P17-BMP2联合成骨诱导对大鼠骨髓间充质干细胞的影响

Effect of P17-BMP2 combined with osteogenic induction on bone marrow mesenchymal stem cells in rats

目的 探讨P17-BMP2联合成骨诱导对大鼠骨髓间充质干细胞(BMSCs)的影响。方法 选取9只3周龄Wistar雄性大鼠,分为两组培养,即A组行成骨诱导(含地塞米松7~10mg/L、维生素C 10mg/L、β-甘油磷酸钠10mmol/L的DMEM培养基培养),B组成骨诱导+P17-BMP2(成骨诱导液+P17-BMP2 10μg/ml培养),对两组大鼠进行细胞增殖、碱性磷酸酶染色(AKP)及实时荧光定量PCR技术检测,检测两组大鼠骨钙素(OCN)、骨桥蛋白(OPN)及转录因子(Runx2)的mRNA表达,以及测定两组大鼠碱性磷酸酶活性升高情况。结果 培养第5、7天后,B组BMSCs细胞增殖程度明显高于A组,差异有统计学意义(P<0.05)。诱导第7、14天,B组OPN、OCN、Runx2的mRNA表达均高于A组(P<0.05),诱导第14天,B组碱性磷酸酶活性高于A组(P<0.05)。结论 P17-BMP2联合成骨诱导能够促进骨髓间充质干细胞增殖以及成骨分化,具有较高的可研究性及操作性。

Objective To investigate the effect of P17-BMP2 combined with osteogenic induction on bone marrow mesenchymal stem cells in rats.Methods Nine 3-week-old rats were selected and divided into two groups,group A with osteogenic induction(containing dexamethasone 7~10mg/L,vitamin C 10mg/L,beta glycerin sodium phosphate tendency for 10mmol/L DMEM medium)and group B with osteogenic induction+P17-BMP2(osteogenesis induced liquid+P17-BMP210μg/ml training).Cell proliferation and alkaline phosphatase(AKP)dyeing and real-time fluorescent quantitative PCR were detected in the two groups.The mRNA expressions of OCN,OPN and Runx2,and the activity of alkaline phosphatase were detected in the two groups of rats.Results After 5 and 7 days of culture,BMSCs proliferation degree in group B was significantly higher than that in group A(P<0.05).The mRNA expressions of OPN,OCN and Runx2 on the 7th and 14th day after induction in group B were all higher than that in group A(P<0.05),and the alkaline phosphatase activity on the 14th day after induction in group B was higher than that in group A(P<0.05).Conclusion P17-BMP2 combined with osteogenic induction can promote the proliferation and osteogenic differentiation of BMSCs.