ABSTRACT
Several studies suggest that exercise is associated with elevated oxidative stress which diminishes NO bioavailability. The aim of the present study was to investigate a potential link between NO synthesis and bioavailability and oxidative stress in the circulation of subjects performing high-intensive endurance exercise. Twenty-two male healthy subjects cycled at 80% of their maximal workload. Cubital venous blood was taken before, during and after exercise, and heparinized plasma was generated. Plasma concentrations of nitrite and nitrate were quantified by GC-MS and of the oxidative stress biomarker 15(S)-8-iso-PGF(2alpha) by GC-MS/MS. pH and pCO(2) fell and HbO(2) increased upon exercise. The duration of the 80% phase (d80) was 740+/-210s. Subjects cycled at 89.2+/-3.3% of their peak oxygen uptake. Plasma concentration of nitrite (P<0.01) and 15(S)-8-iso-PGF(2alpha) (P<0.05) decreased significantly during exercise. At the end of exercise, plasma nitrite concentration correlated positively with d80 and performed work (w80) (each P<0.05). Changes in nitrate concentration also correlated positively with d80 (P<0.05) and w80/kg (P<0.01). These findings provide evidence of a favorable effect of nitrite on high-intensive endurance exercise. The lack of association between 15(S)-8-iso-PGF(2alpha) and NO bioavailability (nitrite concentration) and NO biosynthesis (nitrate concentration) suggest that oxidative stress, notably lipid peroxidation, is not linked to the l-arginine/NO pathway in healthy male subjects being on endurance exercise.
Subject(s)
Exercise/physiology , Health , Nitrates/blood , Nitrites/blood , Oxidative Stress/physiology , Physical Endurance/physiology , Adult , Biomarkers/blood , Carbon Dioxide/blood , Carbon Dioxide/metabolism , Dinoprost/analogs & derivatives , Dinoprost/blood , Dinoprost/metabolism , Exercise Test , Feasibility Studies , Gas Chromatography-Mass Spectrometry , Hemoglobins/metabolism , Humans , Hydrogen-Ion Concentration , Lipid Peroxidation/physiology , Male , Nitrates/metabolism , Nitric Oxide/biosynthesis , Nitric Oxide/metabolism , Nitrites/metabolism , Oxygen/metabolism , Reference Values , Time FactorsABSTRACT
OBJECTIVES: Malondialdehyde (MDA) and the F(2)-isoprostane 15(S)-8-iso-prostaglandin F(2alpha) (15(S)-8-iso-PGF(2alpha)) belong to the most frequently analyzed biomarkers of oxidative stress in basic and clinical research. The objective of the present study was to examine the effect of hemolysis on free MDA and total (free+esterified) 15(S)-8-iso-PGF(2alpha) concentrations in human plasma. DESIGN AND METHODS: MDA and 15(S)-8-iso-PGF(2alpha) were determined by GC-MS/MS in plasma samples from venous heparinized blood drawn under resting conditions (n=22) as well as under physical exercise (n=158) in 22 healthy young subjects. In vitro, we prepared plasma samples with hemolysis degrees up to 0.8% using artificially hemolyzed, freshly obtained heparinized blood. RESULTS: In some plasma samples of the exercise study both under resting and exercise conditions, clinically significant hemolysis was macroscopically visible. Both in vivo (r=0.74) and in vitro (r=0.87), we found a significant positive correlation between hemolysis degree (0-0.2%) and MDA plasma concentrations (50-250 nmol/L). Unlike in vitro (r=0.84), in vivo, 15(S)-8-iso-PGF(2alpha) and MDA plasma concentrations correlated weakly (r=0.50). CONCLUSIONS: We hypothesize that free hemoglobin catalyzes the formation of MDA and 15(S)-8-iso-PGF(2alpha) from free and esterified arachidonic acid. Plasma concentrations of MDA and total 15(S)-8-iso-PGF(2alpha) may be markedly compromised by hemolysis. Measurements of MDA and 15(S)-8-iso-PGF(2alpha) should be treated with caution regarding involvement of oxidative stress in disease as well as in health both under resting conditions and under exercise.
