Antioxidant Properties of Grape Seed Extract on Human Lymphocyte Oxidative Defence

Abstract

The distribution of polyphenolic compounds in a grape (Vitis vinifera) seed extract (GSE) was determined using LC/ESI-TOF MS, HPLC/DAD, and ¹³C-NMR. The 17 identified compounds comprised gallic and protocatechuic acid, catetchin and epicatechin monomers, procyanidin oligomers, and procyanidin gallates. This study addresses the in vitro effects of grape seed extract (GSE) on the frequency of micronuclei with reference to the antioxidant status in human lymphocytes. To establish the most effective protective support, we used four different concentrations of GSE, in the range 1 - 6 μg/mL. Treatment of lymphocytes with GSE at a concentration of 2.5 μg/mL induced a significant decrease in the frequency of micronuclei by 40 %, reduction of malonyldialdehyde production by 30 %, while a concentration of 5 μg/mL increased catalase and glutathione S-transferase activity by 10 % and 15 %, respectively. These results demonstrate that GSE may be effective in the prevention of oxidative lymphocyte damage by ROS.

Key words

Vitis vinifera - Vitaceae - micronucleus test - antioxidant enzymes - human lymphocytes - LC/MS - NMR

References

• 1 Tominaga H, Kodama S, Matsuda N, Suzuki K, Watanabe M. Involvement of reactive oxygen species (ROS) in the induction of genetic instability by radiation.  J Radiat Res. 2004;  45 181-8
• 2 Prasad N, Srinivasan M, Pugalendi V K, Menon P V. Protective effect of ferulic acid on δ-radiation-induced micronuclei, dicentric aberration and lipid peroxidation in human lymphocytes.  Mutat Res. 2006;  603 129-34
• 3 Ishige K, Schubert D, Sagara Y. Flavonoids protect neuronal cells from oxidative stress by three distinct mechanisms.  Free Radic Biol Med. 2001;  30 433-46
• 4 Ma W, Nunes I, Young H SC, Spector A. Catalase enrichment using recombinant adenovirus protects αTN4 - 1 cells from H₂O₂.  Free Radic Biol Med. 2006;  40 335-40
• 5 Hunt C, Sim J E, Sullivan S J, Featherstone T, Golden W, Kapp-Herr C V. et al . Genomic instability and catalase gene amplification induced by chronic exposure to oxidative stress.  Cancer Res. 1998;  58 3986-92
• 6 Fenech M, Morley A A. Measurement of micronuclei in lymphocytes.  Mutat Res. 1985;  147 29-36
• 7 Pekić B, Kovač V, Alonso E, Revilla E. Study of the extraction of proanthocyanidins from grape seeds.  Food Chem. 1998;  61 201-6
• 8 Fenech M, Morley A A. The cytokinesis-block micronucleus technique: A detailed description of the method and its application to genotoxicity studies in human population.  Mutat Res. 1993;  285 35-44
• 9 Countryman P I, Heddle J A. The production of micronuclei from chromosome aberration in irradiated cultures of human lymphocytes.  Mutat Res. 1976;  41 321-32
• 10 Arouma O I, Halliwell B, Laughton M J, Quinlan G J, Gutteridge J M. The mechanism of initiation of lipid peroxidation. Evidence against a requirement for an iron (II)-iron(III) complex.  Biochem J. 1989;  258 617-20
• 11 Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.  Anal Biochem. 1976;  72 248-54
• 12 Habig H W, Michael J P, Jakoby B W. Glutathione S-transferases, the first enzymatic step in mercapturic acid formation.  J Biol Chem. 1974;  249 7130-9
• 13 Aebi H. Catalase in vitro. .  Methods Enzymol. 1984;  105 121-6
• 14 Newman R H, Porter L J, Foo L Y. High-resolution ¹³C NMR studies of proanthocyanidin polymers (condensed tannins).  Magn Reson Chem. 1987;  25 118-24
• 15 Behrens A, Maie N, Knicker H, Kogel-Knabner I. MALDI TOF mass spectrometry and PSD fragmentation as means for the analysis of condensed tannins in plant leaves and needles.  Phytochemistry. 2003;  62 1159-70
• 16 Ohshima H, Yoshie Y, Auriol S, Gilibert I. Antioxidant and pro-oxidant actions of flavonoids: effects on DNA damage induced by nitric oxide, peroxynitrite and nitroxyl anion.  Free Radic Biol Med. 1998;  25 1057-65
• 17 Sirinivasan P, Vadhanam V M, Arif M J, Gupta C R. A rapid screening assay for antioxidant potential of natural and synthetic agents in vitro. .  Int J Oncol. 2002;  20 983-6
• 18 Teissedre P L, Waterhouse A L, Frankel E N. Principal phenolic phytochemicals in French Syral and Grenache Rhone wines and their antioxidant activity in inhibiting oxidation of human low density lipoproteins.  J Int Sci Vigne Vin. 1995;  29 205-12
• 19 Fenech M, Stockley C, Aitken C. Moderate wine consumption protects against hydrogen peroxide-induced DNA damage.  Mutagenesis. 1997;  12 289-96
• 20 Shao Z H, Qin Y M, Becker B L, Vanden Hoek T L, Li C Q, Schumacker T P. et al . Grape proanthocyanidins reduce oxidant stress in cardiomyocytes.  Acad Emerg Med. 2001;  8 562-8
• 21 Müller A C, Pigorsch S, Beyer C, Lautenschläger C, Dunst J. Radioprotective effects of amifostine in vitro and in vivo measured with the comet assay.  Strahlenther Oncol. 2004;  180 517-25