title: Resveratrol prevention of oxidative stress damage to lens epithelial cell cultures is mediated by forkhead box O activity.
abstract: OBJECTIVE
  To evaluate the potential role that FoxO transcription factors play in modulating resveratrol's protective effects against oxidative stress in lens epithelial cells.
  
  
  METHODS
  Primary human or porcine lens epithelial cells (LECs) were treated with resveratrol (RES) 25 μM and incubated under either physiologic (5%) or chronic hyperoxic (40%) oxygen conditions. Acute oxidative stress was applied using 600 μM H(2)O(2). Changes in expression of FoxO1A, FoxO3A, and FoxO4 were analyzed. The production of intracellular reactive oxygen species (iROS), SA-β-galactosidase (SA-β-gal) activity, and autofluorescence (AF) was assessed by flow cytometry. SiRNAs of FoxO1A, FoxO3A, and FoxO4 were used to study the roles that these transcription factors play in resveratrol's protective effects against cell death induced by oxidative stress.
  
  
  RESULTS
  RES incubation under 40% oxygen increased the expression of FoxO1A, FoxO3A, and FoxO4. RES also increases mitochondrial membrane potential under 5% and/or 40% O(2) conditions and significantly decreased iROS, SA-β-gal, and AF normally induced by hyperoxic conditions. While RES had a mild pro-apoptotic effect in nonstressed cells, it significantly prevented apoptosis induced by H(2)O(2) stress. SiRNA inhibition of FoxO1A, FoxO3A, and FoxO4 not only led to loss of the anti-apoptotic effects of RES in stressed cells but actually exhibited a mild pro-apoptotic effect.
  
  
  CONCLUSIONS
  RES exerts a protective effect against oxidative damage in LEC cultures. The levels of expression of FoxO1A, FoxO3A, and FoxO4 appear to play a central role in determining the pro- or anti-apoptotic effects of RES. This has implications for future studies on oxidative stress-related lenticular disorders such as cataract formation.
authors.affiliation: Department of Ophthalmology, Duke Eye Center, Duke University Medical Center, Durham, North Carolina 27710, USA.
authors.forename: Guorong
authors.initials: G
authors.lastname: Li
authors.forename: Coralia
authors.initials: C
authors.lastname: Luna
authors.forename: Iris D
authors.initials: ID
authors.lastname: Navarro
authors.forename: David L
authors.initials: DL
authors.lastname: Epstein
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.forename: Pedro
authors.initials: P
authors.lastname: Gonzalez
authors.forename: Pratap
authors.initials: P
authors.lastname: Challa
mesh_terms: D020533:Angiogenesis Inhibitors; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002386:Cataract / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D005434:Flow Cytometry; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007908:Lens, Crystalline / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D053078:Membrane Potential, Mitochondrial; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology*; D013552:Swine
chemical_list: D020533:Angiogenesis Inhibitors; D000975:Antioxidants; D051858:Forkhead Transcription Factors; D013267:Stilbenes; D012313:RNA; D000077185:Resveratrol