Douglas Popp Marin

ROS production in neutrophils from alloxan-induced diabetic rats treated in vivo with astaxanthin

BACKGROUND Astaxanthin (ASTA) is a carotenoid which has powerful antioxidant, anti-tumor, anti-diabetic, anti-inflammatory and cardioprotective properties. The present study investigated the effect of daily ASTA intake on oxidative stress and the functional properties of neutrophils from alloxan-induced diabetic rats. METHODS Neutrophils isolated from ASTA-fed rats (30days, 20mg ASTA/kg of body weight - BW) induced to diabetes by alloxan treatment (i.p. 75mg/BW) were assessed by: production of superoxide and hydrogen peroxide, nitric oxide, basal calcium release, oxidative damage (TBARS and carbonyls content), and activities of major antioxidant enzymes. RESULTS Our results show that diabetes promotes a significant oxidative stress in neutrophils. The production of superoxide was significantly increased in neutrophils from diabetic rats and treatment with ASTA was not effective in reducing superoxide levels. At the same time, a reduction in the activity of total superoxide dismutase enzyme was observed, which was not restored after treatment with ASTA. At resting conditions, neutrophils have a higher basal production of hydrogen peroxide, which is enhanced following PMA-stimulation. Treatment with ASTA does not restore values to the basal levels. The indicators of oxidative damage to biomolecules showed that diabetic rats significantly increased the lipid and protein damage, but this change was reversed after treatment with ASTA. CONCLUSION Our results show that diabetes condition promotes a marked oxidative stress in neutrophils and treatment with ASTA for 30days at a dose of 20mg/kg of BW partially reverses those deleterious effects.

Cytokines and Oxidative Stress Status Following a Handball Game in Elite Male Players

Background. Handball is considered an intermittent sport that places an important stress on a players aerobic and anaerobic metabolism. However, the oxidative stress responses following a handball game remain unknown. We investigated the responses of plasma and erythrocyte antioxidant system and oxidative stress biomarkers following a single handball game. Methods. Fourteen male elite Brazilian handball athletes were recruited in the present study. Blood samples were taken before, immediately, and 24 hours after the game. Results. After the game and during 24 hours of recovery, the concentration of all oxidative stress indices changed significantly in a way indicating increased oxidative stress in the blood (thiol groups and reduced glutathione decreased, whereas TBARS and plasma antioxidant capacity was increased) as well as in erythrocyte (increased levels of TBARS and protein carbonyls). Erythrocyte antioxidant enzyme activities were also significantly changed by handball. Muscle damage indices (creatine kinase and lactate dehydrogenase) increased significantly after exercise. In addition, IL-6 increased after the game, whereas TNF-α decreased during recovery. Conclusion. This study demonstrates that a single handball game in elite athletes induces a marked state of oxidative stress evidenced by the oxidative modification in plasma and erythrocyte macromolecules, as well as by changes in the enzymatic and nonenzymatic antioxidant system.

Testosterone suppresses oxidative stress in human neutrophils

The in vitro effect of testosterone on human neutrophil function was investigated. Blood neutrophils from healthy male subjects were isolated and treated with 10 nM, 0.1 and 10 µM testosterone for 24 h. As compared with untreated cells, the testosterone treatment produced a significant decrease of superoxide production as indicated by the measurement of extra‐ and intracellular superoxide content. An increment in the production of nitric oxide was observed at 0.1 and 10 µM testosterone concentrations, whereas no effect was found for 10 nM. Intracellular calcium mobilization was significantly increased at 10 nM, whereas it was reduced at 10 µM testosterone. There was an increase in phagocytic capacity at 10 nM and a decrease of microbicidal activity in neutrophils treated with testosterone at 10 µM. Glutathione reductase activity was increased by testosterone treatment, whereas no effect was observed in other antioxidant enzyme activities. An increase in the content of thiol groups was observed at all testosterone concentrations. Lipid peroxidation in neutrophils evaluated by levels of TBARS was decreased at 10 nM and 0.1 µM testosterone. These results indicate the antioxidant properties of testosterone in neutrophils as suggested by reduction of superoxide anion production, and lipid peroxidation, and by the increase in nitric oxide production, glutathione reductase activity and the content of thiol groups. Therefore, the plasma levels of testosterone are important regulators of neutrophil function and so of the inflammatory response. Copyright

Oxidative stress and antioxidant status response of handball athletes: Implications for sport training monitoring☆

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-1β 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.

