Advertisement

European Journal of Nutrition

, Volume 44, Issue 7, pp 422–428 | Cite as

Interactive effects of polyphenols, tocopherol and ascorbic acid on the Cu2+–mediated oxidative modification of human low density lipoproteins

  • V. C. Yeomans
  • J. Linseisen
  • G. Wolfram
ORIGINAL CONTRIBUTION

Summary

Background

Only limited knowledge is available about any interactions between phenolic compounds and other antioxidants in inhibiting LDL oxidation. Many foods and beverages contain high levels of phenolic compounds; therefore, these compounds should not be considered in isolation from each other.

Aim of the study

The aim of this study was to examine the structure–antioxidant activity relationship of quercetin, caffeic acid, epicatechin, hesperetin and phloretin as well as α–tocopherol and ascorbic acid through their ability to interact with copper ions.

Methods

Isolated human LDL were incubated with single antioxidants or a combination of two and the kinetics of lipid peroxidation were assessed by measurement of conjugated diene formation (lag phase) via monitoring the absorbance at 234 nm after addition of copper ions. In addition, the degree of oxidation of the LDL protein moiety was followed by tryptophan fluorescence and carbonyl content measurements.

Results

α-Tocopherol and ascorbic acid showed a lower antioxidant activity in all test systems as compared to polyphenols at equimolar concentrations. Quercetin was the most effective compound in all three systems (p < 0.001 for lag phase and carbonyl content determination). A significant (p < 0.001) prolongation of the lag phase was found when combinations of ascorbic acid/quercetin, ascorbic acid/epicatechin, epicatechin/caffeic acid, and quercetin/epicatechin were tested as compared to the sum of the individual effects. Concerning the effects on LDL protein oxidation, the results from carbonyl content and the tryptophan fluorescence measurements showed that the combination of quercetin and caffeic acid revealed the strongest inhibitory effect (p < 0.001 carbonyl content; p ≤ 0.002 tryptohan fluorescence) on protein oxidation which was higher than the effect of the single compounds.

Conclusions

The results of the present study indicate that a combination of different antioxidants can be superior to the action of single antioxidants in protecting LDL lipid and protein moiety against oxidation. However, the substances may act by different antioxidative mechanisms, which are not necessarily complementary.

