目的 研究二甲双胍对PMMA颗粒刺激下人间充质干细胞成骨分化的影响.方法 体外培养人胎盘间充质干细胞,CCK8法检测二甲双胍对细胞活力的影响;实时定量PCR分析成骨相关基因OCN,RNUX2及ALP表达;茜素红染色分析钙沉积状况;实时定量PCR分析eNOS表达.结果 PMMA能抑制人间充质干细胞活性,0.05 mmol/L二甲双胍能缓解PMMA对细胞活性的抑制;PMMA能抑制间充质干细胞成骨相关基因表达及矿物质沉积,加入二甲双胍后成骨基因表达升高,钙沉积增强;二甲双胍能上调间充质干细胞eNOS表达.结论 二甲双胍能有效促进PMMA颗粒作用下的间充质干细胞成骨分化,其机制可能与上调eNOS表达有关.%Objective To investigate the effect of metformin on the osteogenic differentiation of human mesenchymal stem cells exposed to PMMA particles. Methods Human placental mesenchymal stem cells were iso-lated and cultured in vitro. The effect of metformin with different concentrations on cell viability was determined by CCK8 assay. The effect of metformin on the mRNA expression of osteogenic genes was detected by using real-time RT-PCR. Calcified nodules were stained by alizarin S. The effect of metformin on the expression of eNOS was also detected by using real-time RT-PCR. Results PMMA particles could inhibit the viability of mesenchymal stem cells. Metformin(0.05 mmol/L)could promote the viability of mesenchymal stem cells exposed to PMMA particles. Metformin(0.05 mmol/L)could increase the expression of osteogenic genes,including OCN,RNUX2,and ALP, in human mesenchymal stem cells exposed to PMMA particles. The calcium deposit was also increased after metfor-min treatment. Results of real-time RT-PCR showed that metformin could increase the expression of eNOS in human mesenchymal stem cells exposed to PMMA particles. Conclusions Metformin can increase the osteogenic differentiation of human mesenchymal stem cells exposed to PMMA particles,partially by inducing eNOS expression.
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