珠子参提取物促进大鼠成骨细胞增殖、分化、矿化及调节OPG/RANKL表达的作用

Effect of Panax japonicas extract on the proliferation, differentiation, mineralization and expression of OPG/RANKL in rat osteoblasts

目的探讨珠子参对原代大鼠成骨细胞增殖、分化、矿化及对骨保护蛋白(OPG)和核因子-κβ受体活化因子配体(RANKL)表达的影响。方法离体培养大鼠原代成骨细胞;采用MTT法、Ed U染色法、测定碱性磷酸酶法、矿化结节染色法考察不同浓度珠子参提取物对成骨细胞活力、增殖、分化和矿化等作用的影响;采用Real time PCR法和Western blot法检测珠子参提取物对成骨细胞OPG及RANKL mRNA和蛋白表达的调节作用。结果珠子参提取物浓度在500μg·mL^-1时可提高成骨细胞活力;在300、400和500μg·mL^-1时可显著提高成骨细胞增殖、分化和矿化能力,提高OPG mRNA和蛋白的表达,降低RANKL mRNA和蛋白表达,且呈现剂量依赖性。结论珠子参提取物可在一定浓度范围内提高大鼠成骨细胞的活力、增殖、分化和矿化,同时可调节OPG/RANKL表达,这可能是其防治骨质疏松症的作用机制。

Objective To determine the effect of Panax japonicas extract on the proliferation,differentiation,mineralization and mRNA expression of osteoprotegerin(OPG)and receptor activator of nuclear factor kappa B ligand(RANKL)in primary rat osteoblasts.Methods Rat primary osteoblasts were cultured in vitro.The effect of Panax japonicas extract on the osteoblasts viability,proliferation,differentiation and mineralization was measured by MTT,EdU staining,alkaline phosphatase assay and mineralized nodule staining.The effect of Panax japonicas extract on the mRNA expression of OPG and RANKL in osteoblasts was measured by Real time PCR,and the protein expression measureel by Western blot.Results The activity of osteoblasts was significantly increased by the extract of Panax japonicas at 500μg·mL^-1.The proliferation,differentiation and mineralization of osteoblasts were significantly increased at 300,400 and 500μg·mL^-1.The mRNA and protein expression of OPG was significantly increased.However,RANKL mRNA was decreased by the extract of Panax japonicas in a dose-dependent manner.Conclusion The extract of Panax japonicas can increase the viability,proliferation,differentiation and mineralization of rat osteoblasts,and regulate the expression ratio of OPG/RANKL at appropriate concentrations.This may be the basis of Panax japonicas in preventing osteoporosis.