RESUMO
The chronic exposure to regular exercise training seems to improve antioxidant defense systems. However, the intense physical training imposed on elite athletes may lead to overtraining associated with oxidative stress. The purpose of the present study was to investigate the effect of different training loads and competition on oxidative stress, biochemical parameters and antioxidant enzymatic defense in handball athletes during 6-months of monitoring. Ten male elite handball athletes were recruited to the study. Blood samples were collected four times every six weeks throughout the season. During most intense periods of training and competitions there were significant changes in plasma indices of oxidative stress (increased TBARS and decreased thiols). Conversely, chronic adaptations to exercise training demonstrated a significant protective effect against oxidative stress in erythrocyte (decrease in TBARs and carbonyl group levels). Erythrocyte antioxidant enzyme activities were significantly increased, suggesting a training-induced antioxidant adaptation. Biomarkers of skeletal muscle damage were significantly increased during high-intensity training period (creatine kinase, lactate dehydrogenase and aspartate aminotransferase). No significant changes were observed in plasma IL-6, TNF-α and uric acid, whereas a significant reduction was found in the IL-1ß concentration and gamma-glutamyl transferase activity. Oxidative stress and antioxidant biomarkers can change throughout the season in competitive athletes, reflecting the physical stress and muscle damage that occurs as the result of competitive handball training. In addition, these biochemical measurements can be applied in the physiological follow-up of athletes.
Assuntos
Atletas , Músculo Esquelético/metabolismo , Estresse Oxidativo , Adaptação Fisiológica/fisiologia , Adulto , Animais , Desempenho Atlético , Biomarcadores/metabolismo , Exercício Físico/fisiologia , Seguimentos , Humanos , Masculino , Músculo Esquelético/lesões , Músculo Esquelético/patologia , Adulto JovemRESUMO
The present study aimed to investigate the effects of daily (45 days) intake of fish oil (FO; 10mg EPA/kg body weight (BW) and 7 mg DHA/kg BW) and/or natural ASTA (1mg ASTA/kg BW) on oxidative stress and functional indexes of neutrophils isolated from Wistar rats by monitoring superoxide (O(2)(-)), hydrogen peroxide (H(2)O(2)), and nitric oxide (NO()) production compared to the progression of auto-induced lipid peroxidation and Ca(2+) release in activated neutrophils. Furthermore, phagocytic capacity, antioxidant enzyme activities, glutathione-recycling system, and biomarkers of lipid and protein oxidation in neutrophils were compared to the redox status. Our results show evidence of the beneficial effects of FO+ASTA supplementation for immune competence based on the redox balance in plasma (significant increase in GSH-dependent reducing power), non-activated neutrophils (increased activity of the glutathione-recycling enzymes GPx and GR) and PMA-activated neutrophils (lower O(2)(-), H(2)O(2), and NO() generation, reduced membrane oxidation, but higher phagocytic activity). Combined application of ASTA and FO promoted hypolipidemic/hypocholesterolemic effects in plasma and resulted in increased phagocytic activity of activated neutrophils when compared with ASTA or FO applied alone. In PMA-activated neutrophils, ASTA was superior to FO in exerting antioxidant effects. The bulk of data reinforces the hypothesis that habitual consumption of marine fish (e.g. salmon, which is a natural source of both astaxanthin and fish oil) is beneficial to human health, in particular by improving immune response and lowering the risk of vascular and infectious diseases.
