葛根素调控骨代谢的体外实验研究

Effect of puerarin on the bone metabolism in vitro

目的 :探讨中药葛根对骨形成和骨吸收的调控作用。方法 :鼠颅盖骨机械法体外分离培养获得成骨细胞(osteoblast ,OB) ;新生兔长干骨机械分离法获得破骨细胞 (osteoclast ,OC)。实验组培养液为 10 % (体积分数 )胎牛血清 +αMEM培养液 +不同 (质量 )浓度的葛根素 ,对照组不加葛根素。分别用四甲基偶氮唑盐 (MTT)和碱性磷酸酶 (ALP)法检测OB的增殖和分化状态 ;抗酒石酸酸性磷酸酶 (TRAP)染色观察OC形态变化 ,图像分析体外骨吸收陷窝面积 ,原子分光光度计法测定OC培养液上清液中Ca2 + 含量。结果 :中药葛根素对大鼠OB的增殖无明显的刺激作用 ,但能够促进OB合成分泌ALP。同时 ,造成体外培养的OC空泡性变 ,骨吸收陷窝面积减少和培养液上清液中Ca2 + 含量的下降。结论 :葛根通过抑制骨吸收 ,刺激骨形成来调控骨代谢 。

Objective:To study the effect of puerarin on the bone formation and the bone resorption in vitro . Methods:The osteoblasts (OBs) were obtained from the rat calvaria . The osteoclasts (OCs) were isolated by the methods according Chambers' and Yu Shifeng's. The cells in the experiment group were grown in 10% fetal calf serum ( FCS ) + αMEM + various concentrations of puerarin. The control groups were grown in 10% FCS +αMEM. The growth of OBs was assessed by MTT method. The differentiation of OBs was examined by the ALP activity. The OCs were observed by the TRAP stain. The area of bone resorption and the concentration of Ca 2+ in the supernatant liquid of OCs cultures were also tested. Results:Puerarin had no effect on the growth of the OBs, but it could stimulate the formation and the secretion of alkaline phosphatase (ALP). The OCs vacuolarized under the effect of puerarin. The area of bone resorption and the concentration of Ca 2+ in the supernatant liquid of OCs cultures were both decreased. Conclusion:Puerarin can suppress the bone resorption and promote the bone formation, but the former effect is stronger than the latter.