RESUMEN
Reactive oxygen species (ROS) are involved in neuropathic pain, a complicated condition after nerve tissue lesion. Vitamin D appears to improve symptoms of pain and exhibits antioxidant properties. We investigated the effects of oral administration of vitamin D3, the active form of vitamin D, on nociception, the sciatic functional index (SFI), and spinal cord pro-oxidant and antioxidant markers in rats with chronic constriction injury (CCI) of the sciatic nerve, a model of neuropathic pain. Vitamin D3 (500 IU/kg per day) attenuated the CCI-induced decrease in mechanical withdrawal threshold and thermal withdrawal latency (indicators of antinociception) and SFI. The vitamin prevented increased lipid hydroperoxide levels in injured sciatic nerve without change to total antioxidant capacity (TAC). Vitamin D3 prevented increased lipid hydroperoxide, superoxide anion generation (SAG), and hydrogen peroxide (H2O2) levels in the spinal cord, which were found in rats without treatment at 7 and 28 days post-CCI. A significant negative correlation was found between mechanical threshold and SAG and between mechanical threshold and H2O2 at day 7. Vitamin D3 also prevented decreased spinal cord total thiols content. There was an increase in TAC in the spinal cord of vitamin-treated CCI rats, compared to CCI rats without treatment only at 28 days. No significant changes were found in body weight and blood parameters of hepatic and renal function. These findings demonstrated, for first time, that vitamin D modulated pro-oxidant and antioxidant markers in the spinal cord. Since antinociception occurred in parallel with oxidative changes in the spinal cord, the oxidative changes may have contributed to vitamin D-induced antinociception.
Asunto(s)
Animales , Ratas , Neuralgia/tratamiento farmacológico , Antioxidantes , Nervio Ciático , Médula Espinal , Vitamina D , Vitaminas , Especies Reactivas de Oxígeno , Ratas Wistar , Nocicepción , Peróxido de Hidrógeno , Hiperalgesia/tratamiento farmacológicoRESUMEN
Abstract Sciatic nerve transection (SNT), a model for studying neuropathic pain, mimics the clinical symptoms of "phantom limb", a pain condition that arises in humans after amputation or transverse spinal lesions. In some vertebrate tissues, this condition decreases acetylcholinesterase (AChE) activity, the enzyme responsible for fast hydrolysis of released acetylcholine in cholinergic synapses. In spinal cord of frog Rana pipiens, this enzyme's activity was not significantly changed in the first days following ventral root transection, another model for studying neuropathic pain. An answerable question is whether SNT decreases AChE activity in spinal cord of frog Lithobates catesbeianus, a species that has been used as a model for studying SNT-induced neuropathic pain. Since each animal model has been created with a specific methodology, and the findings tend to vary widely with slight changes in the method used to induce pain, our study assessed AChE activity 3 and 10 days after complete SNT in lumbosacral spinal cord of adult male bullfrog Lithobates catesbeianus. Because there are time scale differences of motor endplate maturation in rat skeletal muscles, our study also measured the AChE activity in bullfrog tibial posticus (a postural muscle) and gastrocnemius (a typical skeletal muscle that is frequently used to study the motor system) muscles. AChE activity did not show significant changes 3 and 10 days following SNT in spinal cord. Also, no significant change occurred in AChE activity in tibial posticus and gastrocnemius muscles at day 3. However, a significant decrease was found at day 10, with reductions of 18% and 20% in tibial posticus and gastrocnemius, respectively. At present we cannot explain this change in AChE activity. While temporally different, the direction of the change was similar to that described for rats. This similarity indicates that bullfrog is a valid model for investigating AChE activity following SNT.
Resumo A transecção do nervo isquiático (SNT), um modelo para estudar dor neuropática, simula os sintomas clínicos do "membro fantasma", uma condição dolorosa que ocorre nos humanos após amputação ou secção completa da medula espinal. Essa condição muda a atividade da acetilcolinesterase (AChE), a enzima responsável pela rápida hidrólise da acetilcolina liberada nas sinapses colinérgicas, em alguns tecidos de vertebrados. Em medula espinal de rã Rana pipiens, a atividade da AChE não foi significativamente alterada nos primeiros dias após a secção da raiz ventral, outro modelo para o estudo da dor neuropática. Uma questão ainda não respondida é se a SNT diminui a atividade da AChE na medula espinal de rã Lithobates catesbeianus, uma espécie que vem sendo usada como modelo em estudos da dor neuropática induzida por SNT. Como cada modelo animal é criado a partir de metodologia específica, e seus resultados tendem a variar com pequenas mudanças na metodologia de indução da dor, o presente estudo avaliou a atividade da AChE em medula espinal lombossacral de rã-touro Lithobates catesbeianus, adultos, machos, 3 e 10 dias após a completa SNT. Como há diferenças temporais na maturação de placas motoras em músculos esqueléticos de ratos, nosso estudo ainda demonstrou, na rã-touro, os efeitos da SNT sobre a atividade da AChE nos músculos esqueléticos tibial posticus, um músculo postural, e gastrocnêmio, um músculo frequentemente usado em estudos do sistema motor. A atividade da AChE não mudou significativamente na medula espinal aos 3 e 10 dias após a SNT. Nos músculos, a atividade não alterou significativamente aos 3 dias após a lesão, mas reduziu de forma significativa aos 10 dias após a SNT. Aos 10 dias, a diminuição foi 18% no músculo tibial posticus e 20% no gastrocnêmio. No momento, nós não temos explicação para essa mudança na atividade da AChE. Embora temporalmente diferente, o sentido da mudança é similar ao que é descrito em ratos. Esta similaridade torna a rã-touro um modelo válido para se estudar questões ainda não respondidas da SNT sobre a AChE.
