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

作　　者：曾祥伟[1] 冯倩[1] 张莹莹[2] 赵凤鸣[1] 詹秀琴[1] 

机构地区：[1]南京中医药大学医学与生命科学院,江苏南京210023 [2]南京医科大学第二附属医院疼痛科,江苏南京210011

出　　处：《中国药理学通报》2017年第7期977-982,共6页Chinese Pharmacological Bulletin

基　　金：江苏省自然科学基金项目(No BK20131417);江苏高校优势学科建设工程资助项目(中西医结合)

摘　　要：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.

关 键 词：葛根素 MC3T3-E1细胞 增殖 分化 矿化 TRPM3 

分 类 号：R-332[医药卫生] R284.1