淫羊藿苷对体外在无地塞米松培养基中诱导培养大鼠骨髓基质细胞成骨性分化的影响

Icariin induces osteogenic differentiation of bone marrow stromal cells in the absence of dexamethasone in vitro

目的探讨在无地塞米松(dexamethasone)时淫羊藿苷(icariin,ICA)对体外培养大鼠骨髓基质细胞成骨性分化的影响,并试图寻找一种新的或更强的促体外培养骨髓基质细胞成骨性分化的诱导剂。方法用1×10-5mol·L-1淫羊藿苷或1×10-8mol·L-1的地塞米松对大鼠骨髓基质细胞分别进行药物干预,细胞计数试剂盒(Cell Counting Kit-8,CCK-8)检测细胞增殖,碱性磷酸酶检测试剂盒检测碱性磷酸酶(alkaline phosphatase,ALP)活性,茜素红染色检测钙化结节数量,实时定量聚合酶链反应(RT-PCR)检测骨保护素(osteoprotegerin,OPG)、核因子-κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)、骨形态发生蛋白(bone morphogenetic protein,BMP)的信使RNA(mRNA)表达情况。结果 1×10-5mol·L-1淫羊藿苷在无地塞米松的培养基中能显著提高大鼠骨髓基质细胞ALP活性;明显增加大鼠骨髓基质细胞钙化结节数目;显著上调BMP、OPG mRNA水平以及OPG/RANKL mRNA比值。结论淫羊藿苷可能是一种潜在的促体外培养大鼠骨髓基质细胞成骨性分化的新诱导剂。

Objective To investigate the role of icariin in inducing osteogenic differentiation of bone marrow stromal cells without dexa-methasone,to seek a novel and stronger inducer for promoting osteogenic differentiation of bone marrow stromal cells in vitro.Methods The Bone marrow stromal cells were cultured in DMEMcontaining 1 ×10 -5 mol·L -1 icariin or 1×10 -8 mol·L -1 dexamethasone. Cell proliferation was determined using CCK-8 kit;ALP activity of the culture was measured using commercial kit of ALP;and calcified nodule was stained with alizarin red.The mRNA levels of OPG,RANKL and BMP were measured by RT-PCR.Results Without dexa-methasone,icariin could in vitro prominently promote ALP activity of the bone marrow stromal cells,and up-regulate the mRNA levels of OPG and BMP,as well as OPG/RANKL mRNA ratio.Conclusions Icariin could act as a novel inducer in promoting osteogenic differ-entiation of bone marrow stromal cells in vitro.