Asunto(s)
Animales , Acetilcolinesterasa/metabolismo , Nervio Ciático/enzimología , Nervio Ciático/fisiopatología , Nervio Ciático/lesiones , Médula Espinal/fisiología , Músculo Esquelético/inervación , Rana catesbeianaRESUMEN
The effects of in vivo chronic treatment and in vitro addition of imipramine, a tricyclic antidepressant, or fluoxetine, a selective serotonin reuptake inhibitor, on the cortical membrane-bound Na+,K+-ATPase activity were studied. Adult Wistar rats received daily intraperitoneal injections of 10 mg/kg of imipramine or fluoxetine for 14 days. Twelve hours after the last injection rats were decapitated and synaptic plasma membranes (SPM) from cerebral cortex were prepared to determine Na+,K+-ATPase activity. There was a significant decrease (10 percent) in enzyme activity after imipramine but fluoxetine treatment caused a significant increase (27 percent) in Na+,K+-ATPase activity compared to control (P<0.05, ANOVA; N = 7 for each group). When assayed in vitro, the addition of both drugs to SPM of naive rats caused a dose-dependent decrease in enzyme activity, with the maximal inhibition (60-80 percent) occurring at 0.5 mM. We suggest that a) imipramine might decrease Na+,K+-ATPase activity by altering membrane fluidity, as previously proposed, and b) stimulation of this enzyme might contribute to the therapeutic efficacy of fluoxetine, since brain Na+,K+-ATPase activity is decreased in bipolar patients
Asunto(s)
Animales , Ratas , Antidepresivos , Corteza Cerebral , Fluoxetina , Imipramina , ATPasa Intercambiadora de Sodio-Potasio , Membranas Sinápticas , Antidepresivos Tricíclicos , Corteza Cerebral , Ratas Wistar , Inhibidores Selectivos de la Recaptación de Serotonina , ATPasa Intercambiadora de Sodio-Potasio , Membranas SinápticasRESUMEN
2-Hydroxybutyric acid appears at high concentrations in situations related to deficient energy metabolism (e.g., birth asphyxia) and also in inherited metabolic diseases affecting the central nervous system during neonatal development, such as "cerebral" lactic acidosis, glutaric aciduria type II, dihydrolipoyl dehydrogenase (E3) deficiency, and propionic acidemia. The present study was carried out to determine the effect of 2-hydroxybutyric acid at various concentrations (1-10 mM) on CO2 production and lipid synthesis from labeled substrates in cerebral cortex of 30-day-old Wistar rats in vitro. CO2 production was significantly inhibited (30-70 percent) by 2-hydroxybutyric acid in cerebral cortex prisms, in total homogenates and in the mitochondrial fraction. We also demonstrated a significant inhibition of lipid synthesis (20-45 percent) in cerebral cortex prisms and total homogenates in the presence of 2-hydroxybutyric acid. However, no inhibition of lipid synthesis occurred in homogenates free of nuclei and mitochondria. The results indicate an impairment of mitochondrial energy metabolism caused by 2-hydroxybutyric acid, a fact that may secondarily lead to reduction of lipid synthesis. It is possible that these findings may be associated with the neuropathophysiology of the situations where 2-hydroxybutyric acid is accumulated
Asunto(s)
Animales , Ratas , Dióxido de Carbono/metabolismo , Corteza Cerebral/efectos de los fármacos , Metabolismo Energético , Hidroxibutiratos/farmacología , Lípidos/síntesis química , Análisis de Varianza , Hidroxibutiratos/química , Mitocondrias/metabolismo , Ratas WistarRESUMEN
Levels of methylmalonic acid (MMA) comparable to those of human methylmalonic acidemia were achieved in blood (2-2.5 mmol/l) and brain (1.35 æmol/g) of rats by administering buffered MMA, pH 7.4, subcutaneously twice a day from the 5th to the 28th day of life. MMA doses ranged from 0.76 to 1.67 æmol/g as a function of animal age. Control rats were treated with saline in the same volumes. The animals were sacrificed by decapitation on the 28th day of age. Blood was taken and the brain was rapidly removed. Medulla, pons, the olfactory lobes and cerebellum were discarded and the rest of the brain ("cerebrum") was isolated. Body and "cerebrum" weight were measured, as well as the cholesterol and triglyceride concentrations in blood and the content of myelin, total lipids, and the concentrations of the lipid fractions (cholesterol, glycerolipids, phospholipids and ganglioside N-acetylneuraminic acid (ganglioside-NANA)) in the "cerebrum". Chronic MMA administration had no effect on body or "cerebrum" weight, suggesting that the metabolites per se neither affect the appetite of the rats nor cause malnutrition. In contrast, MMA caused a significant reduction of plasma triglycerides, but not of plasma cholesterol levels. A significant diminution of myelin content and of ganglioside-NANA concentration was also observed in the "cerebrum". We propose that the reduction of myelin content and ganglioside-NANA caused by MMA may be related to the delayed myelination/cerebral atrophy and neurological dysfunction found in methylmalonic acidemic children
