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目的:探索对骨髓间充质干细胞(BMMSCs)具有最佳促成骨能力的葛根素浓度以及与Wnt信号通路之间的相关性。方法:取3周龄的SD大鼠,分离培养BMMSCs,传至第3代后接种,改用含不同浓度葛根素的培养基。继续培养2~4周,测定碱性磷酸酶与骨钙素的含量,评价葛根素的体外诱导成骨能力。用Elisa试剂盒测定Wnt5b的含量,评估葛根素的促成骨作用与Wnt信号通路的关系。采用SPSS16.0软件包对数据进行统计学分析。结果:不同浓度组的细胞ALP、OCN和Wnt5b含量增加,10~(-5) mol/L组显著高于对照组,高于其他浓度组,但差异无显著性。结论 :葛根素能有效促进BMMSCs向成骨方向分化,以10~(-5) mol/L浓度效果最佳,其作用机制可能与Wnt信号通路有关。
OBJECTIVE: To explore the correlation between the concentration of puerarin with Wnt signaling pathway and the best osteogenic potential of bone marrow mesenchymal stem cells (BMMSCs). Methods: Three-week-old SD rats were used to separate and culture BMMSCs. After passage 3, they were inoculated and replaced with medium containing different concentrations of puerarin. Continue to train 2 to 4 weeks, the determination of alkaline phosphatase and osteocalcin content, evaluation of puerarin in vitro induced osteogenesis. The Wnt5b content was determined by Elisa kit and the relationship between puerarin-induced bone formation and Wnt signaling pathway was evaluated. SPSS16.0 software package for statistical analysis of the data. Results: The contents of ALP, OCN and Wnt5b increased in different concentration groups, while the levels in 10 -5 mol / L group were significantly higher than those in control group, but not significantly different. CONCLUSION: Puerarin can effectively promote BMMSCs to differentiate into osteoblasts. The optimal concentration of 10 ~ (-5) mol / L puerarin is the best, and its mechanism may be related to Wnt signaling pathway.