薯蓣皂苷通过信号通路对大鼠骨髓间充质干细胞成骨分化的影响  

Effect of Dioscin on Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells by Hedgehog Signaling Pathway

作　　者：颜国标 宋浩民 林南星[1] YAN Guobiao;SONG Haomin;LIN Nanxing(Department of Pharmacy,Wangjing Hospital of China Academy of Chinese Medical Sciences,Beijing 100102,China)

机构地区：[1]中国中医科学院望京医院药学部,北京100102

出　　处：《中国中医骨伤科杂志》2023年第7期1-5,共5页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：北京市科学技术委员会科研计划项目(D201300006317016)。

摘　　要：目的:探讨薯蓣皂苷对大鼠骨髓间充质干细胞(BMSCs)成骨分化及Hedgehog信号通路的影响。方法:分离培养大鼠骨髓间充质干细胞,分为对照组、薯蓣皂苷低(15%胎牛血清的DMEM培养基中加1μmol/mL薯蓣皂苷)、中(15%胎牛血清的DMEM培养基中加3μmol/mL薯蓣皂苷)、高(15%胎牛血清的DMEM培养基中加9μmol/mL薯蓣皂苷)浓度组、阳性对照组(15%胎牛血清的DMEM培养基中加成骨诱导液),培养7 d后,茜素红染色观察骨髓间充质干细胞矿化结节;细胞计数试剂盒-8法检测骨髓间充质干细胞增殖能力;检测碱性磷酸酶(ALP)水平;荧光定量PCR法检测骨髓间充质干细胞中SHH、IHH、Runx2信使RNA(mRNA)表达水平;蛋白免疫印迹法检测骨髓间充质干细胞中Smo、Gli1、Ptch1蛋白表达水平。结果:对照组可见点状矿化结节,与对照组相比,薯蓣皂苷低、中、高浓度组的矿化结节数目依次增多,体积依次增大,骨髓间充质干细胞增殖率、碱性磷酸酶水平,SHH、IHH、Runx2 mRNA,Smo、Gli1、Ptch1蛋白表达水平依次升高,差异有统计学意义(P<0.05);阳性对照组的矿化结节数目最多,连接成片,体积较大,且骨髓间充质干细胞增殖率、碱性磷酸酶水平,SHH、IHH、Runx2 mRNA,Smo、Gli1、Ptch1蛋白表达水平高于薯蓣皂苷高浓度组,差异有统计学意义(P<0.05)。结论:薯蓣皂苷可以促进大鼠骨髓间充质干细胞成骨分化,可能与Hedgehog信号通路的激活有关。Objective:To explore the effect of dioscin on osteogenic differentiation and Hedgehog signaling pathway of rat bone marrow mesenchymal stem cells(BMSCs).Methods:Rat BMSCs were isolated and cultured,and divided into control group,dioscin low(15%fetal bovine serum DMEM medium supplemented with 1μmol/mL dioscin),medium(15%fetal bovine serum DMEM medium supplemented with 3μmol/mL dioscin),high(15%fetal bovine serum DMEM medium supplemented with 9μmol/mL dioscin)concentration groups and positive control group(DMEM medium with 15%fetal bovine serum was supplemented with osteogenic induction solution).After 7 d of culture,the mineralized nodules of BMSCs were observed by Alizarin red staining,and Cell Count Kit-8 assay was used to detect the proliferation of BMSCs.Alkaline phosphatase(ALP)detection kit was used to detect ALP level.The mRNA expression levels of SHH,IHH,and Runx2 in BMSCs were detected by real-time PCR.The protein expression levels of Smo,Gli1,and Ptch1 in BMSCs were detected by Western Blot.Results:Punctate mineralized nodules were observed in the control group.Compared with the control group,the number and volume of mineralized nodules in the dioscin low,medium,high concentration groups were increased in sequence,and the proliferation rate,ALP level,SHH,IHH,Runx2 mRNA,Smo,Gli1,Ptch1 protein expression levels of BMSCs were increased in sequence(P<0.05).The positive control group had the most mineralized nodules,which connected into slices and had a large volume,and the proliferation rate,ALP level,SHH,IHH,Runx2 mRNA,Smo,Gli1,Ptch1 protein expression levels of BMSCs were higher than those in the dioscin high concentration group(P<0.05).Conclusion:Dioscin can promote the osteogenic differentiation of BMSCs in rats,which may be related to the activation of Hedgehog signaling pathway.

关 键 词：薯蓣皂苷 骨髓间充质干细胞 信号通路 成骨分化 

分 类 号：R-33[医药卫生]