Excitotoxic brain damage involves early peroxynitrite formation in a model of Huntington's disease in rats: protective role of iron porphyrinate 5,10,15,20-tetrakis (4-sulfonatophenyl)porphyrinate iron (III).
Oxidative/nitrosative stress is involved in NMDA receptor-mediated excitotoxic brain damage produced by the glutamate analog quinolinic acid. The purpose of this work was to study a possible role of peroxynitrite, a reactive oxygen/nitrogen species, in the course of excitotoxic events evoked by quinolinic acid in the brain. The effects of Fe(TPPS) (5,10,15,20-tetrakis (4-sulfonatophenyl)porphyrinate iron (III)), an iron porphyrinate and putative peroxynitrite decomposition catalyst, were tested on lipid peroxidation and mitochondrial function in brain synaptic vesicles exposed to quinolinic acid, as well as on peroxynitrite formation, nitric oxide synthase and superoxide dismutase activities, lipid peroxidation, caspase-3-like activation, DNA fragmentation, and GABA levels in striatal tissue from rats lesioned by quinolinic acid. Circling behavior was also evaluated. Increasing concentrations of Fe(TPPS) reduced lipid peroxidation and mitochondrial dysfunction induced by quinolinic acid (100 microM) in synaptic vesicles in a concentration-dependent manner (10-800 microM). In addition, Fe(TPPS) (10 mg/kg, i.p.) administered 2 h before the striatal lesions, prevented the formation of peroxynitrite, the increased nitric oxide synthase activity, the decreased superoxide dismutase activity and the increased lipid peroxidation induced by quinolinic acid (240 nmol/microl) 120 min after the toxin infusion. Enhanced caspase-3-like activity and DNA fragmentation were also reduced by the porphyrinate 24 h after the injection of the excitotoxin. Circling behavior from quinolinic acid-treated rats was abolished by Fe(TPPS) six days after quinolinic acid injection, while the striatal levels of GABA, measured one day later, were partially recovered. The protective effects that Fe(TPPS) exerted on quinolinic acid-induced lipid peroxidation and mitochondrial dysfunction in synaptic vesicles suggest a primary action of the porphyrinate as an antioxidant molecule. In vivo findings suggest that the early production of peroxynitrite, altogether with the enhanced risk of superoxide anion (O2*-) and nitric oxide formation (its precursors) induced by quinolinic acid in the striatum, are attenuated by Fe(TPPS) through a recovery in the basal activities of nitric oxide synthase and superoxide dismutase. The porphyrinate-mediated reduction in DNA fragmentation simultaneous to the decrease in caspase-3-like activation from quinolinic acid-lesioned rats suggests a prevention in the risk of peroxynitrite-mediated apoptotic events during the course of excitotoxic damage in the striatum. In summary, the protective effects that Fe(TPPS) exhibited both under in vitro and in vivo conditions support an active role of peroxynitrite and its precursors in the pattern of brain damage elicited by excitotoxic events in the experimental model of Huntington's disease. The neuroprotective mechanisms of Fe(TPPS) are discussed.
Pérez-De La Cruz
V
V
Laboratorio de Aminoácidos Excitadores, Instituto Nacional de Neurología y Neurocirugía Manuel Velasco Suárez, México D.F. 14269, Mexico.
González-Cortés
C
C
Galván-Arzate
S
S
Medina-Campos
O N
ON
Pérez-Severiano
F
F
Ali
S F
SF
Pedraza-Chaverrí
J
J
Santamaría
A
A
D000704:Analysis of Variance; D000818:Animals; D001522:Behavior, Animal; D015139:Blotting, Southern / Q000379:methods; D001930:Brain Injuries / Q000150:complications / Q000378:metabolism*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003342:Corpus Striatum / Q000187:drug effects / Q000378:metabolism; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006816:Huntington Disease / Q000209:etiology / Q000517:prevention & control*; D015227:Lipid Peroxidation / Q000187:drug effects / Q000502:physiology; D008297:Male; D008928:Mitochondria / Q000502:physiology; D009043:Motor Activity / Q000187:drug effects / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000378:metabolism; D030421:Peroxynitrous Acid / Q000378:metabolism*; D011166:Porphyrins / Q000627:therapeutic use*; D011805:Quinolinic Acids / Q000494:pharmacology; D011838:Radiation-Sensitizing Agents / Q000627:therapeutic use*; D051381:Rats; D017208:Rats, Wistar; D045442:Rotarod Performance Test / Q000379:methods; D013482:Superoxide Dismutase / Q000378:metabolism; D013572:Synaptic Vesicles / Q000378:metabolism; D013574:Synaptosomes / Q000502:physiology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D011166:Porphyrins; D011805:Quinolinic Acids; D011838:Radiation-Sensitizing Agents; D030421:Peroxynitrous Acid; C016136:tetraphenylporphine sulfonate; C002744:3-nitrotyrosine; D014443:Tyrosine; D005680:gamma-Aminobutyric Acid; D019001:Nitric Oxide Synthase; D013482:Superoxide Dismutase; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases