电场对人视网膜色素上皮细胞生物学活性的影响

目的:观察电场作用对培养人视网膜色素上皮(human retinal pigment epithelial,hRPE)细胞活力及分裂过程的影响。方法:hRPE细胞置于强度为8V/cm的直流电场中暴露3h,停止电场作用后继续培养12h;未受电场作用的hRPE细胞作为对照组。显微摄像系统记录各时间点细胞图像,观察细胞形态变化;细胞行台盼蓝拒染活细胞计数及核仁嗜银蛋白(AgNORs)染色,图像分析染色结果;流式细胞仪检测细胞凋亡情况。结果:暴露于电场中的hRPE细胞伸长,垂直于场线方向排列,停止暴露后细胞恢复正常形态及分布;各时间点实验组与对照组细胞的台盼蓝拒染率差异无统计学意义(P>0.05);电场暴露前、暴露3h及停止暴露并继续培养12h后hRPE细胞核内AgNORs颗粒数分别为:6.2,6.5,7.3,与正常对照组细胞比较均无显著差异(P>0.05);流式细胞仪检测结果显示各组均未见明显细胞凋亡。结论:短时间电场作用对hRPE细胞的正常细胞活力及分裂无明显影响,提示该条件下的电场作用可能应用于促进RPE细胞修复的研究。

Effects of an electric field on biological characters of human retinal pigment epithelial cells

AIM:To investigate the effects of an electric field(EF)on the viability and division of human retinal pigment epithelial(hRPE)cells.· METHODS:Cultured hRPE cells were exposed to EF of 8 V/cm for 3 hours and images of the cells were obtained with an image analyzer.After that,the cells were continuously cultured and observed for an additional 12 hours.The viability and division of the cells were evaluated by typan blue and AgNORs staining,and apoptosis of the cells was determined by flow cytometry.The cells without exposing to EF served as control group.· RESULTS:In the applied direct current EF of 8 V/cm,hRPE cells changed their shapes to be elongated and oriented with their long axes perpendicular to the vector of the field.After stopped the exposure of EF,we observed for 12 hours.The cells returned to the normal shape and random distribution in the culture medium.Typan blue staining showed no differences between the cells exposed to EF and those in normal control(P>0.05).The results of AgNORs staining showed that the numbers of AgNORs in the nuclei of hRPE cells at 0 hour,3 hours in EF and 12 hours after stopped EF exposure were 6.2,6.5 and 7.3 respectively.There was no difference compared with normal control(P>0.05).The results of flow cytometry showed no obvious apoptosis of cells in EF and in control group.· CONCLUSION:Our results show that a relatively short period of exposure to EF do not impair the viability and division of hRPE cells,which allows the opportunity to study the application of EF in the enhancement of wound healing.