松果菊苷对大鼠成骨细胞胃桥素基因和蛋白质的影响

为了探讨松果菊苷对体外培养大鼠成骨细胞骨桥素(OPN)mRNA和蛋白质表达的影响,选取出生24 h内的SD大鼠颅盖骨,依次用胰酶和胶原酶消化分离培养成骨细胞,用不同浓度的松果菊苷作用于第3代细胞,选取雌二醇为阳性对照,分别在药物作用24、48和72 h后提取细胞总RNA的表达量,用荧光定量PCR(FQ-PCR)法检测细胞中OPN mRNA的表达量;用Western blotting检测细胞中OPN蛋白的表达量。结果显示,各浓度松果菊苷在各个时间段均增加OPN基因的表达,尤其是5×10^-5和5×10^-6 mg/mL松果菊苷在各时间段均具有显著或极显著地促进作用(P＜0.05或P＜0.01);各浓度组在作用48 h时蛋白质的表达量均高于对照组。结果表明,松果菊苷对成骨细胞OPN基因表达有显著的促进作用,同时对OPN蛋白的分泌有促进的趋势。

Effect of Echinacoside on the Expression of the OPN mRNA and Protein in Rat Osteoblasts in vitro

In order to investigate the effect of Echinacoside on the OPN mRNA and protein expression of rat osteoblasts cultured in vitro, the osteoblasts were derived from calvariae of neonatal rat and digested by the trypsin and collagenase. The third-generation of cultured osteoblasts were exposed to different concentrations of Echinacoside, while took oestrogen as positive control. Total RNA and protein were prepared from osteoblasts after 24, 48 and 72 h respectively. And the change from OPN mRNA was dected by fluorescent quantitation PCR,OPN protein was dected by Western blotting. The results indicated that different concentrations of Echinacoside had increased the expression of osteoblast OPN mRNA, especially 5?10^-4 and 5?10^-5 mg/mL Echinacoside significantly or extremly significantly up-regulated the expression of OPN mRNA at 24, 48 and 72 h. Secretion of OPN protein was more than control group after 48 h addition of Echinacoside.The conclusion was that Echinacoside significantly promoted the expression of OPN mRNA of osteoblasts, while had the trend of raised role for OPN protein.