Original Contribution
Vitamin C supplementation lowers urinary levels of 4-hydroperoxy-2-nonenal metabolites in humans

https://doi.org/10.1016/j.freeradbiomed.2011.01.004 Get rights and content

Abstract

The lack of suitable biomarkers of oxidative stress is a common problem for antioxidant intervention studies in humans. We evaluated the efficacy of vitamin C supplementation in decreasing biomarkers of lipid peroxidation in nonsmokers and in cigarette smokers, a commonly studied, free-living human model of chronic oxidative stress. Participants received ascorbic acid (500 mg twice per day) or placebo for 17 days in a double-blind, placebo-controlled, randomized crossover design study. The urinary biomarkers assessed and reported herein are derived from 4-hydroperoxy-2-nonenal (HPNE) and include the mercapturic acid (MA) conjugates of 4-hydroxy-2(E)-nonenal (HNE), 1,4-dihydroxy-2(E)-nonene (DHN), and 4-oxo-2(E)-nonenol(ONO). Vitamin C supplementation decreased the urinary concentrations of both ONO-MA (p = 0.0013) and HNE-MA (p = 0.0213) by ~ 30%; however, neither cigarette smoking nor sex affected these biomarkers. In contrast, vitamin C supplementation decreased urinary concentrations of DHN-MA (three-way interaction p = 0.0304) in nonsmoking men compared with nonsmoking women (p < 0.05), as well as in nonsmoking men compared with smoking men (p < 0.05). Vitamin C supplementation also decreased (p = 0.0092) urinary total of metabolites by ~ 20%. Thus, HPNE metabolites can be reduced favorably in response to improved plasma ascorbic acid concentrations, an effect due to ascorbic acid antioxidant function.

Section snippets

Materials

HNE and ONO as well as their mercapturic acid (MA) and N-(acetyl-d3)-L-cysteine (MAd3) conjugates were prepared and chemically characterized as described previously [18], [25]. DHN-MA and DHN-MAd3 were prepared by NaBH4 reduction of HNE-MA and HNE-MAd3 [18]. HPLC-grade solvents were obtained from Honeywell Burdick and Jackson (Muskegon, MI, USA) (0.1% formic acid in water) and from EMD Chemicals (San Diego, CA, USA) (acetonitrile). All other chemicals were obtained from Sigma–Aldrich (St.

Participant characteristics and plasma vitamin C

Urinary HPNE metabolites were measured in samples collected throughout a study examining vitamin C supplementation on vitamin E pharmacokinetics. Complete details of this study have been published elsewhere [26]. In brief, vitamin C supplementation ameliorated the faster rate of vitamin E disappearance in smokers, but had no effect on nonsmokers’ vitamin E kinetics [26]. No significant differences (p > 0.05) were observed between nonsmokers and smokers with respect to age or body mass index (BMI)

Discussion

The findings of this study demonstrate for the first time that vitamin C supplementation dramatically reduces urinary concentrations of HPNE metabolites in a cohort of healthy nonsmokers and smokers. Urinary levels of ONO-MA and HNE-MA, independent of sex or smoking status, are reduced by ~ 30% after vitamin C supplementation (Fig. 4). Similarly, vitamin C supplementation decreased the total of HPNE metabolites by ~ 20%. These findings can be explained by the antioxidant effects of vitamin C,

Acknowledgments

This study was supported by NIH Grants R01HL081721 (J.F.S.), R01DK059576 (M.G.T.), R01DK067930 (M.G.T.), and S10RR022589 (J.F.S.). The authors acknowledge the use of the Mass Spectrometry Facility of the Environmental Health Sciences Center at Oregon State University (NIH Grant P30ES000210).

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  • Cited by (0)

    1

    Present address: Center for Disease Control and Prevention, Atlanta, GA, USA.

    2

    Present address: Department of Nutritional Sciences, University of Connecticut, Storrs, CT, USA.

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