酪氨酸蛋白激酶抑制剂和细胞松弛素D对应力下成骨细胞ERK1/2早期变化的调控  

The regulation of tyrosine protein kinase blocking agent and cytochalasin D to ERK1/2 under the mechanical force

作　　者：曹阳[1] 郑翼[2] 罗颂椒[3] 

机构地区：[1]中山大学光华口腔医学院.附属口腔医院.口腔医学研究所,广州510055 [2]珠海思迈口腔研究中心 [3]四川大学华西口腔医学院正畸科

出　　处：《中华口腔医学研究杂志（电子版）》2008年第4期24-27,共4页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：广东省自然科学基金博士启动项目(5300622)

摘　　要：目的观察机械应力下酪氨酸蛋白激酶(TPK)抑制剂和细胞松弛素D对体外培养的成骨样细胞细胞外信号调节激酶(ERK)1/2早期表达变化的影响。方法采用四点弯曲细胞力学加载装置系统,对人成骨样细胞进行加力:频率0.5Hz,加力板变形量4000 μstrain牵张应力作用成骨样细胞(MG-63)5min时,使细胞发生单轴牵张和压缩应变。运用Western免疫印迹检测不同方向应力应变下TPK抑制剂和细胞松弛素D对ERK1/2表达变化的影响。结果TPK抑制剂预处理后,牵张应力激活ERK的作用被明显阻断,与单纯张力组相比差异有统计学意义(P<0.01);而压缩应力的诱导作用未受影响(P>0.05);低剂量的细胞松弛素D处理后,压应力的诱导作用被明显阻断,ERK的磷酸化水平甚至低于基态,与单纯压力组相比差异有统计学意义(P<0.01),而牵张应力对ERK的激活仅受到一定程度的影响,与单纯张应力组相比差异有统计学意义(P<0.05)。结论力学信号可同时激活细胞内的细胞骨架、TPK转导通道,但各自主要激活的部分存在差别,无论是牵张还是压缩,成骨细胞对力学信号的感知和转导都与微丝细胞骨架密切相关,TPK的活化可能是牵张应力引起黏附斑复合物形成后的主要后续反应,压缩应力的主要后续反应与之关系不大。Objective To study the initial responses and the effect of mechanical strain with tyrosine protein kinase TPK and cytoehalasin D on the expression of ERK1/2 in osteohlast-like cells in vitro. Methods Osteoblast-like cells were cultured with tyrosine protein kinase TPK and cytochalasin D and then subjected to mechanical strain by self-made four-point bending system at a0.5Hz frequency for 5 rain, In each time-phase, the cells were loaded with tension and compression stress at 4000 μstrain respectively. The phosphorylation levels of ERK1/2 were measured by Western Blotting. Changes in the experiments were expressed as ratio to the controls （P 〈 0.05）. Results With tyrosine protein kinase TPK blocking agent 4000 μstrain tensile stimulus could induce the expression of ERK in a few minutes, while compressive stimulus could not. On the contrary, the effect with cytochalasin D of compressive tension strain stimulus was more significant. Conclusions The reception and transduetion of osteoblasts to the mechanical signal of tension or compressive strain are relative to the microfilament cytoskeleton, while the activation of tyrosine protein kinases is mainly connected to response to the tension strain.

关 键 词：机械应力 细胞外信号调节激酶 酪氨酸蛋白激酶 细胞松弛素 

分 类 号：R783.5[医药卫生—口腔医学]