Alfredo Córdova

Antioxidant response to oxidative stress induced by exhaustive exercise

The aim of this work was to demonstrate the occurrence of oxidative stress during exhaustive exercise and to determine the antioxidant response. Eight voluntary male subjects participated in this study. The exercise was a cycling mountain stage (171 km) and the cyclists took a mean+/-S.E.M. time of 270+/-12 min to complete it. Blood samples were taken before the cycling stage, immediately after the stage, 3 h after finishing the stage and on the morning of the following day. We determined the activities of erythrocyte antioxidant enzymes, blood levels of oxidised glutathione, plasma levels of antioxidant vitamins and carotenoids, and the serum lipid and cholesterol profile. The mountain cycling stage induced significant increases in catalase and glutathione reductase activities. Significant decreases in glutathione peroxidase activity, both determined with hydrogen peroxide and with cumene hydroperoxide as substrates, were observed. Blood oxidised glutathione and serum uric acid rose after the stage. Plasma vitamin E increased after the stage but dropped to below basal values after 3 h of recovery. Triglycerides and VLDL-cholesterol increased significantly after the stage and remained high 3 h after the cycling stage. The mountain cycling stage induced oxidative stress, as was evidenced by the increases in blood GSSG and in serum urate concentrations and by the pattern of change of erythrocyte antioxidant enzyme activities. A specific utilisation of alpha-tocopherol against oxidative stress during recovery was evidenced.

Relation between oxidative stress markers and antioxidant endogenous defences during exhaustive exercise

Hydrogen peroxide (H2O2) could induce oxidative damage at long distance from its generation site and it is also an important signalling molecule that induces some genes related to oxidative stress. Our objective was to study the plasma and blood cells capability to detoxify H2O2 after intense exercise and its correlation with oxidative damage. Blood samples were taken from nine professional cycling, participating in a mountain stage, under basal conditions and 3 h after the competition. Catalase and glutathione peroxidase activities decreased (40 and 50% respectively) in neutrophils after the cycling stage, while glutathione peroxidase increased (87%) in lymphocytes. Catalase protein levels and catalase specific activity maintained basal values after the stage in plasma. Catalase protein levels decreased (48%) in neutrophils and its specific activity increased up to plasma values after exercise. Myeloperoxidase (MPO) increased (39%) in neutrophils after the cycling stage. Exercise-induced hemolysis and lymphopenia inversely correlated with cellular markers of oxidative stress. Plasma malondialdehyde (MDA) directly correlated with neutrophil MPO activity and erythrocytes MDA. Intense exercise induces oxidative damage in blood cells as erythrocytes and lymphocytes, but not in neutrophils.

Sodium Nitrate Supplementation Does Not Enhance Performance of Endurance Athletes

PURPOSE Supplementation with inorganic nitrate has been suggested to be an ergogenic aid for athletes as nitric oxide donor. The purpose of this study was to determine whether ingestion of inorganic sodium nitrate benefits well-trained athletes performing a 40-min exercise test in laboratory conditions. In addition, we investigated the effect of this supplement on plasma levels of endothelin-1 (ET-1) and in nitrated proteins. METHODS Thirteen trained athletes participated in this randomized, double-blind, crossover study. They performed a 40-min cycle ergometer distance-trial test after two 3-d periods of dietary supplementation with sodium nitrate (10 mg·kg of body mass) or placebo. RESULTS Concentration of plasma nitrate (256 ± 35 μM) and nitrite (334 ± 86 nM) increased significantly (P < 0.05) after nitrate supplementation compared with placebo (nitrate: 44 ± 11 μM; nitrite: 187 ± 43 nM). In terms of exercise performance, there were no differences in either the mean distance (nitrate: 26.4 ± 1.1 km; placebo: 26.3 ± 1.2 km; P = 0.61) or mean power output (nitrate: 258 ± 28 W; placebo: 257 ± 28 W; P = 0.89) between treatments. Plasma ET-1 increased significantly (P < 0.05) just after exercise in nitrate (4.0 ± 0.8 pg·mL) and placebo (2.4 ± 0.4 pg·mL) conditions. This increase was significantly greater (P < 0.05) in the nitrate group. Levels of nitrated proteins did not differ between treatments (nitrate: preexercise, 91% ± 23%; postexercise, 81% ± 23%; placebo: preexercise, 95% ± 20%; postexercise, 99% ± 19%). CONCLUSION Sodium nitrate supplementation did not improve a 40-min distance-trial performance in endurance athletes. In addition, concentration of plasma ET-1 increased significantly after exercise after supplementation with sodium nitrate.

