摘要
目的探讨红景天苷对人成骨肉瘤细胞株MG63诱导成骨分化作用及相关分子机制。方法 MTT法测定不同浓度红景天苷对MG63细胞增殖活力的影响;PNPP法检测经红景天苷处理后成骨性标志物——碱性磷酸酶活性的变化;RT-PCR检测细胞内bmp2 mRNA水平的变化;Western blot及流式细胞术检测给药前后细胞内BMP2、IκBα蛋白表达水平的变化。结果红景天苷能显著促进MG63细胞的增殖并上调其ALP活性,其对ALP活性的上调呈现一定的时间和剂量依赖性,其中在5μmol/L、48 h条件下表现出最佳效果,证明红景天苷具有促成骨分化作用。此外,红景天苷能显著上调细胞内bmp2 mRNA及蛋白表达水平,并能时间依赖性激活IκBα,抑制NF-κB活性。结论红景天苷能促进人成骨肉瘤细胞成骨分化,具有潜在的骨代谢调节作用,其作用机制可能与抑制NF-κB活性并激活BMP2信号通路有关。
Objective To investigate the effect of salidroside (SAL) on osteoblast differentiation and related signaling factors in human osteosarcoma cells MG63. Methods Cell viability was assessed by MTT method, and the ALP activity was measured by PNPP method. Gene expression of bmp2 was detected by RT-PCR analysis, and the protein levels of BMP2 and IκBα were determined by Western blot and flow cytometry. Results SAL promoted the proliferation of MG63 cells. In addition, the ALP activity was significantly increased in a time- and dose-dependent manner after SAL treatment, suggesting that SAL could stimulate osteoblast differentiation. Both the bmp2 mRNA and protein expressions were increased after SAL treatment. Meanwhile, SAL could time-dependently increase IκBα expression, which may affect NF-κB activity. Conclusion SAL can stimulate osteoblast differentiation and regulate both NF-κB and BMP2 signaling pathway in MG63 cells, suggesting that the natural product may be developed as a promising drug with little toxicity to regulate bone metabolism.
出处
《中国医药生物技术》
2013年第6期414-419,共6页
Chinese Medicinal Biotechnology
基金
国家自然科学基金(81072645
81328024)
"重大新药创制"国家科技重大专项(2012ZX09301002-001&-003)
