葛根素抑制1，25-二羟基维生素D3促破骨细胞骨吸收功能的最佳剂量

背景：近年来国内外学者对葛根素的骨形成研究较多,但对于葛根素抑制1,25-(OH)₂D₃促破骨细胞骨吸收功能尚未见报道。 目的：观察不同浓度葛根素在体外对1,25-(OH)₂D₃促破骨细胞骨吸收功能的抑制作用。 方法：收集3周龄小鼠骨髓细胞并接种到24孔培养板中。实验组培养液为1,25-(OH)₂D₃ +α-MEM完全培养基+不同质量浓度的葛根素,对照组不加葛根素。 结果与结论：不同浓度组葛根素处理的培养体系均诱导出破骨样细胞且形态差异不明显;除100 μg/L葛根素组在第3天时碱性磷酸酶表达显著增高外,各组之间培养液上清中碱性磷酸酶水平无显著性差异(P > 0.05);10 μg/L葛根素组培养液上清Ca2+水平在第3,5,12天时较对照组显著降低(P < 0.05~0.01),50 μg/L葛根素组培养液上清Ca²⁺水平在第12天时较对照组显著降低(P < 0.01),100 μg/L葛根素组培养液上清Ca²⁺水平在所有检测点较10 μg/L葛根素组显著上升(P < 0.05)。提示葛根素最佳抑制剂量在10~50 μg/L之间。

Inhibitory effect of puerarin on osteoclastic bone resorption induced by 1,25-dihydroxyvitamin D3 in vitro: The optimal concentration of puerarin

BACKGROUND:Many studies have focused on the effect of puerarin on the bone formation. However, the inhibitory effect of puerarin on osteoclastic bone resorption induced by 1,25-dihydroxyvitamin D₃ in vitro remain poorly understood. OBJECTIVE:To study the inhibitory effect of different concentrations of puerarin on osteoclastic bone resorption induced by 1,25-dihydroxyvitamin D3 in vitro. METHODS:Bone marrow cells were isolated from 3-week-old mice and added to 24-well plates. The cells in the experiment groups were cultured in 1,25-dihydroxyvitamin D₃+10%+alpha-modified minimum Eagles medium+various concentrations of puerarin (10, 50 and 100 μg/L). The control group was not treated with puerarin. RESULTS AND CONCLUSION:Osteoclast-like cells were formed and were not significantly different among the four groups. As to the concentration of alkaline phosphatase in the supernatant liquid, there were no significant differences among the four groups (P > 0.05), but the concentration of alkaline phosphatase in the 100 μg/L puerarin group was significantly higher than that of 0 and 50 μg/L puerarin on day 3 (both P < 0.01). The concentration of Ca²⁺ in the 10 μg/L puerarin group was significantly lower than that in the control group on days 3, 5 and 12 (P < 0.05-0.01), the same as the 50 μg/L puerarin group on day 12 (P < 0.01). Compared with the 10 μg/L puerarin group, the concentration of Ca²⁺ in the 100 μg/L puerarin group was significantly higher for up to day 12 (P < 0.05). The results demonstrat that the best dose of puerarin which has the inhibitory effect on osteoclastic bone resorption induced by 1,25-dihydroxyvitamin D₃ in vitro is between 10-50 μg/L.