei, JIANG Sheng-dan   

  1. Department of Orthopedics Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China
  • Online:2017-01-28 Published:2017-01-19
  • Supported by:

    Excellent Young Medical Talents Training Program of Shanghai Municipality,XYQ2011026

摘要:

目的 ·探讨神经肽P物质(SP)对转录因子RUNX2表达的调控及其对成骨细胞增殖能力的影响。方法 ·体外分离C57BL/6J小鼠颅骨来源的原代成骨细胞,通过转染siRNA沉默成骨细胞RUNX2基因的表达。将SP、Spantide(SP抑制剂)、L703606(NK-1受体抑制剂)以及RUNX2 siRNA进行配伍作用于成骨细胞,观察SP对成骨细胞RUNX2表达的调控及对成骨细胞增殖能力的影响。成骨细胞增殖活性的检测采用细胞增殖-毒性检测试剂盒(CCK-8法)。通过real-time PCR和Western blotting技术检测SP对RUNX2在mRNA和蛋白表达水平的调控。结果 · CCK-8法检测结果显示,浓度为10-12~10-6 mol/L的SP促进小鼠成骨细胞增殖,10-8 mol/L为其最佳浓度。10-8 mol/L SP作用于成骨细胞48 h后,转录因子RUNX2的mRNA和蛋白表达水平较对照组明显升高,Spantide和L703606抑制SP对RUNX2表达的诱导。RUNX2 siRNA降低成骨细胞基础增殖水平,同时也抑制了SP对成骨细胞的促增殖作用。结论 · SP通过与NK-1受体结合诱导转录因子RUNX2的表达,进而增强成骨细胞增殖能力。

关键词: 神经肽P物质, 成骨细胞, 转录因子RUNX2, NK-1受体

Abstract:

Objective · To investigate the effects of neuropeptide substance P(SP) on RUNX2 expression as well as cell proliferation in osteoblasts. Methods · Primary cultured murine skull-derived osteoblasts were isolated from C57BL/6J mice;gene expression of RUNX2 in osteoblast was silenced by transfection with siRNA. Osteoblasts were treated with SP,with or without Spantide(specific blocking agents of SP),L703606(specific blocking agents of NK-1 receptor)and RUNX2 siRNA,then the effects of SP on RUNX2 expression and cell proliferation were investigated. Osteoblast proliferation was determined by cell proliferation-toxicity assay kit(CCK-8) assay. RUNX2 expressions at the mRNA and protein levels were analyzed by real-time PCR and Western blotting analysis,respectively. Results · CCK8 assay showed that SP at the concentration of 10-12~10-6 mol/L promoted osteoblasts proliferation, with 10-8 mol/L being the most effective concentration. The expressions of RUNX2 at the mRNA and protein levels were significantly increased after osteoblasts were treated with 10-8 mol/L SP for 48 h. Moreover,Spantide and L703606 inhibited SP-induced enhancement of RUNX2 expression;RUNX2 siRNA reduced the baseline of osteoblast proliferation and further attenuated the promoting effects of SP on osteoblast proliferation. Conclusion · SP induced RUNX2 expression through binding to NK-1 receptor,thus promoting the proliferation of osteoblasts.

Key words: substance P, osteoblast, RUNX2, NK-1 receptor