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

Effect of puerarin on proliferation,differentiation and mineralization of MC3T3-E1 cells and expression of TRPM3 mRNA

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m RNA表达的影响。结果 0.1、1、10μmol·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,G_2+S期细胞比例增加,其中0.1μmol·L^(-1)浓度效果最为明显;与正常对照组相比较,0.1μmol·L^(-1)葛根素组细胞的ALP活性和矿化结节面积均明显提高;0.1μmol·L^(-1)葛根素组细胞的TRPM3 m RNA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。

Aim To investigate the effect of puerarin on proliferation,differentiation,and mineralization of MC3T3-E1 cells and the expression of TRPM3 mR NA.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 mR NA expression.Resuls0.1,1,10 μmol·L^-1puerarin 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^-1concentration effect was the most significant.Compared with control group,the ALP activity and mineralized nodule area of 0.1 μmol · L^-1puerarin group were significantly increased.The expression of TRPM3 mR NA and the intracellular calcium concentration were significantly decreased in 0.1 μmol · L^-1puerarin group.Conclusion Puerarin can promote the proliferation,differentiation,and mineralization of MC3T3-E1 cells,and reduce the expression level of TRPM3 mR NA and intracellular calcium concentration.