Key words

low density lipoproteins oxidation lipids protein polyphenols antioxidants tocopherol ascorbic acid 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Morton LW, bu-Amsha CR, Puddey IB, Croft KD (2000) Chemistry and biological effects of dietary phenolic compounds: relevance to cardiovascular disease. Clin Exp Pharmacol Physiol 27:152–159CrossRefPubMedGoogle Scholar
  2. 2.
    Noguchi N, Niki E (2000) Phenolic antioxidants: a rationale for design and evaluation of novel antioxidant drug for atherosclerosis. Free Radic Biol Med 28:1538–1546CrossRefPubMedGoogle Scholar
  3. 3.
    Halliwell B (1994) Free radicals, antioxidants, and human disease: curiosity, cause, or consequence? Lancet 344:721–724CrossRefPubMedGoogle Scholar
  4. 4.
    Pinchuk I, Gal S, Lichtenberg D (2001) The dose-dependent effect of copperchelating agents on the kinetics of peroxidation of low-density lipoprotein (LDL). Free Radic Res 34:349–362PubMedGoogle Scholar
  5. 5.
    Esterbauer H, Ramos P (1996) Chemistry and pathophysiology of oxidation of LDL. Rev Physiol Biochem Pharmacol 127:31–64PubMedGoogle Scholar
  6. 6.
    Halliwell B, Gutteridge JM (1990) The antioxidants of human extracellular fluids. Arch Biochem Biophys 280:1–8CrossRefPubMedGoogle Scholar
  7. 7.
    Croft KD (1998) The chemistry and biological effects of flavonoids and phenolic acids. Ann N Y Acad Sci 20(854):435–442Google Scholar
  8. 8.
    Evans PJ, Smith C, Mitchinson MJ, Halliwell B (1995) Metal ion release from mechanically-disrupted human arterial wall. Implications for the development of atherosclerosis. Free Radic Res 23:465–469PubMedGoogle Scholar
  9. 9.
    Smith C, Mitchinson MJ, Aruoma OI, Halliwell B (1992) Stimulation of lipid peroxidation and hydroxyl-radical generation by the contents of human atherosclerotic lesions. Biochem J 286:901–905PubMedGoogle Scholar
  10. 10.
    Halliwell B, Aeschbach R, Loliger J, Aruoma OI (1995) The characterization of antioxidants. Food Chem Toxicol 33:601–617CrossRefPubMedGoogle Scholar
  11. 11.
    Havel RJ, Eder HA, Bragdon JH (1955) The distribution and chemical composition of ultracentrifugally seperated lipoproteins in human serum. J Clin Invest 34:1345–1353PubMedGoogle Scholar
  12. 12.
    Lowry OH, Rosebrough NJ, Farr L, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  13. 13.
    Hess D, Keller HE, Oberlin B, Bonfanti R, Schuep W (1991) Simultaneous determination of retinol, tocopherols, carotenes and lycopene in plasma by means of high-performance liquid chromatography on reversed phase. Int J Vitam Nutr Res 61:232–238PubMedGoogle Scholar
  14. 14.
    Esterbauer H, Striegl G, Puhl H, Rotheneder M (1989) Continuous monitoring of in vitro oxidation of human low density lipoprotein. Free Radic Res Commun 6:67–75PubMedGoogle Scholar
  15. 15.
    Yan LJ, Traber MG, Packer L (1995) Spectrophotometric method for determination of carbonyls in oxidatively modified apolipoprotein B of human low-density lipoproteins. Anal Biochem 228:349–351CrossRefPubMedGoogle Scholar
  16. 16.
    Reznick AZ, Packer L (1994) Oxidative damage to proteins: spectrophotometric method for carbonyl assay. Methods Enzymol 233:357–363PubMedGoogle Scholar
  17. 17.
    Yan LJ, Lodge JK, Traber MG, Packer L (1997) Apolipoprotein B carbonyl formation is enhanced by lipid peroxidation during copper-mediated oxidation of human low-density lipoproteins. Arch Biochem Biophys 339:165–171CrossRefPubMedGoogle Scholar
  18. 18.
    Horakova L, Giessauf A, Raber G, Esterbauer H (1996) Effect of stobadine on Cu(++)-mediated oxidation of lowdensity lipoprotein. Biochem Pharmacol 51:1277–1282CrossRefPubMedGoogle Scholar
  19. 19.
    Giessauf A, Steiner E, Esterbauer H (1995) Early destruction of tryptophan residues of apolipoprotein B is a vitamin E-independent process during copper-mediated oxidation of LDL. Biochim Biophys Acta 1256:221–232PubMedGoogle Scholar
  20. 20.
    Cao G, Sofic E, Prior RL (1996) Antioxidant capacity of tea and common vegetables. J Agric Food Chem 44:3426–3431CrossRefGoogle Scholar
  21. 21.
    Wang H, Cao G, Prior RL (1996) Total antioxidant capacity of fruits. J Agric Food Chem 44:701–705Google Scholar
  22. 22.
    Manach C, Morand C, Crespy V, Demigne C, Texier O, Regerat F, Rémésy C (1998) Quercetin is recovered in human plasma as conjugated derivatives which retain antioxidant properties. FEBS Lett 426:331–336CrossRefPubMedGoogle Scholar
  23. 23.
    Radtke J, Linseisen J, Wolfram G (2002) Fasting plasma concentrations of selected flavonoids as markers of their ordinary dietary intake. Eur J Nutr 41(5):203–209CrossRefPubMedGoogle Scholar
  24. 24.
