次声暴露对血管内皮细胞骨架微丝F-actin表达的影响

Effects of exposure to infrasound on the expression of cytoskeleton filament F-actin in vascular endothelial cell-304

目的研究16Hz,90,110及130dB的次声作用对人脐血管内皮细胞(ECV-304)骨架F-actin表达的影响。方法将ECV-304接种于细胞爬片上,并将其分为对照组和90,110,130dB次声暴露组。各次声暴露组均接受2h相应强度的次声作用,对照组则作次声假暴露处理。于次声作用后即刻、1h、2h、4h、8h、12h及24h分别对各组细胞进行F-actin免疫荧光染色检测,同时应用激光扫描共聚焦显微镜观察各组细胞F-actin的表达变化,记录并测定其F-actin的平均荧光强度。结果对照组细胞中的大部分荧光样物质呈弥漫状态,胞膜荧光较强,胞浆内可见少量肌动蛋白纤维丝,方向不规则,长短不一。即刻观察经不同声压级次声作用后的3组细胞,均可见其胞浆中微丝F-actin明显粗大、变长,其间的荧光样物质大多为较长的粗大应力丝,沿细胞纵轴排列较多,细胞数量及荧光强度明显增加,细胞膜与对照组类似,均结构完整且荧光增强;在次声作用后8h时,各次声暴露组细胞的F-actin仍处于高表达状态;随着时间的延长,其F-actin表达逐渐降低;当次声作用后24h时,各次声暴露组F-actin与对照组已无显著性差异。不同声压级次声暴露组细胞的F-actin变化趋势均基本一致,3组细胞的F-actin表达在各检测时间点均未见显著性差异。结论16Hz,90dB、110dB及130dB的短时次声暴露均可诱导人脐血管内皮细胞F-actin表达改变,导致其骨架重建;并且由短时次声作用诱发的细胞骨架改建可在次声暴露结束后24h时恢复正常。

Objective To investigate the effects of exposure to infrasound of different intensities on the expression of F-actin in vascular endothelial cell-304 （ECV-304）. Methods The ECV-304 primary culture cells were exposed to intTasound of 0 dB （the control group） , 90 dB, 110 db, 130 dB, respectively, for 2 hours. Laser scanning confocal microscope was used to examine the changes of F-aetin after immunofluorescent staining at 0 h, 1 h, 2 h, 4 h, 8 h, 12 h and 24 h after treatment and the photos were taken for further.analysis of the cell average fluorescence. Resuits In the control grnup, most fluorescein-labelled substance was scattered in the cells, and few actin filaments were observed with irregular alignment, while the expression of F-actin in the experimental groups was thicker, longer and showed in longitudinal arrangement after exposure, and the intensity of flunrescence was significantly increased, In all groups, complete and strong fluorescence was observed at cellular membrane. The over-expression of F-actin in the experimental groups was kept obviously for 8h, however there was no significant difference of the F-actin expression between the experimental groups and the control group 24 h later. The expression of F-actin in all the experimental groups was synchronous, and there was no obvious difference at each time point, Conclusion F-actin expression in ECV- 304, exposed to infrasound with intensity output of 90 dB, 110 dB, 130 dB and frequency of 16 Hz, cnuld be changed, and recuperated 24 h after infrasound exposure.