title: Nutraceutical with Resveratrol and Omega-3 Fatty Acids Induces Autophagy in ARPE-19 Cells.
abstract: Impaired autophagic and proteasomal cleansing have been documented in aged retinal pigment epithelial (RPE) cells and age-related macular degeneration (AMD). Omega-3 fatty acids and resveratrol have many positive homeostatic effects in RPE cells. In this work, ARPE-19 cells were treated with 288 ng of Resvega, containing 30 mg of trans resveratrol and 665 mg of omega-3 fatty acids, among other nutrients, with proteasome inhibitor MG-132 or autophagy inhibitor bafilomycin A1 up to 48 h. Autophagy markers p62/SQSTM1 (p62) and LC3 (microtubule-associated protein 1A/1B-light chain 3) were analyzed by Western blotting. Fluorescence microscopy with mCherry-GFP-LC3 plasmid was applied to study the autophagy flux, and cytoprotective effects were investigated with colorimetric MTT and LDH assays. Resvega induced autophagy by showing increased autolysosome formation and autophagy flux, and the change in the p62 and LC3 protein levels further confirmed the fluorescent microscopy results. Moreover, Resvega provided a clear cytoprotection under proteasome inhibition. These findings highlight the potential of the nutraceuticals containing resveratrol, omega-3 fatty acids and other nutrients in the prevention of ARPE-19 cell damage.
authors.affiliation: Department of Ophthalmology, University of Eastern Finland, Kuopio 70211, Finland. ali.koskela@uef.fi.
authors.forename: Ali
authors.initials: A
authors.lastname: Koskela
authors.affiliation: School of Pharmacy, University of Eastern Finland, Kuopio 70211, Finland. mika.reinisalo@uef.fi.
authors.forename: Mika
authors.initials: M
authors.lastname: Reinisalo
authors.affiliation: Stem Cells and Eye Research Laboratory, Department of Ophthalmology, Faculty of Medicine, University of Szeged, Szeged 6720, Hungary. goran.petrovski@medisin.uio.no.|Centre of Eye Research, Department of Ophthalmology, Oslo University Hospital, University of Oslo, Oslo 0450, Norway. goran.petrovski@medisin.uio.no.
authors.forename: Goran
authors.initials: G
authors.lastname: Petrovski
authors.affiliation: Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. debasish@jhmi.edu.
authors.forename: Debasish
authors.initials: D
authors.lastname: Sinha
authors.affiliation: Laboratoires Théa, Clermond-Ferrand cedex 63 000, France. c.olmiere@laboratoires-thea.fr.
authors.forename: Céline
authors.initials: C
authors.lastname: Olmiere
authors.affiliation: Department of Biology, University of Eastern Finland, Kuopio 70211, Finland. reijo.karjalainen@uef.fi.
authors.forename: Reijo
authors.initials: R
authors.lastname: Karjalainen
authors.affiliation: Department of Ophthalmology, University of Eastern Finland, Kuopio 70211, Finland. kai.kaarniranta@uef.fi.|Department of Ophthalmology, Kuopio University Hospital, Kuopio 70029, Finland. kai.kaarniranta@uef.fi.
authors.forename: Kai
authors.initials: K
authors.lastname: Kaarniranta
mesh_terms: D001343:Autophagy / Q000187:drug effects*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D019587:Dietary Supplements / Q000032:analysis*; D004791:Enzyme Inhibitors / Q000008:administration & dosage / Q000494:pharmacology; D015525:Fatty Acids, Omega-3 / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D018942:Macrolides / Q000008:administration & dosage / Q000494:pharmacology; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D000077185:Resveratrol; D013267:Stilbenes / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; D015525:Fatty Acids, Omega-3; D007976:Leupeptins; C477012:MAP1LC3A protein, human; D018942:Macrolides; D008869:Microtubule-Associated Proteins; C118585:P62 protein, human; D016601:RNA-Binding Proteins; D013267:Stilbenes; C040929:bafilomycin A1; D000077185:Resveratrol; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde
keywords: LC3; autophagy; omega-3; p62/SQSTM1; proteasome; resveratrol