Asunto(s)
Encéfalo , Lípidos , Ácido Metilmalónico/administración & dosificación , Proteínas de la Mielina , Vaina de Mielina , Ácido N-Acetilneuramínico , Animales Recién Nacidos , Colesterol , Gangliósidos , Ácido Metilmalónico/farmacología , Fosfolípidos/análisis , Ratas Wistar , Triglicéridos/sangreRESUMEN
The effects of transient forebrain ischemia, reperfusion and ischemic preconditioning on rat blood platelet ATP diphosphohydrolase and 5'-nucleotidase activities were evaluated. Adult Wistar rats were submitted to 2 or 10 min of single ischemic episodes, or to 10 min of ischemia 1 day after a 2-min ischemic episode (ischemic preconditioning) by the four-vessel occlusion method. Rats submitted to single ischemic insults were reperfused for 60 min and for 1, 2, 5, 10 and 30 days after ischemia; preconditioned rats were reperfused for 60 min 1 and 2 days after the long ischemic episode. Brain ischemia (2 or 10 min) inhibited ATP and ADP hydrolysis by platelet ATP diphosphohydrolase. On the other hand, AMP hydrolysis by 5'-nucleotidase was increased after 2, but not 10, min of ischemia. Ischemic preconditioning followed by 10 min of ischemia caused activation of both enzymes. Variable periods of reperfusion distinctly affected each experimental group. Enzyme activities returned to control levels in the 2-min group. However, the decrease in ATP diphosphohydrolase activity was maintained up to 30 days of reperfusion after 10-min ischemia. 5'-Nucleotidase activity was decreased 60 min and 1 day following 10-min ischemia; interestingly, enzymatic activity was increased after 2 and 5 days of reperfusion, and returned to control levels after 10 days. Ischemic preconditioning cancelled the effects of 10-min ischemia on the enzymatic activities. These results indicate that brain ischemia and ischemic preconditioning induce peripheral effects on ecto-enzymes from rat platelets involved in nucleotide metabolism. Thus, ATP, ADP and AMP degradation and probably the generation of adenosine in the circulation may be altered, leading to regulation of microthrombus formation since ADP aggregates platelets and adenosine is an inhibitor of platelet aggregation
Asunto(s)
Animales , Ratas , Masculino , 5'-Nucleotidasa/metabolismo , Apirasa/metabolismo , Plaquetas/química , Isquemia Encefálica/enzimología , Análisis de Varianza , Precondicionamiento Isquémico , Ratas Wistar , Factores de TiempoRESUMEN
ATP diphosphohydrolase (apyrase)(EC3.6.1.5) activity was measured in synaptosomes from cerebral cortex of Wistar rats of both sexes subjected to experimental phenylketonuria, i.e., chemical hyperphenylaninemia induced by subcutaneous administration of 5.2 µmol phenylalanine/g body weight (twice a day) plus 0.9 µmol p-chlorophenylalanine/g body weight (once a day). ATP diphosphohydrolase specific activity (nmol Pi min-1 mg protein-1) of synaptosomes was significantly decreased compared to controls for both ATp (from 147.6 to 129.9) and ADP (from 70.2 to 63.1) hydrolysis one hour after single administration of the drugs to 35-day old rats. Chronic treatment was performed from the 6th to the 28th postpartum day. The enzyme specific activity of synaptosomes was measured one week after the last administration of the drugs and was significantly reduced compared to controls for both ATP (from 164.1 to 150.2) and ADP (from 76.3 to 62.1) hydrolysis. The in vitro effects of the drugs on the synaptosome enzyme specific activity were also investigated. Phenylalnine alone or associated with p-chlorophenylalanine significantly reduced enzyme specific activity for both ATP (from 150.2 to 136.0) and ADP (from 70.5 to 59.3) nucleotides as substrates. Since ATP diphosphohrolase seems to play an important role in neurotransmission, these findings may be related to the neurological dysfunction characteristic of human phenylketonuria