摘要
目的:研究淫羊藿苷含药血清(SI)对体外培养大鼠骨髓间充质干细胞(rBMSCs)增殖和成骨性分化的影响。方法:以每0.25g/kg质量淫羊藿苷的剂量灌服成年大鼠,制得淫羊藿苷含药血清,以灌服等体积生理盐水制得对照血清。分别以2.5%,5%和10%3种血清浓度干预rBMSCs,MTT法检测细胞增殖率,同时检测成骨性诱导培养9d后的碱性磷酸酶(ALP)活性,筛选出最佳血清浓度。以最佳血清浓度干预rBMSCs的成骨性分化,比较SI组和对照组之间的钙盐沉积量、骨钙素分泌量;提取Total RNA,RT-PCR检测IGF-1,Runx-2与Osterix mRNA的表达情况。结果:所有浓度的含药血清均能抑制rBMSCs的增殖,但可提高ALP活性。5%的SI浓度为最佳干预浓度,该浓度的含药血清可显著促进骨钙素的分泌和钙盐沉积,提高ALP活性,增强IGF-1,Runx-2与Osterix mRNA的表达。结论:淫羊藿苷经口服后的代谢产物抑制rBMSCs增殖,但可促进成骨性分化,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 the serum of rats administered icariin（SI） on the proliferation and osteogenic differentiation of rat bone marrow stromal cells（rBMSCs）.Methods： Adult Wistar rats were given icariin intragastrically at 250mg/kg body weight and their serum was obtained after 1 hour.The control serum was obtained by giving equal volume of physiological saline.The serums were supplemented into the culture media of rBMSCs at 2.5%,5% and 10% respectively.MTT method was adopted in proliferation analysis.The activity of ALP was examined after induced 9 days.Search the best serum concentration and supplemented into the culture medium,then the osteogenic differentiation markers including osteocalcin secretion,calcium deposition were compared between the SI-supplemented group and the control.Total RNA was isolated and the gene expression of IGF-1,Runx-2 and Osterix were investigated by Real Time RT-PCR.Results： All the concentration of SI can refrain the proliferation of rBMSCs,but it evidently promotes the activity of ALP.5% is the best concentration.This concentration can significantly improve ALP activity,osteocalcin secretion and calcium deposition.Besides,it also can enhance the mRNA level of IGF-1,Runx-2 and Osterix.Conclusion： The metabolites of icariin after oral administration inhibit the proliferation of rBMSCs,but stimulate its osteogenic differentiation,suggesting that icariin can be administered via oral and its metabolites are the effective constitutes for anti-osteoporosis activity.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2012年第2期480-484,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
甘肃省科技计划资助项目(No.092NKDA025)~~
