Acetylcholinesterase function in apoptotic retina pigment epithelial cells induced by H_2O_2  

作　　者：Li Cai Hong-Fei Liao Xue-Jun Zhang Yi Shao Man Xu Jing-Lin Yi 

机构地区：[1]Jiangxi Research Institute of Ophthalmology and Visual Sciences [2]Laboratory of Molecular Cell Biology,Institute ofBiochemistry and Cell Biology,Shanghai Institutes forBiological Sciences [3]Department of Ophthalmology,the First Affiliated Hospitalof Nanchang University

出　　处：《International Journal of Ophthalmology(English edition)》2013年第6期772-777,共6页国际眼科杂志（英文版）

基　　金：National Natural Science Foundation of China(No.31071213,81101479,30971481,81260148,81271425 and 81160118);Clinical Medicine Research Special-purpose Foundation of China(No.L2012052);Natural Science Foundation of Jiangxi Province,China(No.20114BAB215029);Technology Foundation of Jiangxi Province,China(No 20111BBG70026-2);Health Department Science and Technology Foundation of Jiangxi Province,China(No.20121026);Education Department Youth Scientific Research Foundation of Jiangxi Province,China(No.GJJ12158)

摘　　要：AIM:To investigate the acetylcholinesterase（AChE）expression involved in retina pigment epithelial（RPE）apoptosis induced by higher concentrations H2O2.METHODS:The human retinal pigment epithelium cell line ARPE-19 was from ATCC（Rockville,MD）.Cultured ARPE-19 cells were treated with H2O2 at 0,250,500,1000,2 000μmol/L and cell viability was measured with MTT assay.AChE expression and DNA fragments were analyzed by immunocytochemistry,TUNEL and PARP-1Western blotting.RESULTS:Immunofluorescence detected AChE exist in the normal human retinal tissue.When H2O2】500μmol/L,AChE expression showed an increase after 2h,and this concentration was selected for the present study.RPE cell was induced with 1 000μmoI/L H2O2 for 2h,compared to the control group,cell activity decline detected by MTT,AChE and PARP-1 protein expression was significantly increased detected by Western blotting.AChE immunofluorescence staining was positive in RPE cell after HO2 incubate 2h.In addition,pretreatment with100|jmol/L epigallocatechin gallate（EGCG）,cell viability increased from 31.20%±3.90%to 70.23%±12.96%.CONCLUSION:AChE is weakly expressed in normal human RPE cells.Stimulation with H2O2 caused the stable increase of AChE expression in RPE cells,which may indicate that AChE may be an important role in AMD.AIM:To investigate the acetylcholinesterase（AChE）expression involved in retina pigment epithelial（RPE）apoptosis induced by higher concentrations H2O2.METHODS:The human retinal pigment epithelium cell line ARPE-19 was from ATCC（Rockville,MD）.Cultured ARPE-19 cells were treated with H2O2 at 0,250,500,1000,2 000μmol/L and cell viability was measured with MTT assay.AChE expression and DNA fragments were analyzed by immunocytochemistry,TUNEL and PARP-1Western blotting.RESULTS:Immunofluorescence detected AChE exist in the normal human retinal tissue.When H2O2>500μmol/L,AChE expression showed an increase after 2h,and this concentration was selected for the present study.RPE cell was induced with 1 000μmoI/L H2O2 for 2h,compared to the control group,cell activity decline detected by MTT,AChE and PARP-1 protein expression was significantly increased detected by Western blotting.AChE immunofluorescence staining was positive in RPE cell after HO2 incubate 2h.In addition,pretreatment with100|jmol/L epigallocatechin gallate（EGCG）,cell viability increased from 31.20%±3.90%to 70.23%±12.96%.CONCLUSION:AChE is weakly expressed in normal human RPE cells.Stimulation with H2O2 caused the stable increase of AChE expression in RPE cells,which may indicate that AChE may be an important role in AMD.

关 键 词：ACETYLCHOLINESTERASE retina pigment epithelial cells oxidative stress age-related macular degeneration 

分 类 号：R774[医药卫生—眼科]