论文部分内容阅读
目的探讨EphB6在小鼠骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMSC)成骨分化中的作用及其可能机制。方法将BMSC通过双相接种法接种于脱钙骨基质,分为4组:成骨诱导5、14 d(A、B组),普通培养5、14 d(C、D组),茜素红染色检测钙结节,定量检测各组碱性磷酸酶活性,Runx2、Osterix和EphB6的mRNA表达,采用游离态配体ephrinB1-Fc激活EphB6并重复检测上述指标。结果 ALP活性、Runx2和Osterix在A、B组增高,且B组检测到最高的ALP活性[(7.32±2.02)金氏单位/100 ml]、Runx2(3.442 6±0.258 5)倍和Osterix(2.482 3±0.329 2)倍。EphB6的表达水平在A、B组降低,且在B组最低(0.754 3±0.023 5)倍。以ephrinB1-Fc激活EphB6后,茜素红染色显示钙结节明显减少,ALP活性显著下降(P<0.05),在高浓度配体作用后ALP活性[(12.20±1.30)金氏单位/100 ml]较低浓度配体[(15.40±1.03)金氏单位/100 ml]有进一步下降(P=0.008)。配体作用后,BMSC成骨分化中Runx2和Osterix的表达显著下降。结论 EphB6通过调控Runx2和Osterix抑制BMSC成骨分化。
Objective To investigate the role of EphB6 in osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSC) and its possible mechanism. Methods BMSCs were inoculated into decalcified bone matrix by biphasic inoculation and divided into 4 groups: osteoblasts for 5 and 14 days (groups A and B), normal culture for 5 and 14 days (groups C and D), alizarin red The calcium nodules were detected by staining. The alkaline phosphatase activity, mRNA expression of Runx2, Osterix and EphB6 in each group were detected quantitatively. EphB6 was activated by free ligand ephrinB1-Fc and the above indexes were detected repeatedly. Results ALP activity, Runx2 and Osterix increased in groups A and B, and the highest ALP activity [(7.32 ± 2.02) units / 100 ml], Runx2 (3.442 6 ± 0.258 5) and Osterix (2.482 3 ± 0.329 2) times. The expression of EphB6 decreased in groups A and B, and was lowest in group B (0.754 3 ± 0.023 5) times. After activated by ephrinB1-Fc, alizarin red staining showed a significant decrease of calcium nodules and a marked decrease of ALP activity (P <0.05). ALP activity was significantly increased after high concentration of ligand [(12.20 ± 1.30) ] With a lower concentration of ligand [(15.40 ± 1.03) gold units / 100 ml] there is a further decline (P = 0.008). After ligand, the expression of Runx2 and Osterix in osteogenic differentiation of BMSC significantly decreased. Conclusion EphB6 inhibits osteogenic differentiation of BMSCs by regulating Runx2 and Osterix.