淫羊藿苷与其代谢产物淫羊藿次苷Ⅱ对骨髓间充质干细胞成骨性分化影响的比较研究

Comparative Study on the Osteogenic Differentiation of Rat Bone Marrow Stromal Cells Effected by Icariin and IcarisideⅡ

目的:研究淫羊藿苷与其主要代谢产物——淫羊藿次苷Ⅱ对体外培养大鼠骨髓间充质干细胞(Rat bonemarrow stromal cells,rBMSCs)成骨性分化的影响。方法:贴壁筛选法体外培养rBMSCs,以5×10-5mol/L的淫羊藿苷和淫羊藿次苷Ⅱ对rBMSCs的成骨性分化进行药物干预,比较淫羊藿苷组、淫羊藿次苷Ⅱ组和不加药的对照组之间碱性磷酸酶(ALP)活性、碱性磷酸酶阳性克隆数(CFU-FALP)、骨钙素分泌量、钙盐沉积量以及钙化结节数量的差异,RTReal-Time PCR法检测bFGF、IGF-1、Osterix和Runx-2的基因表达情况。结果:淫羊藿苷和淫羊藿次苷Ⅱ均可显著增强rBMSCs的ALP活性,促进钙盐沉积和骨钙素的分泌,增加CFU-FALP和钙化结节数量,提高bFGF、IGF-1、Osterix和Runx-2的mRNA水平,但淫羊藿次苷Ⅱ的活性明显高于淫羊藿苷。结论:淫羊藿次苷Ⅱ促进rBMSCs成骨性分化的活性高于淫羊藿苷,提示淫羊藿苷可以通过口服途径给药,其代谢物是发挥抗骨质疏松活性的主要成分。

Objective：To investigate the effects of icariin and it′s main metabolites-icariside Ⅱ on the osteogenic differentiation of rat bone marrow stromal cells（rBMSCs）.Methods：rBMSCs were cultured by adherence screening method,icariin and icariside Ⅱ were supplemented into the culture at 5×10-5mol/L respectively.The osteogenic differentiation markers including alkaline phosphatase（ALP） activity,CFU-FALP,osteocalcin secretion,calcium deposition and mineralized bone modulus were compared among the icariin-supplemented group,icariside Ⅱ and the control.The gene expressions of bFGF,IGF-1,Osterix and Runx-2 were examined by RT-Real Time PCR.Results：Both icariside Ⅱ and icariin significantly improved ALP activity,CFU-FALP amount,osteocalcin secretion,calcium deposition and mineralized modulus.Besides,they enhanced the gene expressions of bFGF,IGF-1,Osterix and Runx-2.Icariside Ⅱ was obviously stronger than icariin at the above activities.Conclusion：Icariside Ⅱ is stronger than icariin at enhancing the osteogenic differentiation of rBMSCs,suggesting that icariin can be administered via oral and it′s metabolites are the effective constitutes for antiosteoporosis activity.