title: Docosahexaenoic acid (DHA) has neuroprotective effects against oxidative stress in retinal ganglion cells.
abstract: We examined the radical-scavenging activity of docosahexaenoic acid (DHA) and its effects on the neuronal cell death induced by oxidative or hypoxic stress in cultured retinal ganglion cells (RGC-5, a rat ganglion cell-line transformed using E1A virus). The radical-scavenging activity [hydrogen peroxide (H(2)O(2)), superoxide anion (O(2)*(-)), and hydroxyl radical (*OH)] of DHA in RGC-5 cells was measured using the ROS-sensitive probes CM-H(2)DCFDA and APF. DHA concentration-dependently scavenged the intracellular radical productions induced by H(2)O(2) radical, O(2)*(-), and *OH (minimum effective DHA concentrations 0.1, 10 and 100 microM, respectively). Cell damage was induced by H(2)O(2), oxygen-glucose deprivation (OGD), or tunicamycin (an endoplasmic reticulum-stress inducer), and cell viability was assessed by Hoechst 33342 nuclear staining or by the tetrazolium salt (WST-8) cell-viability assay. H(2)O(2) (0.3 mM for 24 h), 4-h OGD exposure followed by 18-h reoxygenation, or tunicamycin at 2 microg/ml for 24 h induced apoptotic cell death accompanied by nuclear condensation and/or fragmentation, and each maneuver decreased cell viability. Treatment with DHA at 0.1 and 1 microM significantly inhibited the decrease in cell viability induced by H(2)O(2). Treatment with DHA at 0.1, 1, or 10 microM significantly inhibited the decrease in cell viability induced by OGD/reoxygenation exposure. However, DHA (0.1 to 10 microM) had no effect on the decrease in cell viability induced by tunicamycin. These results indicate that DHA may be protective against oxidative or hypoxic stress-induced cell damage in retinal ganglion cells.
authors.affiliation: Department of Biofunctional Evaluation, Molecular Pharmacology, Gifu Pharmaceutical University, 5-6-1 Mitahora-higashi, Gifu 502-8585, Japan.
authors.forename: Masamitsu
authors.initials: M
authors.lastname: Shimazawa
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Nakajima
authors.forename: Yukihiko
authors.initials: Y
authors.lastname: Mashima
authors.forename: Hideaki
authors.initials: H
authors.lastname: Hara
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D019610:Cytoprotection / Q000187:drug effects* / Q000502:physiology; D004281:Docosahexaenoic Acids / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D016166:Free Radical Scavengers / Q000378:metabolism / Q000494:pharmacology; D005609:Free Radicals / Q000037:antagonists & inhibitors / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors; D000860:Hypoxia / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D007511:Ischemia / Q000188:drug therapy / Q000378:metabolism / Q000503:physiopathology; D018696:Neuroprotective Agents / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology; D051381:Rats; D012165:Retinal Ganglion Cells / Q000187:drug effects* / Q000378:metabolism; D013481:Superoxides / Q000037:antagonists & inhibitors; D014415:Tunicamycin / Q000494:pharmacology
chemical_list: D016166:Free Radical Scavengers; D005609:Free Radicals; D018696:Neuroprotective Agents; D013481:Superoxides; D014415:Tunicamycin; D004281:Docosahexaenoic Acids; D006861:Hydrogen Peroxide