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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 elucidate the effect of inflammation on osteogenic differentiation of bone marrow mesenchymal stem cells and the role of miR146a in this process. Methods: Bone marrow mesenchymal stem cells were cultured under three different conditions: osteogenic differentiation induction medium, osteogenic differentiation medium combined with BMP2, osteogenic differentiation combined with inflammation-stimulated TNF (10 ng / mL) , Observe the osteogenic differentiation. MiRNA-specific qRT-PCR was used to observe the changes of miR146a under the above conditions, and then the anti-miR146a antagonist antago-miR146a was transfected into the cells to observe the osteogenic differentiation of mesenchymal stem cells under inflammatory conditions. Results: A stable BMSCs culture system was established. Under different induction conditions, it could form osteogenic, adipogenic and cartilaginous. BMP2 can promote MSC osteogenic differentiation, and mimic inflammation stimulated TNF can inhibit the differentiation of MSCs; differentiated miR146a expression decreased; and TNF dose-dependent enhanced miR146a expression; miR146a antagonist can reverse TNF-induced bone marrow Stem cell osteogenic differentiation decreased. Conclusion: miR146a is involved in the inhibition of osteogenic differentiation of mesenchymal stem cells under inflammatory conditions.