Calycosin stimulates the osteogenic differentiation of rat calvarial osteoblasts by activating the IGF1R/PI3K/Akt signaling pathway
Calycosin stimulates the osteogenic differentiation of rat calvarial osteoblasts by activating the IGF1R/PI3K/Akt signaling pathway
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毛蕊异黄酮通过激活IGF1R/PI3K/Akt信号通路刺激大鼠颅骨成骨细胞成骨分化
DOI：
10.1002/cbin.11102
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发表时间：
2019-03-01
影响因子：
3.9
通讯作者：
Chen, Keming
中科院分区：
文献类型：
--
作者：
Fang, Yaoyao;Xue, Zhiyuan;Chen, Keming
据报道，毛蕊异黄酮具有很强的成骨活性，并与抗骨质疏松作用呈正相关。然而，其确切的作用机制仍不清楚。由于胰岛素样生长因子 1 受体 (IGF1R) 信号传导和磷脂酰肌醇 3-激酶/Akt (PI3K/Akt) 信号传导已被证明在调节成骨中发挥关键作用，因此我们假设毛蕊异黄酮的成骨活性是由这些信号传导途径介导的。大鼠颅骨成骨细胞 (ROB) 在含有毛蕊异黄酮的成骨培养基中培养，含或不含 GSK1904529A (GSK) 或 LY294002 (LY)（分别为 IGF1R 和 PI3K 抑制剂）。对细胞增殖、碱性磷酸酶 (ALP) 活性、钙化结节、成骨基因 [碱性磷酸酶 (Alpl)、I 型胶原蛋白 (Col1a1)、runt 相关转录因子 2 (Runx2)、Osterix 和骨形态发生蛋白 2 (Bmp2)] 的 mRNA 或蛋白表达以及 IGF1R 和骨形态发生蛋白 2 (Bmp2) 的磷酸化的影响 Akt 进行了检查。本研究结果表明，毛蕊异黄酮在10(-8)-10(-6) M范围内以剂量依赖性方式增强细胞增殖、ALP活性和茜素红-S染色，而在10(-5) M时观察到抑制作用。最佳浓度(10(-6) M，生理上可达到的浓度)处理增加了成骨基因的mRNA水平和成骨基因的磷酸化。 IGF1R 和 Akt。此外，GSK 或 LY 治疗部分逆转了毛蕊异黄酮对 ROB 的影响，表现为毛蕊异黄酮诱导的 ALP 活性、钙化结节和成骨基因表达的降低。这些结果表明毛蕊异黄酮的成骨作用部分涉及IGF1R/PI3K/Akt信号通路。
Calycosin has been reported to have a strong osteogenic activity and a positive correlation with anti-osteoporosis effects. However, its precise mechanism of action remains unclear. Since insulin-like growth factor 1 receptor (IGF1R) signaling and phosphatidylinositol 3-kinase/Akt (PI3K/Akt) signaling have been shown to play a pivotal role in regulating osteogenesis, we hypothesized that the osteogenic activity of calycosin is mediated by these signaling pathways. Rat calvarial osteoblasts (ROBs) were cultured in osteogenic medium containing calycosin with or without GSK1904529A (GSK) or LY294002 (LY) (inhibitors of IGF1R and PI3K, respectively). The effects on cell proliferation, alkaline phosphatase (ALP) activity, calcified nodules, mRNA or protein expression of osteogenic genes [alkaline phosphatase (Alpl), collagen type I (Col1a1), runt-related transcription factor 2 (Runx2), Osterix, and bone morphogenetic protein 2 (Bmp2)], and phosphorylation of IGF1R and Akt were examined. The present results showed that calycosin enhanced cell proliferation, ALP activity and Alizarin Red-S staining in a dose-dependent manner in the range of 10(-8)-10(-6) M, while an inhibitory effect was observed at 10(-5) M. Treatment at the optimal concentration (10(-6) M, a physiologically achievable concentration) increased mRNA levels of osteogenic genes and phosphorylation of IGF1R and Akt. Furthermore, treatment with GSK or LY partly reversed the effects of calycosin on ROBs, as indicated by the decreases in calycosin-induced ALP activity, calcified nodules and osteogenic gene expression. These results suggest that the osteogenic effect of calycosin partly involves the IGF1R/PI3K/Akt signaling pathway.