title: Rapamycin reduces reactive oxygen species in cultured human corneal endothelial cells.
abstract: OBJECTIVE
  To investigate the protective effect of rapamycin on oxidative stress-induced cell death of human corneal endothelial cells (HCECs).
  
  
  METHODS
  HCECs were cultured according to previously published methods. With treatment of 0 mM or 5 mM of tert-butyl hydroperoxide (tBHP) with various concentrations (0, 25 and 50 nM) of rapamycin, reactive oxygen species (ROS) production was measured using an oxidation-sensitive fluorescent probe, 2'7'-dichlorofluorescin diacetate (DCFH-DA, USA) methods. Cell viability was assayed by the method of Cell Counting Kit-8 (CCK-8, Wako). The levels of cellular glutathione were also assessed enzymatically with glutathione reductase by using a commercial glutathione (GSH) assay kit (Cayman Chemical, USA).
  
  
  RESULTS
  Rapamycin reduced 2'7'-dihydrodichlorofluorescein oxidation and increased GSH in HCECs. Rapamycin significantly inhibited tBHP-induced ROS production. Cells treated with rapamycin showed higher viability compared to control at 5 mM tBHP. Rapamycin effectively protected HCECs from ROS-induced cell death through increasing intracellular GSH.
  
  
  CONCLUSIONS
  Our data suggest that rapamycin protects HCECs from oxidative injury-mediated cell death via inhibition of ROS production and enhancement of GSH.
authors: Department of Ophthalmology, Hallym University College of Medicine, Seoul, Republic of Korea. schinn@hanmail.net
authors: Young Joo
authors: YJ
authors: Shin
authors: Doo Young
authors: DY
authors: Cho
authors: Tae Young
authors: TY
authors: Chung
authors: Sang Beom
authors: SB
authors: Han
authors: Joon Young
authors: JY
authors: Hyon
authors: Won Ryang
authors: WR
authors: Wee
mesh_terms: D000328:Adult; D002452:Cell Count; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D003316:Corneal Diseases / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D004728:Endothelium, Corneal / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D008875:Middle Aged; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020123:Sirolimus / Q000494:pharmacology*
chemical_list: D007166:Immunosuppressive Agents; D017382:Reactive Oxygen Species; D020123:Sirolimus