朱波，吴华，黄珊珊，赵东明.电磁场作用下大鼠骨髓间充质干细胞的成骨分化与增殖[J].中华物理医学与康复杂志,2015,(10):727-732
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| 电磁场作用下大鼠骨髓间充质干细胞的成骨分化与增殖 |
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| DOI： |
| 中文关键词: 电磁场 间质干细胞 丝裂原激活蛋白激酶类 p38丝裂原活化蛋白激酶类 |
| 英文关键词: Electromagnetic fields Mesenchymal stem cells Mitogen-activated protein kinases p38 Mitogen-activated protein kinases |
| 基金项目:国家自然科学青年基金项目(81301083)；基金面上项目(51077065) |
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| 中文摘要: |
| 目的研究15Hz 1mT的正弦波电磁场对大鼠骨髓间充质干细胞（MSCs）的细胞外信号调节激酶（ERK）和丝裂原激活的蛋白激酶（MAPK）p38的激活规律及相互作用特点，探讨ERK和p38 MAPK信号通路在电磁场促成骨效应中的作用。 方法选取第3代体外分离培养的大鼠骨髓间充质干细胞，分为对照组、曝磁组、曝磁+PD98059组和曝磁+SB202190组四个大组。曝磁组细胞置于带有电磁发生器的培养箱中培养，曝磁+PD98059组和曝磁+SB202190组细胞分别加入ERK信号通路阻断剂PD98059和p38 MAPK信号通路阻断剂SB202190后再置入带有电磁发生器的培养箱中培养，对照组细胞正常培养。用免疫印迹（Western blot）法检测ERK和p38 MAPK的蛋白表达及磷酸化水平变化。按照细胞碱性磷酸酶（ALP）试剂盒说明书操作步骤对各组细胞ALP活性进行检测，其活性变化可间接反映细胞的分化成骨活性；用噻唑蓝比色法检测各组细胞的增殖活性变化。 结果①电磁场作用下，骨髓间充质干细胞内p38 MAPK信号通路可被快速激活，曝磁15min后出现p38磷酸化水平升高（P<0.05）；加用SB202190后，再行电磁场刺激，细胞内p38磷酸化水平仍然维持在较低水平，与曝磁组比较，差异有统计学意义（P<0.05）。②与对照组比较，曝磁5d后细胞内ALP活性显著升高(P<0.05)，SB202190可明显阻断该效应(P<0.05)。③与对照组比较，曝磁3d后骨髓间充质干细胞的增殖活性明显升高(P<0.05)，SB202190不能阻断该效应(曝磁+SB202190组与曝磁组比较，P>0.05)。④SB202190阻断p38 MAPK信号通路并曝磁5min后，ERK MAPK磷酸化水平明显强化(P<0.05)；PD98059阻断ERK MAPK信号通路并曝磁30min后，p38 MAPK磷酸化水平明显强化(P<0.05)。 结论ERK和p38 MAPK信号通路分别参与了电磁场对骨髓间充质干细胞增殖和分化成骨过程的调节，并在电磁场作用下两通路表现出串扰现象。 |
| 英文摘要: |
| Objective To study the effects of electromagnetic field (EMF) on the activation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) pathways in mesenchymal stem cells (MSCs) and their interaction, and to explore the cellular signal transduction mechanism of the biological effects induced by EMF. MethodsThe 3rd-passage rat bone marrow MSCs were randomly divided into a control group, an EMF group, an EMF+PD98059 group and an EMF+SB202190 group. Cells in the EMF group were cultured in the electromagnetic field, those in the EMF+PD98059 and EMF+SB202190 groups cultured in the electromagnetic field after PD98059 or SB202190 was added, and those in the control group were cultured normally. Different groups of cells were exposed to electromagnetic fields (15 Hz, 1 mT, sine wave form) for different exposure duration. The activated phosphorylated and non-phosphorylated p38MAPK were measured using Western blotting analysis with their specific corresponding antibodies. The alkaline phosphatase (ALP) activity in cells in different groups was detected according to the instructions of ALP kit. MTT assay was applied to investigate the proliferation of cells. ResultsElectromagnetic fields could rapidly induce the activation of p38 MAPK (P<0.05) and the phosphorylation of p38 MAPK elevated after 15 min exposure to EMF. The phosphorylation of p38 MAPK was significantly lower in the EMF+SB202190 group than that of the EMF group. After 5 days of EMF exposure, the ALP activity of cells was significantly improved, and the effect could be inhibited by SB202190. The bone marrow mesenchymal stem cells proliferation increased significantly after being exposed to EMF for 3 days, and it could not be blocked by SB202190. Phosphorylation of ERK and MAPK increased significantly when the p38 MAPK pathway was blocked by SB202190 and exposed to EMF for 5 minutes, and it also increased significantly when the ERK MAPK pathway was blocked by PD98059 and received 30 minutes of EMF exposure. ConclusionEMF can quickly activate ERK and p38 MAPK pathways to induce cell proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. Moreover, in EMF there is a mutual interference between ERK and p38 MAPK pathways. |
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