Effect of exercise intensity and training on antioxidants and cholesterol profile in cyclists.

The aim of this work was to evaluate the effect of different intensity of exercise and different training status on antioxidants and cholesterol profile in cyclists. 33 male cyclists (17 amateur and 16 professional cyclists) participated in this study. The amateurs all trained 14 +/- 1 h each week, and their VO(2) max was 62.5 +/- 1.8 ml/Kg x min; the professionals all trained 24 +/- 1 h each week, and their VO(2) max was 80.2 +/- 1.6 ml/Kg x min. Amateurs were submitted to the maximal and submaximal prolonged exercise tests. Professionals were submitted to a mountain stage (170 km) of cycling competition. Serum lipid and cholesterol profile (triglycerides, total cholesterol, HDL-cholesterol, LDL-cholesterol and VLDL-cholesterol) and plasma antioxidant capacity (ascorbic acid, alpha-tocopherol, retinol, beta-carotene and others) were measured before and after exercise tests. Hematological determinations (number of erythrocytes, hematocrit and hemoglobin concentration) and dietary intake were also measured. No significant differences were observed in basal values (before exercise tests) of amateur and professional cyclists. Negligible differences were found between dietary intake of amateur and professional cyclists, and also the results of hematological values showed there was no effect of degree of hydration or dietary intake on blood levels of studied antioxidant and lipid parameters. An increase in plasma levels of vitamin C, vitamin E, triglycerides and VLDL-cholesterol levels, and also a decrease of beta-carotene and LDL-cholesterol. were observed in well-trained professional cyclists after the cycling stage - an endurance exercise--but not in amateur cyclists. Amateur cyclists showed only mild increases in total cholesterol after maximal and submaximal exercise, while a rise in HDL-cholesterol was only observed after maximal exercise; none of these changes were observed in professional cyclists. Plasma levels of antioxidant vitamins and carotenes, and also serum lipids, total cholesterol and lipoprotein-cholesterol showed an overall response to exercise, and their increase and/or decrease must be explained as a consequence of the different training status of sportsmen and intensity and duration of exercise tests.

Response of antioxidant defences to oxidative stress induced by prolonged exercise: antioxidant enzyme gene expression in lymphocytes

The response of lymphocyte and plasma antioxidant defences to a prolonged exercise as a cycling stage in a professional race was analysed. Antioxidant enzyme activities and gene expression, carbonyl derivative and MDA levels were determined in lymphocytes. Plasma levels of vitamin E, carotenes, protein carbonyl derivatives and the test d-Roms were measured. Significant increases in plasmatic carbonyls and in the test d-Roms were observed after the cycling stage. No significant differences were found in the lymphocyte MDA and carbonyl derivative levels. A significant increase was found in plasma vitamin E concentration after the cycling stage; however, the lymphocyte vitamin E concentration did not change. Significant increases were observed in lymphocyte total superoxide dismutase (SOD) activity and in the levels of CuZn-SOD and Mn-SOD isoenzymes. The moderate levels of oxidative stress in the lymphocyte induced a cellular adaptation to exercise enough to counteract the negative effects of oxidative stress.

Effect of Training on Zinc Metabolism: Changes in Serum and Sweat Zinc Concentrations in Sportsmen

The purpose of this research was to determine the effects of daily physical training on serum and sweat zinc concentrations in professional sportsmen between October and December, during the competing season. Twelve volleyball players and another 12 control subjects have participated in this study. Tests were made in October and December which consisted of a progressive bicycle ergometer test (increasing 30 W every 3 min to reach maximum tolerated power). Blood samples were obtained at rest and immediately after exercise. Total serum zinc increased significantly after maximal exercise in both sportsmen and control subjects. In athletes, the change after exercise was significantly higher in December than in October. The percentage of ultrafiltrable zinc (ZnUf) in October was similar in sportsmen and in controls. In December, however, after exercise, the percentage of ZnUf was higher in athletes. With respect to sweat zinc, it was in the same range both in controls and in sportsmen in October. In December, however, sweat zinc was significantly higher in athletes as compared with the situation in October and with respect to the control group. In October, the zinc concentration of urine was similar for sportsmen and controls. In December, the sportsmen showed an increase in urinary zinc excretion with respect to control subjects. Cortisol in athletes increased significantly after exercise in December. In conclusion, a daily and maintained practice of exercise is probably responsible for an alteration of zinc metabolism. The results suggest that ZnUf control, zinc supplementation and/or stress control appear to be indicated in athletes to prevent the diminution of active ZnUf. In our practical opinion we think that alterations in zinc metabolism with increases in zinc excretion and stress levels lead to a situation of latent fatigue with a decreased endurance.

