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Abstract
The objective of the present study was to evaluate a comprehensive set of urinary biomarkers for oxidative damage to lipids, proteins and DNA, in man. Eighteen moderately trained males (mean age 24.6±0.7) exercised 60 min at 70% of maximal O2 uptake on a cycle ergometer. Urine fractions for 12 h were collected 1 day before, and for 3 consecutive days after exercise.
As biomarkers of lipid peroxidation, 8 aldehydes (i.e. propanal, butanal, pentanal, hexanal, heptanal, octanal, nonanal and malondialdehyde—MDA)and acetone were analyzed in urines by gas chromatography with electron capture detection (GC-ECD). As a biomarker of protein oxidation, o,o′-dityrosine was analyzed in urine samples by a recently developed isotope dilution HPLC-atmospheric pressure chemical ionization (APCI)-tandem-mass spectrometry (HPLC-APCI-MS/MS) methodology. As a biomarker of oxidative DNA damage, urinary excretion of 8-hydroxy-2′-deoxyguanosine (8-OHdG) was measured by an ELISA method.
On the day of exercise, significant increases were observed in urinary excretions of acetone ( p<0.025, n=18) and butanal ( p<0.01, n=18) in the 12 h daytime fractions compared to the daytime fraction before exercise. The urinary acetone excretion was also significantly ( p<0.05) increased on the 1st day after exercise. Octanal and nonanal were increased in the daytime urine fraction on the 2nd day after exercise. However, these increases were of borderline significance ( p=0.09 and p=0.07, respectively).
Significantly elevated urinary o,o′-dityrosine amounts were observed in the daytime fraction on the day of exercise ( p<0.025) and on the 1st day after exercise ( p=0.07) compared to the before exercise daytime fraction.
Excretion of urinary 8-OHdG was statistically significantly increased in the daytime fractions on the day of exercise ( p=0.07) and on the 1st day after exercise ( p<0.025) compared to before exercise daytime fraction.
Increases in urinary excretions of acetone, propanal, pentanal, MDA and 8-OHdG significantly correlated with training status (hours of exercise/week) of the volunteers, while o,o′-dityrosine did not.
To our knowledge, the present study is the first to evaluate a multi-parameter non-invasive biomarker set for damage to three main cellular targets of ROS. It shows that 1 h of exercise may already induce oxidative damage in moderately trained individuals and that the chosen urinary biomarkers are sensitive enough to monitor such damage.
