重楼皂苷促大鼠成骨细胞增殖的作用及其机制研究

Effect of Polyphyllins on Promoting Proliferation of Rats Osteoblasts and the Possible Mechanisms

目的观察重楼皂苷对骨质疏松大鼠及其成骨细胞的作用并探索其可能作用机制。方法采用摘除卵巢(ovariectomy,OVX)的方法建立大鼠骨质疏松模型,观察重楼皂苷对该模型的骨质疏松保护作用;使用多次酶消化法分离培养大鼠原代成骨细胞,加入不同浓度的重楼皂苷(3、10、30、100μg/ml),采用CCK-8试剂盒、碱性磷酸酶(alkaline phosphatase,ALP)活性检测法观察成骨细胞的增殖及分化情况,Western blot方法观察不同浓度重楼皂苷对大鼠成骨细胞中骨保护蛋白(osteoprotegerin,OPG)/核因子-κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)表达的影响。结果重楼皂苷可以有效地改善骨质疏松大鼠股骨干骺端骨小梁的微结构特性;可有效促进大鼠原代成骨细胞的增殖,浓度依赖性地上调OPG的表达,下调RANKL的表达。结论重楼皂苷可通过调节OPG/RANKL信号通路促进大鼠原代成骨细胞增殖与分化。

Objective To evaluate the ability of polyphyllins to prevent osteoporosis induced by ovariectomy(OVX)in rats,and assessed its effect on the proliferation of primary osteoblasts of rats and the possible mechanisms.Methods The effect of polyphyllins on the OVX-induced osteoporosis in rats was observed.Primary cultures of rats' osteoblasts were obtained by enzyme digestion and administered with different concentrations of polyphyllins(3,10,30,and 100μg/ml).Then,CCK8 cell counting kit,and alkaline phosphatase(ALP)activity detection were used to observe the proliferation of primary osteoblasts of rats.Western blot analysis was used to evaluate the expression of osteoprotegerin(OPG)and receptor activator of nuclear factor-κB ligand(RANKL)in primary osteoblasts.Results Polyphyllins could effectively alleviated trabecular microarchitectural properties,promoted proliferation of primary osteoblasts,upregulated the expression of OPG,and downregulated RANKL expression.Conclusions Polyphyllins may enhance primary osteoblasts of rat proliferation and differentiation through regulating OPG/RANKL signal pathway.