淫羊藿总黄酮含药血清对成骨细胞OPG/OPGL基因表达的影响

Effects of Drug-containing Serum of Total Flavone of Epimedium on Expression of OPG/OPGL Gene Osteoblasts

目的研究淫羊藿总黄酮含药血清对体外培养成骨细胞骨保护素(OPG)和骨保护素配体(OPGL)基因表达的影响,进一步揭示淫羊藿"补肾壮阳,益精健骨"的作用机制。方法采用中药血清药理学方法制备含淫羊藿总黄酮大鼠血清;用多次酶消化法体外培养成骨细胞;将含淫羊藿总黄酮大鼠血清(SRAT)以5%的质量浓度加入新生大鼠颅骨成骨细胞培养液中,研究它们对OPG/OPGL基因表达水平的影响。OPG/OPGL基因表达水平用RT-PCR法检测。结果半定量RT-PCR显示5%含淫羊藿总黄酮大鼠血清干预成骨细胞时,存在OPG mRNA的转录,在此浓度下,随着时间的延长转录水平逐渐增加,而OPGL mRNA水平随着时间的延长转录水平逐渐减弱。结论淫羊藿总黄酮代谢产物能上调成骨细胞中OPG mRNA的表达、抑制OPGL mRNA的表达,说明淫羊藿总黄酮代谢产物具有促进骨形成活性。

ObjectiveTo investigate the effects of drug-containing serum of total flavone of Epimedium on the gene expression of osteoprotegerin and osteoprotegerin ligand of rat osteoblasts in vitro,in order to understand its mechanism of ＂strengthening kidney-yang and bone＂. MethodsThe rat calvarial osteoblasts（ROB） were released from newborn calvarial bone pieces after repeatedly digested with the mixture of collagenase and trypsin,and fed with the culture medium α-MEM.The serum of rats administered total flavone of Epimedium was obtained by the method of serum pharmacology.The serum was supplemented into the culture media by 5% respectively.OPG/OPGL gene expression was assayed by RT-PCR. ResultsRT-PCR showed that the expression level of OPG was increased when 5% SRAT was supplemented,while the level of OPGL was reduced. ConclusionThe metabolites of the total flavone of Epimedium after oral administration strongly stimulate the proliferation and osteogenic differentiation of osteoblasts and regulate the value of OPG/OPGL,which indicates that they are the effective constituentes of bone formation.