title: Protective effect of clusterin on oxidative stress-induced cell death of human corneal endothelial cells.
abstract: OBJECTIVE
  To investigate the protective effect of clusterin on oxidative stress-induced cell death of human corneal endothelial cells.
  
  
  METHODS
  Human corneal endothelial cells (HCECs) were cultured according to previously published methods. With treatment of various concentrations (0-50 mM) of tert-butyl hydroperoxide (tBHP) or clusterin, reactive oxygen species (ROS) producrion was measured using an oxidationsensitive fluorescent probe and 2'7'-dichlorofluorescin diacetate (DCFH-DA). Cell viability was assayed with a Cell Counting Kit-8.
  
  
  RESULTS
  In HCECs, DCF-DA staining revealed that cells treated with a higher concentration of tBHP had higher fluorescent intensity than cells treated with clusterin, compared to control cells. Clusterin significantly inhibited tBHP-induced ROS production. Cell viability decreased with higher tBHP concentration. Cells treated with clusterin had higher viability than control cells at 5 mM tBHP. Clusterin effectively protected HCECs from ROS-induced cell death.
  
  
  CONCLUSIONS
  Our data suggest that clusterin may protect HCECs from oxidative injury-mediated cell death via inhibition of ROS production.
authors.affiliation: Myung Dong St. Mary's Eye Center, Seoul National University College of Engineering, Seoul, Korea.
authors.forename: Young Joo
authors.initials: YJ
authors.lastname: Shin
authors.forename: Jeong Hun
authors.initials: JH
authors.lastname: Kim
authors.forename: Jong Mo
authors.initials: JM
authors.lastname: Seo
authors.forename: Sang Mok
authors.initials: SM
authors.lastname: Lee
authors.forename: Joon Young
authors.initials: JY
authors.lastname: Hyon
authors.forename: Young Suk
authors.initials: YS
authors.lastname: Yu
authors.forename: Won Ryang
authors.initials: WR
authors.lastname: Wee
mesh_terms: D000328:Adult; D053148:Caspase 3 / Q000378:metabolism; D002452:Cell Count; D016923:Cell Death; D002478:Cells, Cultured; D051152:Clusterin / Q000378:metabolism*; D019610:Cytoprotection; D042783:Endothelial Cells / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology*; D004728:Endothelium, Corneal / Q000473:pathology*; D005452:Fluoresceins / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D008856:Microscopy, Fluorescence; D018384:Oxidative Stress; D011419:Propidium / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013194:Staining and Labeling
chemical_list: D051152:Clusterin; D005452:Fluoresceins; D017382:Reactive Oxygen Species; C029569:diacetyldichlorofluorescein; D011419:Propidium; D053148:Caspase 3