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目的:研究犬骨髓基质细胞(bone marrow stromal cells,BMSCs)在柚皮苷诱导下定向成骨及成骨分化能力。方法:抽取犬骨髓,贴壁法体外培养,流式细胞仪鉴定,加入不同浓度柚皮苷(1×10-5、1×10-6、1×10-7、1×10-8、1×10-9 mol/L)诱导,以CCK-8检测药物毒性,定量检测碱性磷酸酶、von Kossa检测钙结节形成。采用SPSS20.0软件包对数据进行统计学分析。结果:经流式细胞仪检测,CD34、CD45低表达、CD90高表达分别为0.126%、0.075%和95.4%;柚皮苷浓度在10-6、10-7 mol/L时,能明显促进细胞增殖;经柚皮苷诱导后,碱性磷酸酶含量显著提高,其中,浓度为10-6 mol/L时,ALP相对值最高(P<0.05);von Kossa钙结节检测呈阳性。结论:犬骨髓基质细胞在柚皮苷诱导下能分化为成骨细胞。柚皮苷浓度为10-6 mol/L时,能明显促进骨髓基质细胞的增殖及分化。
Objective: To investigate the ability of bone marrow stromal cells (BMSCs) to induce osteogenesis and osteogenic differentiation induced by naringin. Methods: Canine bone marrow was collected and cultured in vitro by adherence method. Flow cytometry was used to determine the concentration of naringin (1 × 10-5, 1 × 10-6, 1 × 10-7, 1 × 10-8, 1, × 10-9 mol / L). The drug toxicity was detected by CCK-8. Alkaline phosphatase was detected quantitatively and von Kossa was used to detect calcium nodule formation. SPSS20.0 software package for statistical analysis of the data. Results: The expression of CD34, CD45 and CD90 were 0.126%, 0.075% and 95.4%, respectively. The concentration of naringin at 10-6 and 10-7 mol / L significantly increased the percentage of cells After alkalin induction, the content of alkaline phosphatase increased significantly. The relative value of ALP was the highest (P <0.05) at the concentration of 10-6 mol / L, while the positive rate of von Kossa calcium nodule was detected. Conclusion: Canine bone marrow stromal cells can differentiate into osteoblasts under the induction of naringin. Naringin concentration of 10-6 mol / L, can significantly promote the proliferation and differentiation of bone marrow stromal cells.