基于Toll样受体4通路研究黄芪甲苷促进成骨细胞增殖分化的作用

The effect of astragaloside Ⅳ on proliferation and differentiation of osteoblast on the basis of TLR-4 pathway

目的研究黄芪甲苷促进成骨细胞增殖分化的作用,并初步探讨作用机制。方法将成骨细胞MC3T3-E1分为对照组和实验组,对照组正常培养成骨细胞,实验组采用不同浓度黄芪甲苷干预成骨细胞,并孵育不同时间,MTT法观察对细胞增殖的影响,检测碱性磷酸酶(ALP)活性观察对细胞分化的作用,Western blot检测成骨细胞Toll样受体-4(TLR-4)蛋白表达。结果与对照组比较,随着黄芪甲苷浓度的增加,成骨细胞活性或ALP活性逐渐上升,并趋于稳定。延长黄芪甲苷作用时间,低浓度组细胞活性显著升高,而各组细胞ALP活性均显著上升。与对照组比较,实验组细胞中TLR-4表达显著下降。结论黄芪甲苷能够促进成骨细胞增殖和分化,其作用机制可能与TLR-4通路有关。

Objective To study the effect of astragaloside Ⅳ on proliferation and differentiation of osteoblast,and preliminarily explore the mechanism of action. Methods The osteoblast MC3T3-E1 cell was divided into control group and experimental group. The osteoblast of control group was cultured normally,while the one of experimental group was co-cultured with astragalosideⅣ of different concentrations for different time. The MTT assay was used to observe the proliferative effect,and the activation of alkaline phosphatase(ALP)was used as an index to evaluate the differentiation level of cells. The expression of toll-like receptor-4(TLR-4)was detected by Western blot. Results With the increase of astragaloside Ⅳ concentration or the incubating time,the ability of osteoblast proliferation or differentiation gradually,which became stable. With the increase of time,the cell activation of low concentration groups increased significantly,and the ALP activation of all groups increased significantly. Compared with the control group,the expression of TLR-4 in the experimental group was significantly decreased. Conclusion Astragaloside Ⅳ could promote the proliferation and differentiation of osteoblast,which may involved with the TLR-4 pathway.