川续断皂苷Ⅵ对小鼠成肌细胞成骨分化的影响

目的 探讨川续断皂苷Ⅵ对小鼠成肌细胞成骨分化的作用及影响。方法 配制浓度为1×10-8～1×10-3 mol/L的川续断皂苷Ⅵ药物溶液，用细胞计数试剂盒8检测其对小鼠成肌细胞的细胞毒性和增殖能力。然后用碱性磷酸酶活性分析确定促进其成骨分化的最佳浓度。最后采用免疫印迹分析、实时定量PCR和茜素红染色观察成骨分化效果。结果 实验结果表明，1×10-3、1×10-4 mol/L川续断皂苷Ⅵ对小鼠成肌细胞有明显的毒性作用（P＜0.01）。1×10-8～1×10-5 mol/L对细胞增殖无明显作用。1×10-6 mol/L川续断皂苷Ⅵ促进小鼠成肌细胞成骨分化作用最为明显（P＜0.01）。与对照组相比，1×10-6 mol/L川续断皂苷Ⅵ组的ALP、Runx2、Ocn和Col1α1的mRNA及蛋白表达量均明显升高（P＜0.05），茜素红染色也显示有更多的钙结节形成。结论 1×10-6 mol/L川续断皂苷Ⅵ能明显促进小鼠成肌细胞的成骨分化，但对其增殖无明显影响。

Effects of Asperosaponin Ⅵ on osteogenic differentiation of mouse myoblasts

Objective To investigate the effects of Asperosaponin Ⅵ on osteogenic differentiation of mouse myoblasts. Methods The concentrations of Asperosaponin Ⅵ were 1×10-3–1×10-8 mol/L. Cell counting kit 8 was used to detect the cytotoxicity and proliferation ability of Asperosaponin Ⅵ on mouse myoblasts. The optimal concentration of ALP was determined by alkaline phosphatase activity analysis. Finally, Western blot analysis, real-time quantitative PCR and alizarin red staining were used to observe the effect of osteogenic differentiation. Results The results showed that 1×10-3、1×10-4 mol/L Asperosaponin Ⅵ had obvious toxic effects on mouse myoblasts (P<0.01), while 1×10-5–1×10-8mol/L had no obvious effect on cell proliferation. It was also shown that 1×10-6mol/L Asperosaponin Ⅵ significantly promoted the osteogenic differentiation of mouse myoblasts (P<0.01). Compared with the control group, the mRNA and protein expressions of ALP, Runx2, Ocn and col1 α 1 in the concentration of 1×10-6 mol/L Asperosaponin Ⅵ were significantly increased (P<0.05), and alizarin red staining also showed more calcium nodules. Conclusion 1×10-6mol/L Asperosaponin Ⅵ can significantly promote the osteogenic differentiation of mouse myoblasts, but has no obvious effect on its proliferation.