EphB6在小鼠骨髓间充质干细胞成骨分化中的初步研究  

Effect of EphB6 on osteogenic differentiation of bone marrow-derived mesenchymal stem cells

作　　者：李松涛[1] 刘涌[1] 宋磊[1] 董世武[2] 邸宁[1] 李殿威[1] 徐源[1] 周强[1] 

机构地区：[1]第三军医大学西南医院骨科,全军矫形外科中心,重庆400038 [2]第三军医大学基础医学部人体解剖学教研室

出　　处：《第三军医大学学报》2012年第10期907-911,共5页Journal of Third Military Medical University

基　　金：国家高技术研究发展计划(863计划)(2006AA02Z4E3);国家自然科学基金(81027005)~~

摘　　要：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 effect of EphB6 on osteogenic differentiation of mouse bone marrow-derived mesenchymal stem cells（BMSCs）.Methods The BMSCs seeded on decalcified bone matrix by biphasic seeding were divided into an osteogenic differentiation 5-day group（group A）,an osteogenic differentiation 14-day group（group B）,a non-induced 5-day group（group C） and a non-induced 14-day group（group D）.Alizarin red staining was performed to detect calcium nodules.The activity of alkaline phosphatase（ALP） and mRNA expression of Runx2,Osterix and EphB6 were determined by ALP kit and quantitative real-time PCR,respectively.The indicators above were examined again after the BMSCs were treated with soluble ephrinB1-Fc to activate EphB6 in all the groups.Results ALP activity and mRNA expression of Runx2 and Osterix increased in group A and group B,and ALP activity（7.32±2.02 King＇s unit/100 ml） and mRNA expression of Runx2（3.442 6±0.258 5） and Osterix（2.482 3±0.329 2） reached the peak levels in group B.EphB6 mRNA expression was downregulated in group A and group B,and reached the bottom level in group B（0.754 3±0.023 5）.After the treatment of soluble ephrinB1-Fc for EphB6 activation,a significant reduction of calcium nodules was observed and ALP activity significantly decreased（P0.05）.High-dose ephrinB1-Fc showed stronger ALP activity decrease effects than low-dose ephrinB1-Fc（12.20±1.30 vs 15.40±1.03 King＇s unit/100 ml,P=0.008）.Similarly,the mRNA expression of Runx2 and Osterix was downregulated under ephrinB1-Fc treatment in a dose-dependent manner.Conclusion EphB6 inhibits the osteogenic differentiation of BMSCs via mRNA expression regulation of Runx2 and Osterix.

关 键 词：EPHRIN RECEPTOR 骨髓间充质干细胞 成骨分化 组织工程 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学] R341[医药卫生—基础医学]