›› 2012, Vol. 32 ›› Issue (10): 1302-.doi: 10.3969/j.issn.1674-8115.2012.10.005

• Original article (Basic research) • Previous Articles     Next Articles

Molecular mechanisms of substance P-induced monocyte chemoattractant protein-1 secretion in fibroblasts under high glucose culture condition

JIA Zhi-gang1,2, FANG Yong1,2, YAO Min1,2, YU Wei-rong1, XU Peng1, JIA Yi-tao2, NI Tao1,2   

  1. 1.Department of Burns and Plastic Surgery, the Third People´s Hospital, 2.Institute of Traumatic Medicine, Shanghai Jiaotong University School of Medicine, Shanghai 201900, China
  • Online:2012-10-28 Published:2012-11-05
  • Supported by:

    Shanghai Municipal Health Bureau Foundation, 2009102；Foundation of the Third People´s Hospital, Shanghai Jiaotong University School of Medicine, syz2011-13

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Abstract:

Objective To investigate the molecular mechanisms of substance P (SP)-induced monocyte chemoattractant protein-1 (MCP-1) secretion in fibroblasts under high glucose culture condition. Methods Human skin fibroblasts were cultured in high glucose DMEM medium, and were incubated with 10 nmol/L SP for different time. The expression of inhibitor α of nuclear factor kappa B (NF-κB)(IκBα), phosphonated IκBα (p-IκBα) and phosphonated NF-κB subunit p65 (p-NF-κBp65) in cells was detected by Western blotting, and the expression and translocation of p-NF-κBp65 was observed by immunofluorescence method. Human skin fibroblasts cultured in high glucose DMEM medium were divided into control group, SP group and SP+NF-κB inhibitor (MG132) group (pretreatment with MG132 for 60 min), the expression of p-NF-κBp65 in cells was detected in each group, and the concentration of MCP-1 in the supernatant was determined by ELISA. Results The expression of IκBα significantly decreased, and that of p-IκBα significantly increased in fibroblasts with the time increase of treatment with SP (P<0.05). The expression of p-NF-κBp65 significantly increased after treatment with SP (P<0.05), which was in a time-dependent manner. The expression of p-NF-κBp65 in nuclei significantly increased after treatment with SP observed under laser confocal microscope with immunofluorescence staining. The expression of p-NF-κBp65 in cells and the concentration of MCP-1 in the supernatant in SP+MG132 group were significantly lower than those in control group (P<0.05). Conclusion The mechanisms of SP-induced MCP-1 secretion in fibroblasts may be related to the activation of NF-κB signalling pathway.

Key words: substance P, human skin fibroblasts, nuclear factor kappa B, monocyte chemoattractant protein-1