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Evaluation of the Genoprotective Effect of Polyphenolic, Hydroalcoholic and Aqueous Extracts of Red Apple
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Apple has a tremendous supply of minerals, vitamins C, B Group and phenolic compounds. It has antioxidant properties and maintains genetic content against oxidative stress. In animal experiments, methyl methanesulfonate (MMS) has demonstrated genotoxic effects. In this study, the genoprotective effects versus the MMS genotoxicity of red apple extracts were investigated. Red apples (Red Delicious cultivar) were chopped in thin slices and three types of extracts consisting polyphenolic, hydroalcoholic and aqueous were taken. Lymphocytes were pre-incubated with various concentrations of extracts and then were exposed to 10μM of MMS. This was made possible by using alkaline comet assay and analysis of comet images. The total phenolic content (TPC) for polyphenolic, hydroalcoholic and aqueous extracts were obtained 127.74 ± 4.26, 118.12 ± 4.47 and 84.79 ± 4.26 mg/g, respectively. The extracts consisted of 48.92 ± 1.02, 42.09 ± 3.36, 29.08 ± 3.23 mg/g of total flavonoid (TF). The results illustrated that in genoprotective concentrations of 1, 10 and 100μg/ml for all these extracts, the tail length and tail moment variables were significantly lower than the positive control (10μM of MMS) (p< 0.05). In this study, results revealed that these extracts were able to protect lymphocytes against MMS. By considering the TPC and TF values, the polyphenolic and hydroalcoholic extracts had more genoprotective activity compared to the aqueous extract. The polyphenolic extract was proved to be the most protective extract of all.
Keywords
Comet Assay, DNA Damage, Methyl Methanesulfonate, Malus domestica.
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- De Bont R, van Larebeke N. Endogenous DNA damage in humans: a review of quantitative data. Mutagenesis. 2004; 19:169–85.
- Cadet J, Delatour T, Douki T, Gasparutto D, Pouget JP, Ravanat JL, et al. Hydroxyl radicals and DNA base damage. Mutat Res. 1999; 424:9–21.
- Valerie K, Povirk LF. Regulation and mechanisms of mammalian double-strand break repair. Oncogene. 2003; 22:5792–812.
- Ferguson LR, Denny WA. The genetic toxicology of acridines. Mutat Res. 1991; 258:123–60.
- D’Argenio G, Mazzone G, Tuccillo C, Ribecco MT, Graziani G, Gravina AG, et al. Apple polyphenols extract (APE) improves colon damage in a rat model of colitis. Dig Liver Dis. 2012; 44:555–62.
- Chen C-S, Zhang D, Wang Y-Q, Li P-M, Ma F-W. Effects of fruit bagging on the contents of phenolic compounds in the peel and flesh of ‘Golden Delicious’, ‘Red Delicious’, and ‘Royal Gala’ apples. Scientia Horticulturae. 2012; 142:68– 73.
- Fu L, Xu B-T, Xu X-R, Gan R-Y, Zhang Y, Xia E-Q, et al. Antioxidant capacities and total phenolic contents of 62 fruits. Int J Mol Sci. 2011; 129:345–50.
- Lam CK, Zhang Z, Yu H, Tsang S-Y, Huang Y, Chen ZY. Apple polyphenols inhibit plasma CETP activity and reduce the ratio of non-HDL to HDL cholesterol. Mol Nutr Food Res. 2008; 52:950–8.
- Miene C, Klenow S, Veeriah S, Richling E, Glei M. Impact of apple polyphenols on GSTT2 gene expression, subsequent protection of DNA and modulation of proliferation using LT97 human colon adenoma cells. Molecular Nutrition and Food Research. 2009; 53:1254–62.
- Veeriah S, Miene C, Habermann N, Hofmann T, Klenow S, Sauer J, et al. Apple polyphenols modulate expression of selected genes related to toxicological defence and stress response in human colon adenoma cells. International Journal of Cancer. 2008; 122:2647–55.
- McCann MJ, Gill CIR, O’ Brien G, Rao JR, McRoberts WC, Hughes P, et al. Anti-cancer properties of phenolics from apple waste on colon carcinogenesis in vitro. Food Chem Toxicol. 2007; 45:1224–30.
