摘要
目的:观察外源性电场对培养人视网膜色素上皮humanretinalpigmentepithelial,hRPE)细胞移行方向及细胞内肌动蛋白表达的影响。方法:培养于DMEM+100g/LFBS的hRPE细胞暴露于强度为6V/cm的电场中,未暴露于电场的细胞作为对照。观察并记录2h中每隔15min细胞移行的变化图像,测量细胞移行距离、细胞移行方向及其与场线间夹角。2h后采用间接免疫荧光法检测实验组与对照组细胞内肌动蛋白表达情况。SPSS10.0及Image-ProPlus5.0软件处理结果。结果:未暴露于电场中的hRPE细胞在观察期间呈无序移行,2h后细胞分布未见特殊变化,细胞内肌动蛋白有弱阳性表达。细胞暴露于电场30min后开始出现阴极方向的移行趋势,大部分细胞长轴趋向垂直于场线方向排列。细胞的移行表现为移行速度和移行方向的一致性均随时间延长而增加(P<0.05)。暴露于电场2h后,大部分细胞内肌动蛋白表达阳性,且多集中分布于细胞质内朝向电场阴极一侧。结论:外加电场可诱导hRPE细胞向电场阴极方向定向移行,同时伴有细胞内肌动蛋白阳性表达及定向分布。电场对细胞的作用与暴露时间成正相关。视网膜脱离后的内源性电场的存在可能参与并诱导了RPE细胞的定向移行。
AIM: To investigate the effect of adscititious electric field on the migratio n direction and actin expression of cultured human retinal pigment epithelial (h RPE) cells. · METHODS: hRPE cells cultured in DMEM+100g/L FBS were exposed to the electric field at 6V/cm, and those unexposed to electric fields were used as controls. Se rial images of the cells were taken at 15min intervals within 2h, and the displa cement of the cells and the angle between the cellular translocation direction a nd the field vector were measured. After 2h, indirect immunofluorescence method was applied to detect the expression of actin in the cells. The results were ana lyzed by SPSS 10.0 and Image-Pro Plus5.0 software. · RESULTS: hRPE cells unexposed to electric field moved randomly and showed no special distribution after 2h, and there was weak expression of actin in the cel ls. After 30min in the electric field, most of the cells oriented their long axe s perpendicular to the electric field and began to migrate to the cathode. The d irected migration of cells was manifested by the fact that the migration velocit y and the coherence of the migration direction increased with the process of exp osure time (P < 0.05). After exposed to electric field for 2h, the expression of actin became positive in most of the cells, with the distribution of the protei n to the cathodal side. · CONCLUSION: Electric field could induce directed migration of hRPE cells towa rds cathode, with positive expression and directed distribution of actin. This e ffect was positively correlated with the exposure time of the cells in the elect ric field. Endogenous electric field may participate in and induce the directed migration of RPE cells after the retinal detachment. ·
出处
《国际眼科杂志》
CAS
2004年第6期998-1001,共4页
International Eye Science
基金
中国国家自然科学基金项目(No.30200311)
关键词
人视网膜色素上皮细胞
电场
定向移行
实验研究
electric field
retinal pigment epithelium
cell migration
actin
cytoskeleton
