miR-21对绝经后骨质疏松骨髓间充质干细胞成骨能力影响的研究

摘要

目的 探讨miR-21对绝经后骨质疏松骨髓间充质干细胞(PMOP-hBMMSC)成骨能力影响.方法 分离培养正常人(H-hBMMSCs)和绝经后骨质疏松患者(PMOP-hBMMSCs)的骨髓间充质干细胞,并对两组细胞的成骨能力进行比较.转染上调PMOP-hBMMSCs中miR-21表达,观察分析其对成骨能力的影响.Real time RT-PCR和Western Blot比较miR-21在三组细胞间的表达差异,同时检测三组成骨标志性基因Runx2和Osterix的表达.结果 PMOP-hBMMSCs组中的miR-21表达水平、ALP的活性以及钙结节染色面积都高于PMOP组(P＜0.05),与H-hBMMSCs正常对照组差异无统计学意义(P＞0.05);RT-PCR和Western Blot结果显示成骨诱导7d后,转染后高表达Runx2和Osterix mRNA和蛋白水平均高于PMOP组(P＜0.05,P＜0.01),与H-hBMMSCs正常对照组差异无统计学意义(P＞0.05).结论 转染miR-21后的PMOP-hBMMSCs成骨能力增强.%Objective To investigate the osteogenic capability of miR-21 in human bone marrow mesenchymal stem cells in patients with postmenopausal osteoporosis (PMOP-hBMMSCs).Methods The human bone marrow mesenchymal stem cells (hBMMSCs) and PMOP-hBMMSCs were isolated and cultured.The osteogenic capabilities of these two kinds of cells were compared.After over-expression miR-21,the osteogenic potential was analysed in PMOP-hBMMSCs.The expression of Runx2 and Osterix,genes in H-hBMMSCS and PMOP-hBMMSCs were respectively detected by real time RT-PCR and western blot.Results The expression level of miR-21 was higher,ALP was more active,and the calcium nodules staining area was larger in the PMOP-hBMMSCs group than in the PMOP group(P ＜ 0.05).Up-regulation was found in the expressions of Runx2 and Osterix mRNA and protein of Over-expression of miR-21 PMOP-hBMMSCs compared to PMOP(P ＜ 0.05,P ＜ 0.01).Conclusion After the transfection of miR-21,the osteogenic ability of PMOP-hBMMSCs can be enhanced.