松果菊苷含药血清促进大鼠骨髓间充质干细胞成骨分化及ZHX3表达的影响

Effect of echinacoside-containing serum in promoting mesenchymal stem cell osteogenic differentiation and ZHX3expression in rats

探讨松果菊苷含药血清促进大鼠骨髓间充质干细胞向成骨细胞分化的作用及可能的机制。采用全骨髓贴壁法培养大鼠骨髓间充质干细胞,取第3代细胞分为3组:空白对照组、经典成骨诱导组、10%松果菊苷含药血清组。ELISA测定碱性磷酸酶和骨钙素的表达;免疫荧光和Western blot检测ZHX3蛋白的表达,RT-PCR检测ZHX3mRNA的表达。结果显示,经典成骨诱导组和10%松果菊苷含药血清组的碱性磷酸酶和骨钙素表达明显增强,与空白对照组比较有显著性差异(P<0.01);10%松果菊苷含药血清组的碱性磷酸酶和骨钙素表达与经典成骨诱导组比较明显增强,有显著性差异(P<0.01)。经典成骨诱导组和10%松果菊苷含药血清组的ZHX3蛋白及mRNA表达明显增强,与空白对照组比较有显著性差异(P<0.01);10%松果菊苷含药血清组的ZHX3蛋白及mRNA表达与经典成骨诱导组比较明显增强,有显著性差异(P<0.01)。10%松果菊苷含药血清可以促进体外培养的骨髓间充质干细胞向成骨细胞分化,其作用机制与ZHX3表达增加有关。

To investigate the effect and possible mechanism of echinacoside-containing serum on the osteogenic differentiation in rat bone marrow mesenchymal stem cells. Rat bone marrow mesenchymal stem cells were cultivated by the whole bone marrow adherence method. The 3rdgeneration of cells were divided into 3 groups： the blank control group,the classic osteogenic-induced group and the10% echinacoside-containing serum group. The expression of alkaline phosphatase and osteocalcin were detected by ELISA. The expression of ZHX3 protein was detected by Western blot technique. RT-PCR technique was used to detect the expression of ZHX3 mRNA. According to the result,the expressions of the alkaline phosphatase and osteocalcin in the classic osteogenic-induced group and the 10% echinacoside-containing serum group were significantly higher than that of the blank control group（ P〈0. 01）. And expressions of the alkaline phosphatase activity and osteocalcin in the 10% echinacoside-containing serum group were significantly higher than that in the classic osteogenic-induced group（ P〈0. 01）. Meanwhile,the classic osteogenic-induced group and the 10% echinacosidecontaining serum group showed obviously higher ZHX3 protain and mRNA expression than that of the black control group,with significant differences（ P〈0. 01）; the 10% echinacoside-containing serum group showed obviously higher ZHX3 protain and mRNA expression than that of the classic osteogenic-induced group,with a significant difference（ P〈0. 01）. In conclusion,10% echinacoside-containing serum can promote the differentiation of the bone marrow mesenchymal stem cells cultured in vitro. Its mechanism may be correlated with the increase in the ZHX3 expression.