title: Acetylcholinesterase function in apoptotic retina pigment epithelial cells induced by H2O2.
abstract: OBJECTIVE
  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, 1 000, 2 000µmol/L and cell viability was measured with MTT assay. AChE expression and DNA fragments were analyzed by immunocytochemistry, TUNEL and PARP-1 Western 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 000µmol/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 H2O2 incubate 2h. In addition, pretreatment with 100µmol/L epigallocatechin gallate (EGCG), cell viability increased from 31.20%±3.90% to 70.23%±12.96%.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Jiangxi Research Institute of Ophthalmology and Visual Sciences, Nanchang 330006, Jiangxi Province, China.
authors.forename: Li
authors.initials: L
authors.lastname: Cai
authors.affiliation: Jiangxi Research Institute of Ophthalmology and Visual Sciences, Nanchang 330006, Jiangxi Province, China.
authors.forename: Hong-Fei
authors.initials: HF
authors.lastname: Liao
authors.affiliation: Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Shanghai 200000, China.
authors.forename: Xue-Jun
authors.initials: XJ
authors.lastname: Zhang
authors.affiliation: Department of Ophthalmology, the First Affiliated Hospital of Nanchang University, Nanchang 330006, Jiangxi Province, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Shao
authors.affiliation: Jiangxi Research Institute of Ophthalmology and Visual Sciences, Nanchang 330006, Jiangxi Province, China.
authors.forename: Man
authors.initials: M
authors.lastname: Xu
authors.affiliation: Jiangxi Research Institute of Ophthalmology and Visual Sciences, Nanchang 330006, Jiangxi Province, China.
authors.forename: Jing-Lin
authors.initials: JL
authors.lastname: Yi
keywords: acetylcholinesterase; age-related macular degeneration; oxidative stress; retina pigment epithelial cells