目的 观察淫羊藿次苷Ⅰ (Icariside Ⅰ)与其原型药物淫羊藿苷(Icariin,ICA)对大鼠骨髓间充质干细胞(rat bone marrow stromal cells,rBMSCs)成骨分化过程中碱性磷酸酶(ALP)、骨钙素(BGP)蛋白表达量及ALP、BGP、Osterix和Runx-2 mRNA表达的影响.方法 贴壁筛选法体外培养rBMSCs,以1×10-5 mol/L的ICA和Icariside Ⅰ对rBMSCs的成骨性分化进行药物干预.ELISA法检测ICA组、Icariside Ⅰ组和空白对照组及转化液组之间ALP活性、BGP分泌量;RT Real-Time PCR法检测ALP、BGP、Osterix和Runx-2 mRNA基因表达.结果 与空白对照组相比,Icariside Ⅰ与其原型药物ICA均可显著增强rBMSCs的ALP活性,促进BGP的分泌,提高ALP、BGP、Osterix和Runx-2的mRNA水平(P＜0.01).Icariside Ⅰ与ICA组比较,其ALP、BGP蛋白表达量及基因表达明显高于ICA组(P＜0.01),Osterix和Runx-2的基因表达两组无显著性差异.结论 Icariside Ⅰ 也能促进rBMSCs成骨性分化,并且其活性高于原型药物ICA,也是淫羊藿发挥抗骨质疏松活性的主要成分.②提示补肾中药淫羊藿其补肾壮骨作用与促进ALP、BGP蛋白分泌量,上调ALP、BGP、Osterix和Runx-2的mRNA水平有关.%Objective To evaluate the effects of icariin and its major metabolite Icariside Ⅰ on the protein expression of ALP and BGP and the mRNA expression of ALP,BGP,Osterix and Runx-2 during osteogenic differentiation in rat bone marrow stromal cells (rBMSCs).Methods By using the adherencemethod,BMSCs were cultured.Icariin and Icariside Ⅰ were supplemented into the culture at 1 × 10-5 mol/L,respectively.The experiment groups were blank control,positive control,Icariin and Icariside Ⅰ groups.ELISA was performed to examine the protein expression of ALP and BGP on the 62,9th,and 12th day.The expressions of ALP,BGP,Osterix and Runx-2 genes were detected using RT Real-Time PCR.Results Compared with that of the blank control group,Icariin and Icariside Ⅰ significantly improved ALP activity and ALP and BGP amount (P ＜ 0.01),and enhanced the gene expressions of ALP,BGP,Osterix and Runx-2 (P ＜ 0.01).Compared with the Icariin group,Icariside I had significantly stronger effect on the gene expressions of ALP and BGP protein (P ＜0.01),but there were no significant differences between the groups for Osterix and Runx-2 gene expressions.Conclusion 1) Icariside could promote the osteogenic differentiation of rBMSCs greater than Icariin,and is the main active anti-osteoporosis ingredient of the herb epimedii;2) The function of the herb epimedii in strengthening bone of is associated with the increased production of ALP and BGP,and the upregulation of the mRNA expression of ALP,BGP,Osterix and Runx-2.
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