电场对人视网膜色素上皮细胞增生的影响  

Effects of electric field on proliferation of human retinal pigment epithelial cells

作　　者：韩静[1] 闫小龙[2] 惠延年[3] 韩泉洪[4] 

机构地区：[1]第四军医大学唐都医院眼科,陕西省西安市710038 [2]第四军医大学唐都医院胸外科,陕西省西安市710038 [3]第四军医大学西京医院眼科,全军眼科研究所,陕西省西安市710032 [4]天津市眼科医院,300070

出　　处：《眼科新进展》2010年第2期101-104,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30670510);德国洪堡基金会(Alexander von Humboldt Founda-tion)仪器设备捐赠基金资助(编号:V-8151 /02085)~~

摘　　要：目的观察一定条件下电场对人视网膜色素上皮(human retinal pigment epitheli-um,hRPE)细胞增生的影响及相关机制。方法hRPE细胞置于电场强度为6V.cm-1的电场中连续作用12h。观察未受电场作用的正常对照组及电场作用12h的实验组细胞,显微摄像系统记录不同时间点细胞图像并计数细胞数,检测细胞密度和细胞的生长速度,观测指标包括:细胞密度,以每平方厘米细胞数表示;细胞生长速度,计算公式为:(Ni-N0)/N0×100%(Ni为12h细胞数;N0为0h细胞数);采用流式细胞仪测定细胞周期,计算细胞增生指数:(NS+NG2/M)/N(NS为S期细胞数;NG2/M为G2/M期细胞数;N为细胞总数);Western blotting检测细胞cyclinE蛋白的表达。结果对照组hRPE细胞在12h的观察期间内细胞密度缓慢上升,到12h时细胞密度由95×103cm-2增至130×103cm-2;而实验组在6V.cm-1电场作用下hRPE细胞数量逐渐上升,12h后细胞密度上升至150×103cm-2。实验组hRPE细胞生长速度为50.02%,较对照组(36.84%)明显升高(P<0.05);流式细胞仪检测结果显示电场作用12h后G0/G1期细胞比例下降,进入S期和G2/M期的细胞比例增多,对照组细胞增生指数为38.43%,实验组升至55.02%(P<0.05);Western blotting结果进一步显示,电场作用12h后hRPE细胞cyclinE蛋白表达水平实验组(比值为0.81±1.22)与对照组(比值为0.47±1.38)相比显著升高(P<0.05)。结论一定条件下的电场作用能够通过上调细胞周期蛋白的表达促进hRPE细胞的增生。Objective To investigate the effects and mechanisms of electric field（EF） on proliferation of human retinal pigment epithelial（hRPE） cells.Methods Cultured hRPE cells were continuously exposed to EF with 6 V·cm^-1 EF strength for 12 hours.Cells in normal control group （without EF treatment） and experimental group（treated with EF for 12 hours） were observed.Cell images were recorded and cell numbers were calculated at different times with micro-image system.Cell density （cell number·cm^-2） was assessed and cell growth rates were calculated as [（Ni-N0）/N0]×100%（Ni：cell numbers at 12 hours;N0：cell numbers at 0 hour）.Flow cytometry was used to measure cell cycle and calculate cellular proliferation index （NS＋NG2/M）/N（NS：cell number at S phase;NG2/M：cell number at G2/M phase;N：total cell number）,and the expression of cyclin E in hRPE cells was determined by Western blotting.Results In normal control group,cell density was increased slowly from 95×10^3 cm^-2 to 130×10^3 cm^-2 in observing time for 12 hours;In experimental group,number of hRPE cells was increased gradually and cell denstiy increased to 150×10^3 cm^-2 under 6 V·cm^-1 EF strength.Growth velocity was 50.02% in experimental group higher than 36.84% in normal control group（P〈0.05）.Flow cytometry showed that the percentage of cell population increased in S and G2/M phases,decreased significantly in G0/G1 phase under EF exposure for 12 hours,and the proliferation index increased from 38.43% in normal control group to 55.02% in experimental group（P〈0.05）.Western blotting analysis showed that EF exposure for 12 hours induced a significant increase in the ratio of cyclin E expression in hRPE cells from 0.47±1.38 in normal control group to 0.81±1.22 in experimental group（P〈0.05）.Conclusion EF can promote the proliferation of hRPE cells by up-regulating the expression of cell cycle related proteins under certain conditions.

关 键 词：电场 视网膜色素上皮细胞 细胞增生 

分 类 号：R774[医药卫生—眼科]