中国医科大学学报2012，Vol.41Issue(7)：591-595,606,6.
miR-26a对小鼠骨髓间充质干细胞成骨分化能力的调控作用
Regulative Function of miR-26a in Enhancing the Differentiation Capacity of Bone Marrow Mesenchymal Stem Cells into Osteoblasts
摘要
Abstract
Objective To establish an osteogenic differentiation model and investigate the expression pattern and the regulative effect of miR-26a during osteogenesis. Methods BMSCs were isolated and cultured by extracting femoral bone marrow in bilateral lower limbs of female C57BL/6J mice,and were then induced into osteoblasts. The expression level of miR-26a was detected by real-time quantitative PCR in both BMSCs and osteoblasts. To investigate the function of miR-26a in BMSCs,BMSCs were transfected with miR-RiboTM microRNA-26a mimics (the augmenter of miR-26a),using siPORTTMNeoFXTM agent as the transfection reagent,and were cultured with osteogenesis inducers. Meanwhile, the control group was established. After 7 days and 14 days of culture, the cells were collected and examined by cell morphology, alkaline phosphatase staining,calcium nodules (Alizarin red staining) and so on. In vivo expression of tniR-26iL was detected by real-time PCR in the BMSCs of both the animal postmenopausal osteoporosis models (OVX group) and the sham group. Results There was an approximately 25-fold increase of miR-26a expression in BMSCs during osteogenesis. Meanwhile, the expression of osteogenetic genes including 0CN,Runx-2 and Col-1 also increased following a time-dependent pattern during the osteogenetic induction. BMSCs transfected with miR-26a mimics changed from the fusiform shape into the polygonal shape after being cultured for 7 days,which was the same as the control group. ALP staining revealed that after 7 days, the ALP activity of BMSCs transfected with miR-26a mimics was higher than that of BMSC transfected with mimics control. Alizarin red staining showed tbat calcium deposits were more and bigger in experiment group compared with control group. The PCR showed that the expression of osteogenetic genes ( OCN, Runx-2 and Col-1) were higher in miR-26a mimics-transfecting group. The expression of miR-26a in vivo was 4 times lower in OVX group than in sham group. Conclusion miR-26a mimics can enhance os-teogenesis of BMSCs, which may confirm the potential role of miR-26a in promoting the osteogenetic capacity of BMSCs.关键词
miRNA-26a/成骨分化/骨髓间充质干细胞Key words
miRNA-26a/osteogenetic differentiation/bone marrow mesenchymal stem cells分类
医药卫生引用本文复制引用
范龙坤,华泽权,金岩,时炳正..miR-26a对小鼠骨髓间充质干细胞成骨分化能力的调控作用[J].中国医科大学学报,2012,41(7):591-595,606,6.基金项目
国家基础研究项目(973项目)(2011CB964700) （973项目）
