目的探讨骨灵片促进成骨细胞增殖、矿化以及基因表达的细胞信号通路。方法用含有骨灵片的大鼠血清、p38MAPK抑制剂SB203580作用人成骨细胞MG-63,测定各组成骨细胞增殖率、矿化结节数量。采用Western-blotting及RealtimeRT-PCR方法观察各组成骨细胞护骨素(OPG)、护骨素配体(RANKL)以及巨噬细胞集落刺激因子(M-CSF)基因表达和磷酸化p38含量。结果经含骨灵片药液的大鼠血清作用后,MG-63细胞的增殖和矿化结节形成数明显增加,OPG基因表达增加,RANKL基因表达减少,OPG/RANKL比值显著增加,M-CSF有下降趋势,但没有显著性差异,同时刺激p38磷酸化。结论骨灵片可能通过p38MAPK通路促进成骨细胞增殖以及矿化结节形成,调控OPG、RANKL以及M—CSF的表达,达到治疗骨质疏松症的作用。
Objective To investigate the effect of Gulingpian (GLP, or Guling tablets) on the proliferation and mineralization of MG-63 cells and the involvement of p38 MAPK pathway in such processes. Methods MG-63 cells were cultured in the presence of p38 inhibitor SB203580 and/or serum from rats fed with normal saline or GLP. The expression of the OPG, RANKL and M-CSF mRNA, cell proliferation and formation of mineralized bone nodule, the total protein of p38 and phosphorylated p38, were observed or quantitatively measured. Results Treatment with GLP serum increased the level of p38 phosphorylation but did not affect the total p38 expression. GLP serum significantly promoted MG-63 cell proliferation and differentiation, resulting also in up-regulated OPG and down-regnlated RANKL mRNA expressions without obvious alteration in M-CSF expression. These effects were blocked by the specific p38 inhibitor SB203580. Conclusion GLP promotes the proliferation and differentiation ofosteoblasts and regulates OPG/RANKL expressions in vitro, which are mediated probably via the p38 MAPK pathway.