电磁场激活细胞外信号调节激酶通路在骨髓间充质干细胞增殖与分化成骨中的作用  

The effects of activating the ERK signaling pathway with an electromagnetic field on osteogenic proliferation and differentiation of mesenchymal stem cells

作　　者：赵东明[1] 吴华[1] 黄珊珊[2] 黄飞[1] 赵文春[3] 杨勇[1] 陶超雄[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 [2]华中科技大学同济医学院组织胚胎学教研室 [3]海军工程大学电气与信息工程学院

出　　处：《中华物理医学与康复杂志》2008年第8期515-518,共4页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：国家自然科学基金面上项目（50477043）

摘　　要：目的探讨电磁场作用于大鼠骨髓间充质干细胞对细胞外信号调节激酶（ERK）信号通路的影响，并进一步研究电磁场诱导所致ERK信号通路变化在骨髓间充质干细胞增殖与分化成骨中的作用。方法体外培养大鼠骨髓间充质干细胞，取第3代细胞，分为对照组、暴磁组、PD98059组和PD98059＋暴磁组。采用Westernblotting法检测ERK通路的活性变化，采用四甲基偶氮唑盐（MTT）法检测各组细胞的增殖活性，按照碱性磷酸酶（ALP）试剂盒说明书步骤检测各组细胞ALP活性。结果①暴磁后5min，ERK1／2磷酸化水平即明显高于对照组，1h后仍处于较高水平（P〈0．01）；PD98059作用可明显抑制ERK1／2磷酸化水平的升高，提示PD98059可有效阻断ERK通路的激活。②MTT法检测结果提示，暴磁后细胞的增殖活性明显升高，PD98059可明显抑制这种效应。③暴磁组ALP活性明显高于对照组，PD98059＋暴磁组ALP活性较暴磁组高（P〈0．01），差异有统计学意义。结论在15Hz、1mT的正弦波电磁场作用下，骨髓间充质干细胞ERK信号通路很快被激活，引起细胞增殖活性和ALP活性的改变，这为进一步研究磁场对骨髓问充质干细胞的促增殖和分化成骨机制提供了指导意义。Objective To investigate the effects of an electromagnetic field on the extra-cellularly regulated kinase （ERK） signalling pathway and to determine the impact of electromagnetic activation on osteogenie proliferation and differentiation in rat bone marrow mesenchymal stem cells. Methods Rat bone marrow mesenchymal stem cells were isolated and cultured in vitro. The third-passage cells were divided into 4 groups （ Control, PD98059, EMF and EMF ＋ PD98059）. Western blotting was used to detect the activation of the ERK signal pathway after exposure to an electromagnetic field. MTT assay was used to determine the activation of proliferation in the cells in the different groups. The cells＇ alkaline phosphatase activities were also detected. Results （1） The ERK signal pathway in these rat bone marrow mesenchymal stem cells was activated after exposure to a 15 Hz, 1 roT, sine wave form electro- magnetic field for 5 rain. Activation remained high for at least 1 h. PD98059 can effectively block the activation of the ERK signal pathway. （2） Cell proliferation was promoted after exposure to the electromagnetic field, and this effect could be significantly inhibited by PD98059. （3） Alkaline phosphatase was significantly elevated in these bone mar- row mesenchymal stem cells after exposure to the electromagnetic field. The activation in the EMF ＋ PD98059 group was slightly greater than in the EMF group. Conclusion Electromagnetic fields of 15 Hz and 1 mT can activate the ERK signal pathway and alter proliferation and osteogenic differentiation in the bone marrow mesenchymal stem cells of rats.

关 键 词：电磁场 细胞外信号调节激酶信号通路 骨髓间充质干细胞 细胞增殖 分化成骨 

分 类 号：R68[医药卫生—骨科学]