二甲双胍改善长期体外培养骨髓间充干细胞的增殖和分化功能

The effects of metformin on the proliferation and osteogenic differentiation of long- term cultured bone marrow mesenchymal stem cells

目的:观察二甲双胍(metfomin, Met)能否改善体外长期培养小鼠骨髓间充质干细胞(BMMSCs)的增殖和成骨分化功能。方法:分离小鼠BMMSCs进行体外培养,选取P1代和P6代细胞,P6代细胞用Met处理,检测细胞增殖,克隆形成,ALP活性、矿化结节和成骨基因mRNA表达水平。使用试剂盒检测细胞ROS水平,并检测抗氧化、干细胞属性(干性)及衰老相关基因的mRNA表达情况。结果:小鼠P6代BMMSCs的增殖和成骨分化功能明显低于P1代细胞,Met可以显著提高P6代细胞的增殖和成骨分化能力,并提高细胞的抗氧化功能和干性,降低衰老相关基因表达。结论:Met能够增强体外长期培养的小鼠BMMSCs的增殖和分化功能,这种作用可能是通过改善氧化应激和提高干性实现的。

Objective: To investigate the effects of metformin(Met) on the proliferation and osteogenic differentiation of long-term cultured bone marrow mesenchymal stem cells(BMMSCs). Methods: BMMSCs were harvested from C57 mice and cultured in vitro. The 6 th passaged BMMSCs were treated with Met. The proliferation, colony formation, ALP activity, mineralization and mRNA levels of osteogenic genes were examined. ROS content and mRNA levels of antioxidant, stemness and senescence genes were detected. Results: The proliferation and osteogenic differentiation ability of the 6 th passaged BMMSCs were significantly lower than those of the 1 st passaged BMMSCs, which could be rescued by Met. The antioxidant ability and stemness of the 6 th passaged BMMSCs were improved by Met while the senescence related genes were suppressed by Met. Conclusion: Metformin can improve the proliferation and differentiation ability of long-term cultured mice BMMSCs, which may be mediated via alleviating oxidative stress and promoting stemness.