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背景:前期研究发现,黄精多糖通过Wnt/β-catenin信号通路促进骨髓间充质干细胞成骨分化,但是其促进骨髓间充质干细胞成骨分化的作用及分子机制尚不明确。目的:探讨黄精多糖LRP5激活Wnt信号通路促进小鼠骨髓间充质干细胞向成骨细胞分化的研究。方法:培养C57BL/6小鼠骨髓骨髓间充干细胞,构建LRP5干扰载体,转染C57BL/6小鼠骨髓间充质干细胞,转染后在荧光倒置显微镜下计算细胞荧光转染效率及Western Blot检测LRP5蛋白的表达。碱性磷酸酶、茜素红染色和Western Blot分析转染LRP5-siR NA病毒后小鼠骨髓间充质干细胞成骨能力的改变;RT-PCR及双荧光素酶检测黄精多糖对沉默LRP5的小鼠骨髓间充干细胞的成骨分化作用。结果与结论:(1)与对照组相比,LRP5转染组细胞显著降低细胞的矿化能力(P<0.05),明显降低Osterix、Runx2基因及LRP5蛋白的表达(P<0.05);(2)黄精多糖能促进转染LRP5-siR NA病毒的小鼠骨髓间充质干细胞向成骨细胞分化,能显著上调β-catenin及Osterix基因的表达(P<0.05),促使含有TCF结合位点的荧光素酶报告基因(TOPFlash)的高表达(P<0.05);(3)结果说明,LRP5在小鼠骨髓间充质干细胞的成骨分化过程中有重要作用。黄精多糖不依赖于LRP5激活Wnt/β-catenin信号通路促进小鼠骨髓间充质干细胞向成骨细胞分化。
BACKGROUND: Preliminary studies have found that Polygonatum Polysaccharides promote the osteogenic differentiation of BMSCs through the Wnt / β-catenin signaling pathway, but the role and molecular mechanism of Polygonatum Polysaccharide in promoting the osteogenic differentiation of BMSCs remains unclear. OBJECTIVE: To investigate the effect of Polygonatum polysaccharide LRP5 on Wnt signaling pathway to promote the differentiation of mouse bone marrow mesenchymal stem cells into osteoblasts. Methods: C57BL / 6 mouse bone marrow marrow stromal cells were cultured, LRP5 interference vector was constructed and transfected into C57BL / 6 mouse bone marrow mesenchymal stem cells. The transfection efficiency was calculated by inverted fluorescence microscope and the transfection efficiency was calculated by Western Blot LRP5 protein expression. Alkaline phosphatase, alizarin red staining and Western Blot were used to analyze the osteogenic capacity of mouse bone marrow mesenchymal stem cells after transfection with LRP5-siR NA virus. RT-PCR and dual luciferase assay showed that polygalacturonase Osteogenic differentiation of murine bone marrow stromal cells. RESULTS AND CONCLUSION: (1) Compared with the control group, LRP5 transfected cells significantly decreased the mineralization ability of cells (P <0.05) and significantly decreased the expression of Osterix, Runx2 and LRP5 protein (P <0.05); (2 ) Polygonatum polysaccharides could promote the differentiation of mouse bone marrow mesenchymal stem cells transfected with LRP5-siR NA into osteoblasts, significantly up-regulate the expression of β-catenin and Osterix genes (P <0.05) (P <0.05). (3) The results showed that LRP5 plays an important role in the osteogenic differentiation of mouse bone marrow mesenchymal stem cells. Polygonatum Polysaccharides Promote Differentiation of Mouse Bone Marrow Mesenchymal Stem Cells into Osteoblasts Independent of LRP5-activated Wnt / β-catenin Signaling Pathway.