miR146a参与炎症条件下骨髓间充质干细胞成骨分化的降低及其机制研究

miR146a regulates the decrease of MSCs osteogenic differentiation in inflammation condition

目的:阐明炎症对骨髓间充质干细胞成骨分化的影响及miR146a在此过程中的调控作用。方法:将骨髓间充质干细胞置于3种不同条件培养基下培养:①成骨分化诱导培养基、②成骨分化培养基联合BMP2、③成骨分化联合炎症刺激TNF(10 ng/mL),观察其成骨分化情况。通过miRNA特异性的qRT-PCR观察miR146a在上述条件下变化情况,进而对细胞转染miR146a的拮抗体antago-miR146a观察炎症条件下骨髓间充质干细胞的成骨分化情况。结果:建立了稳定的骨髓间充质干细胞体外培养体系,在不同诱导条件下其能成骨、成脂、成软骨。BMP2能够促进MSC成骨分化,而模拟炎症刺激TNF则能抑制MSC的分化;分化条件下,miR146a表达下降;而TNF剂量依赖性能增强miR146a的表达;miR146a拮抗体则能逆转TNF引起的骨髓间充质干细胞成骨分化降低。结论:miR146a参与炎症条件下骨髓间充质干细胞成骨分化的抑制。

Objective：To investigate the role that inflammation plays in the osteogenic differentiation of MSCs and how does miR146a regulate MSCs osteogenic differentiation.Method：MSCs were cultured in 3 different condition mediums that are：①osteogenic differentiation medium which was prepared in our lab;②osteogenic differentiation medium added BMP2;③ osteogenic differentiation medium added BMP2 and TNF which mimics inflammation environment.miRNA specific qRT-PCR was applied to test miR146a level in different conditions.Antago-miR146a was transfected to test the osteogenic differentiation of MSCs in inflammation condition.Result：MSCs culture system was set up.MSC cells could differentiate into osteoblasts,condroblasts and lipoblasts.BMP2 could promote the osteogenic diffierentiation,while TNF which mimics inflammation could inhibit osteogenic diffierentiation.miR146a declined when MSCs were subjected to osteogenic medium.miR146a increased in a TNF dosage-depent way.Antago-miR146a could rescued the decrease when TNF was added.Conclusion：miR146a played an important role in inflammation induced osteogenic differentiation decrease.