Original article (Basic research)

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
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  • Department of Orthopedics Surgery, Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China

Online published: 2017-01-19

Supported by

Excellent Young Medical Talents Training Program of Shanghai Municipality,XYQ2011026

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.

Cite this article

YU Wei , ZHU Chao , MAO Wei-wei , ZHAO En-dian , MENG Sheng-wei , JIANG Sheng-dan . Effect of neuropeptide substance P on proliferation of murine osteoblasts through induction of RUNX2 expression[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2017 , 37(1) : 15 . DOI: 10.3969/j.issn.1674-8115.2017.01.004

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