Assuntos
Óleos de Peixe/farmacologia , Glutationa/metabolismo , Neutrófilos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Animais , Clorófitas/química , Suplementos Nutricionais , Sinergismo Farmacológico , Glutationa/sangue , Peróxido de Hidrogênio/metabolismo , Masculino , Neutrófilos/citologia , Neutrófilos/metabolismo , Óxido Nítrico/metabolismo , Oxirredução/efeitos dos fármacos , Fagocitose/efeitos dos fármacos , Ratos , Ratos Wistar , Superóxidos/metabolismo , Xantofilas/farmacologiaRESUMO
PURPOSE: Higher intakes of n-3 polyunsaturated fatty acids that are abundant in marine fishes have been long described as a "good nutritional intervention" with increasing clinical benefits to cardiovascular health, inflammation, mental, and neurodegenerative diseases. The present study was designed to investigate the effect of daily fish oil (FO-10 mg EPA/kg body weight (BW) and 7 mg DHA/kg BW) intake by oral gavage associated with the antioxidant astaxanthin (ASTA-1 mg/kg BW) on the redox metabolism and the functional properties of lymphocytes from rat lymph nodes. METHODS: This study was conducted by measurements of lymphocyte proliferation capacity, ROS production [superoxide (O2(â¢-)) and hydrogen peroxide (H2O2)], nitric oxide (NO(â¢)) generation, intracellular calcium release, oxidative damage to lipids and proteins, activities of major antioxidant enzymes, GSH/GSSG content, and cytokines release. RESULTS: After 45 days of FO + ASTA supplementation, the proliferation capacity of activated T- and B-lymphocytes was significantly diminished followed by lower levels of O2(â¢-), H2O2 and NO(â¢) production, and increased activities of total/SOD, GR and GPx, and calcium release in cytosol. ASTA was able to prevent oxidative modification in cell structures through the suppression of the oxidative stress condition imposed by FO. L: -selectin was increased by FO, and IL-1ß was decreased only by ASTA supplementation. CONCLUSION: We can propose that association of ASTA with FO could be a good strategy to prevent oxidative stress induced by polyunsaturated fatty acids and also to potentiate immuno-modulatory effects of FO.
Assuntos
Antioxidantes/administração & dosagem , Suplementos Nutricionais , Óleos de Peixe/administração & dosagem , Imunomodulação , Linfócitos/imunologia , Animais , Sinalização do Cálcio/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Óleos de Peixe/química , Glutationa/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Linfócitos/metabolismo , Masculino , Mitógenos/farmacologia , Óxido Nítrico/metabolismo , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Oxirredutases/metabolismo , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Xantofilas/administração & dosagemRESUMO
Although previous studies have shown that a mixture of fatty acids in similar proportion to that found in human plasma triggers apoptosis of peripheral blood lymphocytes from healthy subjects, the mechanism involved remains unknown. In the present study, we examined whether the effect of a mixture of fatty acids upon human lymphocyte death involves cytochrome c release from the mitochondria, activation of caspases 3, 6, 8, and 9, production of superoxide anion, nitric oxide (NO), increase in cytosolic Ca(2+) levels, and expression of the anti-apoptotic 14-3-3 and the pro-apoptotic FasL, bad, and bid proteins. Peripheral blood lymphocytes from healthy subjects were isolated and treated for up to 48 h with increasing concentrations (0.1-0.4 mM) of the fatty acid mixture. Cells were then harvested and thecytochromec release from mitochondrial intermembrane space into cytosol and expression of anti- and pro-apoptotic proteins were investigated by western blot analysis. Activities of caspases 3, 6, 8, and 9 were determined using spectrofluorometric assays. NO production was monitored using DAF-2-FM probe. Cytosolic free calcium concentration ([Ca(2+)](i)) was determined using the fluorescent probe Fura-2-AM. Superoxide anion was assayed using lucigenin and dihydroethidine assays. Lymphocytes treated for 24 h with the fatty acid mixture presented increased cytochrome c release from mitochondria as compared with control lymphocytes without treatment. Activities of caspases 3, 6, and 9 were increased by 146, 22 and 35% respectively by the treatment with 0.4 mM concentration of the fatty acid mixture for 24 h. The expression of bid protein was significantly increased in lymphocytes by 40% at 0.2 mM and by 80% at 0.4 mM fatty acid concentration, whereas FasL, 14-3-3 and bad proteins were not affected by the treatment. Intracellular calcium concentration was increased in a dose-dependent manner after 30 min of fatty acid treatment and addition of BSA (0.2%) abolished this increase. Production of NO and superoxide anion was also increased by the fatty acid mixture and BSA loaded in the culture medium prevented this increase. In conclusion, fatty acids induced apoptosis of human lymphocytes by a mechanism that involved cytochrome c release from mitochondria, activation of the caspase cascade, and increase of bid protein content, superoxide and NO production, and of cytosolic calcium concentration.