不同声强超声波对细胞骨架微丝F-actin表达的影响  

The expressions of cytoskeleton filament F-actin of EVC-304 cells exposed to ultrasound in different intensities

作　　者：王冰水[1] 袁华[1] 王斌[1] 牟翔[1] 刘卫[1] 刘静[1] 陈丹[2] 

机构地区：[1]第四军医大学西京医院物理医学与康复科,西安710032 [2]第四军医大学电镜中心

出　　处：《中国康复医学杂志》2009年第6期515-517,I0001,共4页Chinese Journal of Rehabilitation Medicine

摘　　要：目的:研究不同声强超声波对人脐血管内皮细胞(ECV-304)细胞骨架F-actin表达的影响,探讨超声波对人体的作用机制。方法:将人脐血管内皮细胞ECV-304接种于6孔板中,分别以频率800kHz,占空比为30%功率密度0.20、0.40、0.60和0.80W/cm2,的脉冲超声波辐射5min,于辐射后不同时间,对ECV-304细胞进行F-actin免疫荧光染色,应用激光扫描共聚焦显微镜,观察细胞F-actin表达的改变,测定单个细胞F-actin的平均荧光强度。结果:假辐射组细胞有中等量的F-actin表达,荧光物质部分呈弥漫状态,部分呈线形应力丝状态,细胞膜荧光较强,胞浆内的肌动蛋白纤维丝,方向不规则,长短不一。四组不同强度超声波辐射后的细胞变化较为一致,辐射后均可看到胞浆中微丝F-actin粗大纤长,弥漫状态的荧光物质相对较少,应力丝沿细胞纵轴排列较多,数量及荧光强度也明显增加,细胞膜荧光较强,超声波辐射的四组细胞之间没有明显的差别。随时间的延长细胞应力丝逐渐减少,12h时已接近正常组细胞,各组细胞的变化是同步的。结论:800kHz的脉冲式超声波辐射ECV-304细胞可诱导F-actin表达的改变,导致细胞骨架重建,超声波强度≤0.80W/cm2时,引起的细胞骨架改变是可逆的。Objective：To observe the expressions of F-actin in ECV-304 （human umbilical vein endothelial cells） cells exposed to ultrasound in different intensity outputs. Method：After primary euhure on eoverglass , ECV-304 cells were divided into five groups. In experimental groups, the cells were exposed by ultrasound,frequency of 800kHz,30% duty cycle ,with intensity output of 0.20, 0.40, 0.60 and 0.80W/cm^2 respectively for 5 min. The cells in control group were managed in the same environment as experimental groups but without ultrasound outputs. Laser scanning confocal microscope was used to examine the changes of cytoskeleton filament F-actin after immunofluorescent staining and the photos were taken for further analysis of ceils average fluorescence by spectrofluorimetric quantification. Result：Cells in control group, some fiuorescein-labelled substance was in diffusion states. Less aetin filaments were tenuous, short and with irregular arrangoment. Correspondingly, F-aetin in the cells of experimental groups was thick,long and with longitudinal arrangement after exposure. The intensity of fluorescence increased significantly. There was a membrane-like boundary strong fluorescence. More F-actin expressions in experimental groups were being kept obviously for hours. However, 12h later there was no signifieant difference in F-aetin expressions between experimental groups and control group. Anyway, cells in experimental groups showed synchronous changes. Conclusion：F-actin expressions in ECV-304 cells can be changed by 800kHz ultrasound exposures with different intensity outputs in the experiment. The reversible expressions of F-actin indicates that the changes of cell skeleton reconstruction induced by 0.80W/cm^2 or less ultrasound exposure are reversible.

关 键 词：超声 人脐血管内皮细胞 聚合态肌动蛋白 激光扫描共聚焦显微镜 

分 类 号：R454.3[医药卫生—治疗学]