Combined astaxanthin and fish oil supplementation improves glutathione-based redox balance in rat plasma and neutrophils

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.

Effects of chronic resveratrol supplementation in military firefighters undergo a physical fitness test – A placebo-controlled, double blind study

PURPOSE The purpose of this study was to determine the plasma metabolic response and certain indicators of oxidative stress (antioxidant system and oxidative stress biomarkers) in plasma and erythrocytes of Brazilian military firefighters supplemented or not with resveratrol (RES) for 90 days (100 mg/day). The analyses were performed before and after a typical physical fitness test (FT) used to induce oxidative stress. METHODS/RESULTS In this placebo-controlled double-blinded study, we observed that RES supplementation did not present hepatic consequences compared with the placebo group following analysis of AST, ALT and GGT plasma activities. Plasma glucose and triglycerides levels were increased after the FT in firefighters supplemented with RES but were not elevated at baseline. Neither total nor cholesterol fractions were modified by RES supplementation. CK levels were increased after the firefighters performed the FT; however, no differences were determined between the placebo and RES groups. Ferric-reducing ability of plasma as well as uric acid was increased after the FT, but was not modified by RES supplementation. Plasma oxidative stress biomarkers, such as thiol content, 8-isoprostane and 8OHdG, showed no modifications, while IL-6 and TNF-α were decreased in the RES group after the FT. Among antioxidant enzyme activities determined in erythrocytes from the firefighters, only GPx activity was reduced by RES supplementation both before and after the FT. CONCLUSION In summary, the most pronounced effect of RES supplementation is its anti-inflammatory effect, which reduced IL-6 and TNF-α level. The FT applied to Brazilian military firefighters was not sufficient to challenge the antioxidant defense systems, and, therefore, 100mg of RES for three months did not induce significant effects.

Phenanthrene decreases neutrophil function by disrupting intracellular redox balance

The aim of the present work was to evaluate whether the treatment of human neutrophils with phenanthrene (PHN) can alter the phagocytic and microbicidal capacity of these cells by causing a disruption in redox balance. Peripheral neutrophils from healthy subjects were treated for up to 24 h with increasing concentrations of phenanthrene. Phagocytic/microbicidal activities, antioxidant enzymes, oxidative lesions (thiobarbituric acid‐reactive substances and protein thiol and carbonyl groups) and redox signaling compounds (intracellular Ca2+, superoxide, hydrogen peroxide and nitric oxide) were monitored on neutrophils exposed to 10 µg PHN ml−1. Cell viability decreased abruptly at PHN concentrations above 10 µg ml−1 (LC50 = 20.86 ± 0.51 µg ml−1 and p‐sigmoidal slope = 19.88 ± 10.11). Phagocytic and microbicidal capacities were decreased by 60 and 82%, respectively. Substantial increases in total‐/Mn‐SOD, catalase, glutathione peroxidase and glutathione reductase activities (by 61, 15, 87, 245 and 70%, respectively) matched the oxidative injury obtained in TBARS (2.5‐fold higher) and protein thiol (54% lower). Diminished productions of superoxide by 18% and hydrogen peroxide by 29% were observed in association to exacerbated calcium (27%) and nitric oxide (63%) levels. The data indicate that phenanthrene might be associated with substantial reduction in human neutrophil functions due to severe intracellular redox imbalances. Copyright