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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


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