羟基红花黄色素A对人骨髓间充质干细胞成骨分化的影响

Effects of Hydroxyl Safflor Yellow A on osteogenic differentiation of human bone marrow stromal stem cells

目的:探讨羟基红花黄色素A(HSYA)对骨髓间充质干细胞(BMSCs)体外多潜能性及成骨分化能力的影响。方法:组织贴壁培养法分离培养女性下颌骨来源BMSCs,CCK-8法检测不同浓度的HSYA对BMSCs生长的影响;Western blot及RTPCR方法检测加入HYSA的BMSCs的多潜能转录因子NANOG、SOX2、OCT4的表达,比较两组细胞的克隆形成率;茜素红染色、Western blot、实时定量RT-PCR检测空白组、阳性对照组及实验组的体外成骨分化能力。结果:与对照组相比,实验组多潜能因子NANOG、SOX2及OCT4蛋白及mRNA表达水平升高,差异具有统计学意义;体外成骨诱导7 d后,实验组BMSCs中Runx2等成骨分化基因表达上升,诱导14 d后,钙结节形成较空白组及标准组多。结论:HYSA能促进BMSCs的增殖,同时在Runx2、OCN、OSX、OPN的参与下,诱导其向成骨方向分化。

Objective To investigate the effect of the hydroxyl safflor yellow A in regulating the stemness and the activity of osteo-genic differentiation of bone marrow stromal cells（BMSCs） in vitro. Methods： Tissue adherent culture was used to isolate the BMSCs from female mandibular,and CCK-8 method was used to detect the effect of different concentrations of HSYA on the growth of BMSCs. The stemness transcription factors （NANOG, SOX2,OCT4） were detected by Western blot and RT-PCR in BMSCs supplemented with HYSA. The colony-fomiing efficiency was compared between HYSA group and control group. The different osteogenic ability of the HY- SA group,positive group and the control group was analyzed by real-time RT-PCR,Western blot and Alizarin red staining after osteo-genic induction. Results： Compared with the control group,the expression levels of NANOG,SOX2, and OCT4 of HYSA group were higher than that of the control group, and there was statistically significant difference. Seven days later osteogenic induction in vitro the expression of osteogenic differentiation genes were increased in the experimental group BMSCs. 14 days later osteogenic induction, the calcium nodules of HYSA group were more than that of the control and positive group. Conclusions ： HYSA can promote the prolifera-tive capacity of BMSCs,and induce their osteogenic differentiation,due to the participation of Runx2, OCN, OSX, OPN.