Dioscorea opposita Thunb has the effect of anti-osteoporosis, but whether its active ingredient diosgenin (DIO) has an anti-osteoporosis effect is unknown. The purpose of this study is to investigate the effect of DIO on the proliferation and differentiation of MG-63 cells. MG-63 cells were treated with different concentrations of DIO (0.001, 0.01, 0.1 and 1 μM) or 20 mM Wnt/β-catenin signaling agonist-LiCl, and then their cell cycle and viability were analyzed by flow cytometry and 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT), respectively. To investigate osteoblast differentiation, alizarin red staining and ultraviolet spectrophotometer were used to determine the number of calcified nodules and the activity of alkaline phosphatase (ALP), respectively. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting were used to detect the expressions of proliferation-related, osteogenic-related and Wnt/β-catenin signal pathway-related factors. After the cells were treated with low-concentration (0.001 or 0.01 μM) DIO, cell viability was significantly increased and the proportion of cells in S phase was increased. In addition, low-concentration DIO could significantly increase the expression of Ki67, proliferating cell nuclear antigen (PCNA), osteopontin (OPN), and osteocalcin (BGP), promote osteoblast differentiation, and suppress the expression of β-catenin, Runx2 and cyclinD1. However, high concentrations of DIO showed the opposite effect. Low-concentration DIO obviously reversed the effect of LiCl on decreasing the number of calcified nodules and inhibiting the expression of OPN and BGP in cells. Low-concentration DIO might promote the proliferation and differentiation of MG-63 cell by inhibiting the Wnt/β-catenin signal pathway.

薯蓣Thunb具有抗骨质疏松作用，但其活性成分薯蓣皂苷元（DIO）是否具有抗骨质疏松作用尚不清楚。本研究的目的是研究DIO对MG-63细胞增殖和分化的影响。MG-63 细胞用不同浓度的 DIO（0.001、0.01、0.1 和 1 μM）或 20 mM Wnt/β-catenin 信号激动剂-LiCl 处理，然后通过流式细胞术和 3-(分别为 4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT)。为了研究成骨细胞分化，使用茜素红染色和紫外分光光度计分别测定钙化结节的数量和碱性磷酸酶（ALP）的活性。采用定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹法检测增殖相关、成骨相关和Wnt/β-catenin信号通路相关因子的表达。细胞经低浓度（0.001或0.01 μM）DIO处理后，细胞活力显着提高，S期细胞比例增加。此外，低浓度DIO可显着增加Ki67、增殖细胞核抗原（PCNA）、骨桥蛋白（OPN）和骨钙素（BGP）的表达，促进成骨细胞分化，抑制β-catenin、Runx2和cyclinD1的表达。 . 然而，高浓度的 DIO 显示出相反的效果。低浓度DIO明显逆转LiCl减少钙化结节数量和抑制细胞内OPN和BGP表达的作用。低浓度DIO可能通过抑制Wnt/β-catenin信号通路促进MG-63细胞的增殖和分化。