0.2 mT工频磁场引起人羊膜成纤维细胞微丝骨架重组  

Effect of 50 Hz power frequency magnetic field on microfilament cytoskeleton assembly of human amnion FL cells

作　　者：褚克平[1] 蔡知音[1] 张丹英[2] 曾群力[2] 张玉焜[1] 陈树德[1] 夏若虹[1] 

机构地区：[1]华东师范大学物理系 光谱学与波谱学教育部重点实验室,上海200062 [2]浙江大学医学院浙江省生物电磁学重点研究实验室

出　　处：《中华预防医学杂志》2007年第5期391-395,共5页Chinese Journal of Preventive Medicine

基　　金：国家自然科学基金(50137030)

摘　　要：目的探讨50 Hz 工频磁场对人羊膜成纤维细胞微丝装配的影响,并分析磁场对肌动蛋白和表皮生长因子受体蛋白表达的影响。方法将人羊膜成纤维细胞分别用0.1、0.2、0.3、0.4、0.5 mT,50 Hz 工频磁场辐照30 min,以异硫氰四甲基罗丹明-鬼笔环肽标记细胞微丝并在显微镜下观察分析微丝形态的变化。用荧光光谱及紫外光谱检测法测定细胞内微丝总含量,用激光共聚焦显微镜逐层扫描的方法观测细胞微丝骨架的高度,用扫描电镜观测细胞外部形态等方法进一步分析磁场对细胞微丝骨架及细胞形态的影响。用 Western blotting 检测去垢剂不可溶的蛋白的表达量以分析磁场作用的可能机制。结果人羊膜成纤维细胞经不同强度工频磁场辐照30 min 后,只有0.2 mT 工频磁场辐照显著导致细胞中平行排列的微丝应力纤维减少,诱导细胞周边出现致密微丝外周带并伸出丝状伪足,停止辐照2 h 后微丝形态恢复,伪足消失。光谱检测发现胞内微丝总含量无显著变化。激光共聚焦显微镜逐层扫描观测到0.2 mT 磁场辐照使细胞微丝骨架平均高度由(12.37±1.28)μm降低到(9.97±0.38)μm(t=6.96,P>0.05)。扫描电镜图像显示磁场辐照使细胞更为扁平,有足状伪足出现。Western blotting 检测到磁场辐照后细胞骨架中去垢剂不可溶部分的肌动蛋白和表皮生长因子受体含量与对照组相比,分别升高(16.8±2.3)%(t=16.68,P<0.05)和(31.2±4.1)%(t=17.10,P<0.05)。结论 0.2 mT 工频磁场短时辐照影响了人羊膜成纤维细胞微丝骨架的形态,此效应可随磁场撤销而解除,且可能与磁场诱导了细胞膜上表皮生长因子受体聚簇有关。Objective Investigations were carried out to understand the effect of 50 Hz power frequency magnetic field on microfilament assembly of human amniotic cells and on expression of actin and epidermal growth factor receptor. Methods Human amnion FL cells were exposed to 0. 1,0. 2, 0. 3, 0. 4, 0. 5 mT power frequency magnetic field for 30 minutes. Microfilaments were marked using Phalloidin- TRITC, and then were observed under a fluorescence microscope. An optical method was used to detect the relative content of microfilament in cells. A scanning electron microscope was used to detect the cell shape. The content of actin and epidermal growth factor receptor in the preparation of the detergent-insoluble cytoskeleton were measured by western-blotting to analyse the potential mechanism of the change induced by magnetic field. Results Intracellular stress fibers were found to decrease after exposing cells to a 0. 2 mT power frequency magnetic field for 30 minutes. New microfilament and filopedia bundles appeared at the cell periphery after exposure, but the detected total F-actin content per cell was not significantly changed, detected by a F-actin-specific dye. The change in the amount of microfilaments caused by the field could be recovered 2 hours later when the field was withdrawn. The mean height of microfilament cytoskeleton decreased from （12.37±1.28） μm to （9.97±0.38） μm （t =6.96, P 〉0.05） after exposure using a confocal microscope. The cell shapes became more flat and lamellipodia appeared after exposure observed by a scanning electron microscope. By using Western blotting method, the intracellular contents of epidermal growth factor receptor and of actin in the preparation of the detergent-insoluble cytoskeleton which are associated with high-affinity epidermal growth factor receptors, increased about （31.2±4. 1 ）% （t = 17.10, P 〈 0. 05 ） and （ 16. 8 ± 2. 3 ） % （ t = 16. 68, P 〈 0.05 ） respectively, compared with that of the control. Conclusion These results suggest that

关 键 词：电磁场 微丝 受体 表皮生长因子 张力纤维 

分 类 号：R730.2[医药卫生—肿瘤]