AIM:To investigate the acetylcholinesterase(AChE)expression involved in retina pigment epithelial(RPE)apoptosis induced by higher concentrations H_2O_2.METHODS:The human retinal pigment epithelium cell line ARPE-19 was from ATCC(Rockville,MD).Cultured ARPE-19 cells were treated with H_2O_2 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 H_2O_2>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 H_2O_2 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 H_O_2 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 H_2O_2 caused the stable increase of AChE expression in RPE cells,which may indicate that AChE may be an important role in AMD.
Li CaiHong-Fei LiaoXue-Jun ZhangYi ShaoMan XuJing-Lin Yi
