骨髓间充质干细胞体外分化过程中复方接骨片和壮骨片含药血清的作用

Effect of Compound Jiegu Tablets and Zhuanggu Tablets on the differentiation of bone marrow mesenchymal stem cells in vitro

背景:骨髓间充质干细胞是存在于骨髓中的一种具有多向分化潜能的干细胞,当受到含药血清作用时,其所涉及的多条信号通路上的基因表达会发生改变。目的:考察复方接骨片和壮骨片含药血清在骨髓间充质干细胞体外分化过程中的作用。方法:取60只SD大鼠,随机分为复方接骨片组、壮骨片组、两药联用组和空白对照组,制备含药血清。分离培养SD大鼠骨髓间充质干细胞,于培养瓶中加入不同的含药血清,培养24 h后,RT-PCR法检测BMP-2、Runx2、VEGF及Akt m RNA的表达,Western blot法检测BMP-2、Runx2、VEGF及Akt蛋白的表达。结果与结论:(1)与对照组比较,复方接骨片组及两药联用组Akt和VEGF m RNA及蛋白的表达明显升高(P<0.05或P<0.01);(2)壮骨片组与对照组相比,Akt和VEGF m RNA及蛋白表达差异无显著性意义(P>0.05);(3)与对照组比较,复方接骨片组BMP-2和Runx2 m RNA及蛋白表达升高(P<0.05);(4)与对照组相比,壮骨片组及两药联用组BMP-2及Runx2 m RNA及蛋白表达均显著增加(P<0.01);(5)结果表明,复方接骨片及壮骨片含药血清可在骨髓间充质干细胞体外分化通路中起到协同促进骨髓间充质干细胞向血管内皮细胞及成骨细胞分化的作用。

BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)are a kind of pluripotent stem cells in the bone marrow.Drug-containing serum treatment may be followed by a variation in the expression of genes involved in multiple signaling pathways.OBJECTIVE:To study the effect of compound Jiegu Tablets and Zhuanggu Tablets on the differentiation of BMSCs in vitro.METHODS:Sixty Sprague-Dawley rats were randomized into Jiegu group,Zhuanggu group,combined group and blank control group.Different drug-containing sera were prepared and used to culture BMSCs isolated from the rat bone marrow.RT-PCR and western blot assay were used to detect the expression of BMP-2,Runx2,VEGF and Akt at mRNA and protein levels,respectively,at24hours after culture.RESULTS AND CONCLUSION:Compared with the blank control group,the expression levels of Akt and VEGF mRNA and protein were significantly increased in the Jiegu group and combined group(P<0.05or P<0.01),but showed no difference in the Zhuanggu group(P>0.05).Compared with the blank control group,the expression levels of BMP-2and Runx2mRNA and protein were significantly increased in the Jiegu group(P<0.05)and very significantly increased in the Zhuanggu group and combined group(P<0.01).These findings indicate that compound Jiegu Tablets and Zhuanggu Tablets containing sera lead to synergistic effects on the BMSCs differentiation into vascular endothelial cells and osteoblasts in vitro.