酸枣仁皂苷A对小鼠MC3T3-E1成骨细胞增殖与分化的影响

Effects of jujuboside A on the proliferation and differentiation of MC3T3-E1 osteoblasts

目的通过体外细胞实验探讨酸枣仁皂苷A对小鼠MC3T3-E1成骨细胞增殖分化的影响,为临床治疗骨折与促进骨折愈合提供实验基础。方法将酸枣仁皂苷A分4个浓度(5、10、20、40 mg/L)分别干预细胞,使用CCK-8法检测细胞增殖率,流式细胞术检测细胞凋亡率,Von kossa染色法检测细胞矿化结节生成情况,Elisa法检测细胞中ALP与BGP的含量,RT-qPCR法检测细胞中Runx-2与BGP的基因表达水平,Western Blot法检测细胞中TGF-β1、Col I与BMP-2的蛋白表达水平。结果酸枣仁皂苷A能促进MC3T3-E1细胞增殖,减少细胞凋亡,提高细胞中ALP与BGP的含量,提高细胞中Runx-2与BGP基因的表达水平与TGF-β1、Col I与BMP-2的蛋白表达水平,增加细胞中矿化结节的形成数量(均P<0.05)。结论酸枣仁皂苷A能促进MC3T3-E1成骨细胞增殖,减少细胞凋亡,提高细胞分化程度与成骨效率,其机制可能是通过调控TGF/BMP通路而发挥的。

Objective To explore the effect of jujuboside A on the proliferation and differentiation of mouse MC3T3-E1 osteoblasts,and provide an experimental basis for clinical treatment of fractures and promotion of fracture healing.Methods Jujuboside A was divided into 4 concentrations(5,10,20 and 40 mg/L).Cell proliferation was detected by CCK-8,cell apoptosis was detected by flow cytometry,the cell mineralized nodule formation was detected by Von Kossa staining,the content of ALP and BGP in cells was determined by Elisa,expression levels of Runx-2 and BGP in MC3T3-E1 cells were detected by RT-qPCR,expression levels of TGF-β1,Col I and BMP-2 in MC3T3-E1 cells were detected by Western Blot.Results Jujuboside A can promote the proliferation of MC3T3-E1 cells,reduce the rate of cell apoptosis,increase the content of ALP and BGP in the cells,increase the expression levels of Runx-2 and BGP genes and the protein expression levels of TGF-β1,Col I and BMP-2,and increase the number of mineralized nodules in the cells(P<0.05).Conclusion Jujuboside A can promote the proliferation of MC3T3-E1 osteoblasts,reduce cell apoptosis,improve the degree of cell differentiation and osteogenic efficiency,and its mechanism may be exerted by regulating the TGF/BMP pathway.