    Erlund I, Meririnne E, Alfthan G, Aro A (2001) Plasma kinetics and urinary excretion of the flavanones naringenin and Hesperetin in humans after ingestion of orange juice and grapefruit juice. J Nutr 131:235–241PubMedGoogle Scholar
  25. 25.
    Rein D, Lotito S, Holt R, Keen CL, Schmitz HH, Fraga CG (2000) Epicatechin in human plasma: in vivo determination and effect of chocolate consumption on plasma oxidation status. J Nutr 130:2109S–2114SPubMedGoogle Scholar
  26. 26.
    Lee MJ, Maliakal P, Chen L, Meng X, Bondoc FY, Prabhu S, Lambert G, Mohr S, Yang CS (2002) Pharmacokinetics of tea catechins after ingestion of green tea and (-)-epigallocatechin-3-gallate by humans: formation of different metabolites and individual variability. Cancer Epidemiol Biomarkers Prev 11(10 Pt 1):1025–1032PubMedGoogle Scholar
  27. 27.
    Paganga G, Rice-Evans CA (1997) The identification of flavonoids as glycosides in human plasma. FEBS Lett 13; 401(1):78–82CrossRefGoogle Scholar
  28. 28.
    Böhm H, Boeing H, Hempel J, Raab B, Kroke A (1998) Flavonols, flavone and anthocyanins as natural antioxidants of food and their possible role in the prevention of chronic diseases. Z Ernährungswiss 37:147–163PubMedGoogle Scholar
  29. 29.
    Laranjinha J, Cadenas E (1999) Redox cycles of caffeic acid, alpha-tocopherol, and ascorbate: implications for protection of low-density lipoproteins against oxidation. IUBMB Life 48:57–65PubMedGoogle Scholar
  30. 30.
    Sekher PA, Chan TS, O’Brien PJ, Rice- Evans CA (2001) Flavonoid b-ring chemistry and antioxidant activity: fast reaction kinetics. Biochem Biophys Res Commun 282:1161–1168PubMedGoogle Scholar
  31. 31.
    van Acker SA, Bast A, van der Vijgh WJF (1998) Structural aspects of antioxidant activity of flavonoids. In: Rice-Evans CA, Packer L (eds) Flavonoids in health and disease. Marcel Dekker Inc., New York, pp 221–252Google Scholar
  32. 32.
    Rice-Evans CA, Miller NJ, Paganga G (1996) Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Radic Biol Med 20:933–956CrossRefPubMedGoogle Scholar
  33. 33.
    Pietta PG (2000) Flavonoids as antioxidants. J Nat Prod 63:1035–1042CrossRefPubMedGoogle Scholar
  34. 34.
    Rice-Evans C, Miller NJ (1998) Structure- Antioxidant Activity Relationships of Flavonoids and Isoflavonoids. In: Rice-Evans C, Packer L (eds) Flavonoids in Health and Disease. Marcel Dekker, Inc., New York, pp 199–219Google Scholar
  35. 35.
    Bors W, Michel C, Schikora S (1995) Interaction of flavonoids with ascorbate and determination of their univalent redox potentials: a pulse radiolysis study. Free Radic Biol Med 19:45–52CrossRefPubMedGoogle Scholar
  36. 36.
    Cossins E, Lee R, Packer L (1998) ESR studies of vitamin C regeneration, order of reactivity of natural source phytochemical preparations. Biochem Mol Biol Int 45:583–597PubMedGoogle Scholar
  37. 37.
    Jovanovic SV, Steenken S, Simic MG, Hara Y (1998) Antioxidant properties of flavonoids: reduction potentials and electron transfer reactions of flavonoid radicals. In: Rice-Evans C, Packer JE (eds) Flavonoids in health and disease. Marcel Dekker, Inc, New York, pp 137–161Google Scholar
  38. 38.
    Kandaswami C, Perkins E, Soloniuk DS, Drzewiecki G, Middleton EJ (1993) Ascorbic acid-enhanced antiproliferative effect of flavonoids on squamous cell carcinoma in vitro. Anticancer Drugs 4:91–96PubMedGoogle Scholar
  39. 39.
    Jovanovic SV, Steenken S, Tosic M, Marjanovic B, Simic MG (1994) Flavonoids as antioxidants. J Am Chemical Society 116:4846–4851CrossRefGoogle Scholar
  40. 40.
    Steenken S, Neta P (1982) One-electron redox potentials of phenols. Hydroxyand aminophenols and related compounds of biological interest. J Physical Chemistry 86:3661–3667Google Scholar
  41. 41.
    Buettner GR, Jurkiewicz BA (1993) Ascorbate free radical as a marker of oxidative stress: an EPR study. Free Radic Biol Med 14:49–55PubMedGoogle Scholar
  42. 42.
    Frei B, Keaney JF, Retsky KL, Chen K (1996) Vitamins C and E and LDL oxidation. Vitam Horm 52:1–34PubMedGoogle Scholar
  43. 43.
    Esterbauer H, Dieber-Rotheneder M, Striegl G, Waeg G (1991) Role of vitamin E in preventing the oxidation of lowdensity lipoprotein. Am J Clin Nutr 53:314S–321SPubMedGoogle Scholar
  44. 44.
    Terao J, Piskula M (1997) Flavonoids as inhibitors of lipid peroxidation in membranes. In: Rice-Evans C, Packer JE (eds) Flavonoids in health and disease. Marcel Dekker Inc, New York, pp 277–294Google Scholar

Copyright information

© Steinkopff-Verlag 2005

Authors and Affiliations

  1. 1.Dept. of Food and Nutrition TechnicalUniversity of MunichFreising-WeihenstephanGermany
  2. 2.MunichGermany
  3. 3.Unit of Human Nutrition and Cancer PreventionTechnical University of MunichMunichGermany

Personalised recommendations