- Cheynier V. Polyphenols in foods are more complex than often thought. Am J Clin Nutr. 2005; 81:223S–9S.
- ICH working group. Guidelines on genotoxicity testing and data interpretation for pharmaceutical intended for human use, S2R1. 2011 Nov 9. Available from: http://www.fda.gov/ downloads/Drugs/Guidances/ucm074931.pdf
- Tice RR, Agurell E, Anderson D, Burlinson B, Hartmann A, Kobayashi H, et al. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environ Mol Mutagen. 2000; 35:206–21.
- Collins AR. The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol. 2004; 26:249–61.
- Hajhashemi V, Ghannadi A, Pezeshkian SK. Antinociceptive and anti-inflammatory effects of Satureja hortensis L extracts and essential oil. J Ethnopharmacol. 2002; 82:83–7.
- Sahu R, Saxena J. Screening of Total Phenolic and Flavonoid Content in Conventional and Non-Conventional Species of Curcuma. Journal of Pharmacognosy and Phytochemistry. 2013; 2:176–9.
- English D, Andersen BR. Single-step separation of red blood cells, granulocytes and mononuclear leukocytes on discontinuous density gradients of Ficoll-Hypaque. J Immunol Methods. 1974; 5:249–52.
- Kalmar JR, Arnold RR, Warbington ML, Gardner MK. Superior leukocyte separation with a discontinuous one-step Ficoll-Hypaque gradient for the isolation of human neutrophils. J Immunol Methods. 1988; 110:275–81.
- Collins AR, Oscoz AA, Brunborg G, Gaivao I, Giovannelli L, Kruszewski M et al. The comet assay: topical issues. Mutagenesis. 2008; 23:143–51.
- Brendler-Schwaab S, Hartmann A, Pfuhler S, Speit G. The in vivo comet assay: use and status in genotoxicity testing. Mutagenesis. 2005; 20:245–54.
- Vieira FGK, Borges GDSC, Copetti C, Amboni RDDMC, Denardi F, Fett R. Physico-chemical and antioxidant properties of six apple cultivars (Malus domestica Borkh) grown in southern Brazil. Scientia Horticulturae. 2009; 122:421–5.
- Lodish H, Berk A, Zipursky SL et al. Molecular Cell Biology. 4th edition. New York: W. H. Freeman; 2000. Section 12.4, DNA Damage and Repair and Their Role in Carcinogenesis. Available from: http://www.ncbi.nlm.nih.gov/books/NBK21554/
- Carrasco-Pozo C, Gotteland M, Speisky H. Protection by apple peel polyphenols against indometacin-induced oxidative stress, mitochondrial damage and cytotoxicity in Caco-2 cells. J Pharm Pharmacol. 2010; 62:943–50.
- Veeriah S et al. 2006. Apple flavonoids inhibit growth of HT29 human colon cancer cells and modulate expression of genes involved in the biotransformation of xenobiotics. Molecular Carcinogenesis. 45: 164–74.
- Barth SW, Fahndrich C, Bub A, Dietrich H, Watzl B, Will F et al. Cloudy apple juice decreases DNA damage, hyperproliferation and aberrant crypt foci development in the distal colon of DMH-initiated rats. Carcinogenesis. 2005; 26(8):1414–21.
- Boyer J, Liu RH. Apple phytochemicals and their health benefits. Nutrition Journal. 2004; 3(5):111–5.
- Collins AR. The comet assay for DNA damage and repair: principles, applications, and limitations. Mol Biotechnol. 2004; 26:249–61.
- Schaefer S, Baum M, Eisenbrand G, Dietrich H, Will F, Janzowski C. Polyphenolic apple juice extracts and their major constituents reduce oxidative damage in human colon cell lines. Mol Nutr Food Res. 2006; 50:24–33.
- Bausinger J, Speit G. DNA repair capacity of cultured human lymphocytes exposed to mutagens measured by the comet assay and array expression analysis. Mutagenesis. 2015.
- Faust F, Kassie F, Knasmuller S, Boedecker RH, Mann M, Mersch-Sundermann V. The use of the alkaline comet assay with lymphocytes in human biomonitoring studies. Mutat Res. 2004; 566(3):209–29. Epub 2004/04/15.
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