骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞成骨分化潜能的影响

拟研究骨碎补总黄酮对缺氧环境中犬骨髓间充质干细胞(BMSCs)成骨分化潜能的影响.运用骨碎补总黄酮(TFDR)干预低氧浓度(10％)环境中犬BMSCs 4周后诱导成骨分化,倒置显微镜观察茜素红染色BMSCs钙结节形成,比色法检测碱性磷酸酶(ALP)活性水平,流式细胞术检测细胞线粒体膜电位,激光共聚焦显微镜和RT-PCR...

Effects of Total Flavonoids of Drynariae Rhizoma on Osteogenic Differentiation Potential of Canine Bone Marrow Mesenchymal Stem Cells in Hypoxic Environment

This study aims at studying the effects of the total flavonoids of Drynaria Rhizoma (TFDR) on osteogenic differentiation potential of canine bone marrow mesenchymal stem cells (BMSCs) in hypoxic. TFDR was used to intervene canine BMSCs in a low oxygen concentration (10%) environment for 4 weeks to induce osteogenic differentiation. Observation of calcium nodule formation in BMSCs stained with Alizarin Red by an inverted microscope. The alkaline phosphatase (ALP) activity level was detected by colorimetric method. The cell mitochondrial membrane potential was detected by flow cytometry. The expression of Runx2 and Osterix, the key genes for osteogenic differentiation were detected by RT-PCR and confocal laser microscope. Results were as follows: Compared with the control group, 10% oxygen concentration inhibited the formation of calcium nodules and ALP activity in canine BMSCs (P<0.01), Mitochondrial membrane potential level dropped (P<0.05), and the expression of Runx2 and Osterix decreased (P<0.05 or P<0.01). Compared with the hypoxic group, TFDR can significantly promoted the formation of calcium nodules and ALP activity of BMSCs in Hypoxic (P<0.05), the level of cell mitochondrial membrane potential increased (P<0.05), and Runx2 and Osterix expression increased (P<0.05 or P<0.01). TFDR can promote the osteogenic differentiation of canine BMSCs in hypoxic.