title: Effects of antioxidant components of AREDS vitamins and zinc ions on endothelial cell activation: implications for macular degeneration.
abstract: OBJECTIVE
  To investigate whether the benefit of Age-Related Eye Disease Study (AREDS) formula multivitamins and zinc in the progression of age-related macular degeneration (AMD) may occur through inhibiting inflammatory events in the choroid.
  
  
  METHODS
  Mouse C166 endothelial cells (ECs) and, for some experiments, human retinal pigment epithelium (RPE)-choroid organ cultures were treated with AREDS multivitamin solution (MVS) or ZnCl(2). The cytotoxicity of MVS was evaluated using a lactate dehydrogenase colorimetric assay. Cell motility was assessed using a scratch assay. Macrophage adhesion to EC monolayers or ICAM-1 protein was determined after MVS and zinc treatment and with or without lipopolysaccharide (LPS). Quantitative reverse transcription PCR and Western blot analysis were used to determine the effects of MVS on the expression of proinflammatory molecules in treated and untreated cells.
  
  
  RESULTS
  AREDS MVS and zinc did not affect C166 EC viability until the 56th hour after treatment. Scratch assays showed partial inhibition of MVS and zinc on EC migration. In cell adhesion assays, MVS and zinc decreased the number of macrophages bound to EC and to ICAM-1 protein. Quantitative PCR showed that LPS increased the expression of ICAM-1 in both C166 and human RPE-choroid cultures, which was partially offset by MVS and zinc. MVS and zinc also mitigated LPS-induced ICAM-1 protein expression on Western blot analysis.
  
  
  CONCLUSIONS
  Treatment with AREDS MVS and zinc may affect both angiogenesis and endothelial-macrophage interactions. These results suggest that AREDS vitamins and zinc ions may slow the progression of AMD, in part through the attenuation of EC activation.
authors.affiliation: Department of Ophthalmology and Visual Sciences, Institute for Vision Research, The University of Iowa, Iowa City, Iowa 52242, USA.
authors.forename: Shemin
authors.initials: S
authors.lastname: Zeng
authors.forename: Jasmine
authors.initials: J
authors.lastname: Hernández
authors.forename: Robert F
authors.initials: RF
authors.lastname: Mullins
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002829:Choroid / Q000187:drug effects* / Q000276:immunology; D019587:Dietary Supplements; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000187:drug effects / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000502:physiology; D008268:Macular Degeneration / Q000188:drug therapy*; D051379:Mice; D018919:Neovascularization, Physiologic / Q000187:drug effects; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014131:Trace Elements / Q000494:pharmacology*; D014815:Vitamins / Q000494:pharmacology*; D015032:Zinc / Q000494:pharmacology*
chemical_list: D000975:Antioxidants; C090509:Icam1 protein, mouse; D014131:Trace Elements; D014815:Vitamins; D018799:Intercellular Adhesion Molecule-1; D015032:Zinc