三磷酸腺苷促进骨髓间充质干细胞成骨分化的机制  

Osteogenic differentiation of bone marrow-derived mesenchymal stem cell promoted by adenosine triphosphat

作　　者：杨勇[1] 李文凯[1] 宋明宇[1] 吴华[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院骨科,武汉430030

出　　处：《中华骨质疏松和骨矿盐疾病杂志》2015年第3期236-242,共7页Chinese Journal Of Osteoporosis And Bone Mineral Research

基　　金：国家自然科学基金青年基金项目(81301552)

摘　　要：目的探讨三磷酸腺苷(adenosine triphosphate,ATP)对骨髓间充质干细胞增生和向成骨分化的影响及初步机制。方法体外培养SD大鼠骨髓间充质干细胞,取第3代细胞,分对照组和ATP作用组,加入一定浓度的ATP(600、300、150μmol)。用半定量反转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)和real time PCR法检测多个成骨指标mRNA的表达,对硝基苯磷酸盐(ρ-nitrophenylphosphate,PNPP)法检测碱性磷酸酶的表达,Von kossa法检测钙结节的形成。结果 ATP组碱性磷酸酶活性升高(P<0.05),多个成骨指标如BMP2、BSP、DLX等表达升高(P<0.05),ATP组钙结节数量明显多于对照组。在加入P2X7受体抑制剂后,ATP促成骨指标BMP2、BSP、DLX等的表达有所减弱(P<0.05)。结论一定浓度的ATP促进骨髓间充质干细胞碱性磷酸酶的表达、多个成骨指标的表达、钙结节的形成,以及促进骨髓间充质干细胞向成骨分化,并受P2X7受体的影响。Objective To study the mechanism and the osteogenesis differentiation of bone marrow mesenchymal stem cells stimulated by adenosine triphosphate（ ATP）. Methods The bone marrow mesenchymal stem cells of SD rats were isolated and cultured in vitro. The third passage cells were harvested and added with ATP. The semi-quantitative reverse transcription-polymerase chain reaction（ RT-PCR） and real time PCR were used to measure the osteogenic marker expression; The ρ-nitrophenylphosphate（ PNPP） method was used to measure alkaline phosphatase（ ALP） activity.The calcium nodes were measured by Von kossa stain. Results ATP induced a significant increase in expression of ALP,RUNX2,bone morphogenetic protein 2（ BMP2）（ P〈0. 05）,and calcium node formation in comparison to the controls.The osteogenesis marker expression was reduced by P2X7 inhibitor brilliant blue G（ BBG）. Conclusion ATP promoted osteogenesis differentiation of bone marrow mesenchymal stem cells. The effect was correlated P2X7 receptor.

关 键 词：三磷酸腺苷 骨髓间充质干细胞 成骨分化 

分 类 号：R681[医药卫生—骨科学]