葛根素对MC3T3-E1细胞增殖、分化和矿化及TRPM3 mRNA表达的影响

摘要

Aim To investigate the effect of puerarin on proliferation, differentiation, and mineralization of MC3T3-E1 cells and the expression of TRPM3 mRNA.Methods Proliferation, differentiation, and mineralization of puerarin in MC3T3-E1 cells were determined using CCK-8 assay, alkaline phosphatase(ALP) activity assay, and Alizarin Red Staining, respectively.Effect of puerarin on cell cycle and intracellular calcium concentration of MC3T3-E1 cells was detected by flow cytometry.RT-PCR was used to detect the effect of puerarin on TRPM3 mRNA expression.Resuls 0.1, 1, 10 μmol·L-1 puerarin significantly promoted the proliferation of MC3T3-E1 cells, reduced the proportion of cells in G1 phase, increased the proportion of G2 and S phase, of which 0.1 μmol·L-1 concentration effect was the most significant.Compared with control group, the ALP activity and mineralized nodule area of 0.1 μmol·L-1 puerarin group were significantly increased.The expression of TRPM3 mRNA and the intracellular calcium concentration were significantly decreased in 0.1 μmol·L-1 puerarin group.Conclusion Puerarin can promote the proliferation, differentiation, and mineralization of MC3T3-E1 cells, and reduce the expression level of TRPM3 mRNA and intracellular calcium concentration.%目的 观察葛根素对小鼠成骨细胞MC3T3-E1细胞增殖、分化和矿化及TRPM3 mRNA表达的影响.方法 分别采用CCK-8法、碱性磷酸酶(ALP)活性、茜素红染色测定葛根素对MC3T3-E1细胞增殖、成骨分化、矿化作用的影响.流式细胞术检测葛根素对MC3T3-E1细胞周期及细胞内钙离子浓度的影响.RT-PCR检测葛根素对TRPM3基因mRNA表达的影响.结果 0.1、1、10 μmol·L-1葛根素能明显促进MC3T3-E1细胞增殖,使G1期细胞比例减少,G2+S期细胞比例增加,其中0.1 μmol·L-1浓度效果最为明显;与正常对照组相比较,0.1 μmol·L-1葛根素组细胞的ALP活性和矿化结节面积均明显提高;0.1 μmol·L-1葛根素组细胞的TRPM3 mRNA表达水平和细胞内钙离子浓度明显降低.结论 葛根素可促进MC3T3-E1细胞的增殖、分化和矿化,可降低TRPM3 mRNA表达水平和细胞内钙离子浓度.