title: Sulforaphane decreases endothelial cell apoptosis in fuchs endothelial corneal dystrophy: a novel treatment.
abstract: OBJECTIVE
  Fuchs endothelial corneal dystrophy (FECD) is an oxidative stress disorder that leads to age-related and gradual loss of corneal endothelial cells resulting in corneal edema and loss of vision. To date, other than surgical intervention, there are no treatment options for patients with FECD. We have shown that in FECD, there is a deficiency in nuclear factor erythroid 2-related factor 2 (Nrf2)-regulated antioxidant defense due to decreased Nrf2 nuclear translocation and activation of antioxidant response element (ARE). In this study, we used sulforaphane (SFN) and D3T to investigate a strategy of targeting Nrf2-ARE in FECD.
  
  
  METHODS
  FECD and normal ex vivo corneas and human corneal endothelial cell lines were pretreated with SFN or D3T and exposed to oxidative stress with tert-Butyl hydroperoxide (tBHP). Apoptosis was detected with TUNEL. Cellular localization of Nrf2 and p53 was assessed by immunohistochemistry. Effect of SFN was determined by using DCFDA assay, Western blot and real-time PCR.
  
  
  RESULTS
  After pretreatment with SFN, oxidative stress was induced with tBHP. In ex vivo FECD specimens, SFN decreased CEC apoptosis by 55% in unstressed group and by 43% in tBHP-treated specimens. SFN enhanced nuclear translocation of Nrf2 in FECD specimens and decreased p53 staining under oxidative stress. Pretreatment with SFN enhanced cell viability by decreasing intracellular reactive oxygen species production. Upregulation of Nrf2 levels led to increased synthesis of DJ-1, heme oxygenase 1, and nicotinamide adenine dinucleotide quinone oxidoreductase-1. SFN significantly upregulated major ARE-dependent antioxidants and ameliorated oxidative stress-induced apoptosis in FECD.
  
  
  CONCLUSIONS
  Our results suggest that targeting Nrf2-ARE pathway may arrest degenerative cell loss seen in FECD.
authors.affiliation: Schepens Eye Research Institute, Massachusetts Eye and Ear, Department of Ophthalmology, Harvard Medical School, Boston, Massachusetts.
authors.forename: Alireza
authors.initials: A
authors.lastname: Ziaei
authors.forename: Thore
authors.initials: T
authors.lastname: Schmedt
authors.forename: Yuming
authors.initials: Y
authors.lastname: Chen
authors.forename: Ula V
authors.initials: UV
authors.lastname: Jurkunas
mesh_terms: D000368:Aged; D016588:Anticarcinogenic Agents / Q000627:therapeutic use; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D015153:Blotting, Western; D002460:Cell Line; D016039:Corneal Transplantation; D004728:Endothelium, Corneal / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D005642:Fuchs' Endothelial Dystrophy / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D006801:Humans; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D017879:Isothiocyanates / Q000627:therapeutic use*; D008297:Male; D051267:NF-E2-Related Factor 2 / Q000096:biosynthesis* / Q000235:genetics; D018384:Oxidative Stress / Q000187:drug effects* / Q000235:genetics; D012313:RNA / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D013454:Sulfoxides; D015854:Up-Regulation / Q000187:drug effects / Q000235:genetics
chemical_list: D016588:Anticarcinogenic Agents; D017879:Isothiocyanates; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D013454:Sulfoxides; D012313:RNA; C016766:sulforaphane
keywords: DJ-1; Fuchs endothelial corneal dystrophy; Hmox-1; NQO1; Nrf2; apoptosis; corneal endothelial cells; oxidative stress; reactive oxygen species; sulforaphane
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