Acute phase immune response to exercise coexists with decreased neutrophil antioxidant enzyme defences

Long-duration or damaging exercise initiates reactions that resemble the acute phase response to infection and induces neutrophil priming for oxidative activity. Our objective was to establish the status of the antioxidant defences and of the oxidative equilibrium in the neutrophils of sportsmen prior to and after intense physical exercise. Nine voluntary male professional cyclists participated in this study. The exercise was a cycling mountain stage (171 km) and the cyclists took a mean - SEM of 270 - 12 min to complete it. We determined the activities of catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), the levels and activity of superoxide dismutase (SOD), the concentrations of ascorbate, glutathione and glutathione disulphide (GSSG) and DNA levels in neutrophils. The cycling stage decreased enzyme activities expressed per DNA units: CAT (33%), SOD (38%), GPx (65%); increased ascorbate concentration in neutrophils and decreased the GSH/GSSG ratio and the enzyme activities expressed per DNA units. Neutrophils could contribute to plasma antioxidant defences against oxidative stress induced by exercise because they probably provide antioxidant enzymes and ascorbate.

Effects of L-citrulline oral supplementation on polymorphonuclear neutrophils oxidative burst and nitric oxide production after exercise

Seventeen volunteer male professional cyclists were randomly assigned to control or supplemented (6 g L-citrulline-malate) groups and participated in a cycling stage. Blood samples were taken in basal conditions, after the race and 3 h post-race. Citrulline supplementation significantly increased plasma concentration of both arginine and citrulline after the stage only in the supplemented group. Polymorphonuclear neutrophils (PMNs) from controls responded to exercise with a progressive decrease in ROS production. Supplemented PMNs significantly increased ROS production after exercise compared to basal values and diminished to values lower than basal at recovery. PMN nitrite concentration was significantly higher after exercise and recovery only in the supplemented group. Markers of oxidative damage—CK, LDH, malondialdehyde—and DNA damage remained unchanged in both groups. In conclusion, oral L-citrulline administration previous to a cycling stage increases plasma arginine availability for NO synthesis and PMNs priming for oxidative burst without oxidative damage.

Pre-exercise antioxidant enzyme activities determine the antioxidant enzyme erythrocyte response to exercise

Free radical production increases during exercise and oxidative damage occurs in several tissues. We examined the effects of three different exercise tests on the pattern of change of erythrocyte enzyme antioxidant activities. The tests were a short maximal exercise test, a submaximal prolonged exercise test and a cycling stage during competition. The participants were amateur and professional cyclists with different training statuses and different basal erythrocyte antioxidant enzyme activities. The maximal test produced no changes in the erythrocyte antioxidant enzyme activities of amateur sportsmen. The submaximal test, performed at 80% of maximal oxygen uptake, decreased erythrocyte catalase (12%), glutathione peroxidase determined with H2O2 (14%) and glutathione reductase (16%); superoxide dismutase activity increased by about 25%. The cycling stage performed by professional cyclists increased erythrocyte catalase (29%) and glutathione reductase (10%) activities. The in vivo changes in glutathione reductase activity were confirmed by in vitro measurements: hydrogen peroxide decreased and the presence of catalase increased the activity of this enzyme. In conclusion, we suggest that the different erythrocyte antioxidant enzyme responses to diverse exercise tests can be explained by the effects of hydrogen peroxide and the superoxide anion on the antioxidant enzyme activities in erythrocytes.

Intense physical activity enhances neutrophil antioxidant enzyme gene expression. Immunocytochemistry evidence for catalase secretion

We studied the effects of intense exercise on the neutrophil antioxidant enzyme activities and gene expression. Blood samples were taken from seven cyclists in basal conditions and 3 h after two competition stages of 165 km. Serum creatine kinase (CK) activity, plasma carbonyl derivatives and uric acid levels increased after exercise. The cycling stage induced neutrophilia and increased myeloperoxidase (MPO) activity and reactive oxygen species (ROS) production. Antioxidant enzyme activities (catalase, glutathione peroxidase and superoxide dismutase) decreased after exercise, although gene expression increased. Immunocytochemistry showed catalase (CAT) enzyme equally distributed between the cytoplasm and organelles before exercise, and after exercise the cytoplasmic CAT levels were reduced and were absent in the compartments. After in vitro stimulation with opsonized zymosan (OZ) the extracellular CAT levels increased. This suggests a CAT secretion in order to avoid neutrophil-induced oxidative damage at a local level or to regulate the function of ROS as extracellular signalling molecules.