Comparative effects of dietary flavanols on antioxidant defences and their response to oxidant-induced stress on Caco2 cells

  1. Ildefonso Rodríguez-Ramiro 1
  2. María Ángeles Martín 1
  3. Sonia Ramos 1
  4. Laura Bravo 1
  5. Luis Goya 1
  1. 1 Department of Metabolism and Nutrition, Instituto del Frío-ICTAN, CSIC, José Antonio Novais 10, Ciudad Universitaria, 28040, Madrid, Spain
Revista:
European Journal of Nutrition

ISSN: 1436-6207,  1436-6215

Año de publicación: 2010

Volumen: 50

Número: 5

Páginas: 313-322

Tipo: Artículo

DOI: 10.1007/S00394-010-0139-2 GOOGLE SCHOLAR lock_openAcceso abierto editor

Otras publicaciones en: European Journal of Nutrition

Resumen

Purpose Flavanols are an important fraction of our dietboth for their antioxidant capacity and because they areconstituents of greatly accepted foodstuffs such as tea,wine and cocoa. In addition to their antioxidant activity bydirectly scavenging intracellular reactive oxygen species(ROS), flavanols have been recently shown to enhanceprotective enzymes. The objective was to evaluate theantioxidant response of colon-derived Caco2 cells to dietary flavanols.Methods Four representative flavanols were selected:epicatechin (EC), epicatechin-3-gallate (ECG), epigallocatechin-3-gallate (EGCG) and procyanidin B2 (PB2). Cellviability, concentration of ROS and reduced glutathione(GSH), and activity of antioxidant/detoxification enzymesand caspase 3 were determined.Results Treatment of Caco2 cells with flavanolsdecreased ROS production but did not affect GSH content.ECG induced glutathione peroxidase (GPx), whereas PB2evoked a dose-dependent increase in GPx, glutathionereductase and glutathione-S-transferase. Enhancement ofthe antioxidant defences implies an improved cell responseto an oxidative challenge. Hence, Caco2 cells treated 20 hwith the flavanols, especially PB2, and then submittedto an oxidative stress induced by a pro-oxidant, tert-butylhydroperoxide, showed a reduced ROS production, restricted activation of caspase 3 and higher viability thancells plainly submitted to the stressor.Conclusions Flavanols protect Caco2 cells against aninduced oxidative stress and subsequent cellular death byreducing ROS production and preventing caspase-3 activation. In particular, PB2 increases the activity of antioxidant/detoxification enzymes and thus protects Caco2cells by directly counteracting free radicals and also byactivating the antioxidant defence system.

Referencias bibliográficas

  • Sohal RS, Mockett RJ, Orr WC (2002) Mechanisms of aging. An appraisal of the oxidative stress hypothesis. Free Radic Biol Med 33:575–586
  • Ramos S (2008) Cancer chemoprevention and chemotherapy: dietary polyphenols and signalling pathways. Mol Nutr Food Res 52:507–526
  • Mukhatar H, Ahmad N (2000) Tea polyphenols: prevention of cancer and optimizing health. Am J Clin Nutr 71:1698S–1702S
  • Ahn WS, Yoo J, Huh SW, Kim CK, Lee JM, Namkoong SE, Bae SM, Lee IP (2003) Protective effects of green tea extracts (polyphenon E and EGCG) on human cervical lesions. Eur J Cancer Prev 12:383–390
  • Bettuzzi S, Rizzi F, Belloni L (2007) Clinical relevance of the inhibitory effect of green tea catechins (GtCs) on prostate cancer progression in combination with molecular profiling of catechin-resistant tumors: an integrated view. Pol J Vet Sci 10:57–60
  • Martín MA, Ramos S, Mateos R, Granado-Serrano AB, Izquierdo-Pulido M, Bravo L, Goya L (2008) Protection of human HepG2 cells against oxidative stress by cocoa phenolic extract. J Agric Food Chem 56:7765–7772
  • Martín MA, Ramos S, Mateos R, Izquierdo-Pulido M, Bravo L, Goya L (2010) Protection of human HepG2 cells against oxidative stress induced by the flavonoid epicatechin. Phytother Res 24:503–509
  • Masella R, Di Benedetto R, Varı R, Filesi C, Giovannini C (2005) Novel mechanisms of natural antioxidant compounds in biological systems: involvement of glutathione and glutathione-related enzymes. J Nutr Biochem 16:577–586
  • Na HK, Surh YJ (2008) Modulation of Nrf2-mediated antioxidant and detoxifying enzyme induction by the green tea polyphenol EGCG. Food Chem Toxicol 46:1271–1278
  • Alía M, Mateos R, Ramos S, Lecumberri E, Bravo L, Goya L (2006) Influence of quercetin and rutin on growth and the antioxidant defense system in a human hepatoma cell line (HepG2). Eur J Nutr 45:19–28
  • Molina MF, Sanchez-Reus I, Iglesias I, Benedi J (2003) Quercetin, a flavonoid antioxidant, prevents and protects against ethanol induced oxidative stress in mouse liver. Biol Pharm Bull 26:1398–1402
  • Martín MA, Granado-Serrano AB, Ramos S, Izquierdo-Pulido M, Bravo L, Goya L (2010) Cocoa flavonoids up-regulate antioxidant enzymes activity via ERK1/2 pathway to protect against oxidative stress-induced apoptosis in HepG2 cells. J Nutr Biochem 21:196–205
  • Baba S, Osakabe N, Natsume N, Muto Y, Takizawa T, Terao J (2001) In vivo comparison of the bioavailability of catechin, epicatechin and their mixture in orally administered rats. J Nutr 131:2885–2891
  • Sambuy Y, De Angelis I, Ranaldi G, Scarino ML, Stammati A, Zucco F (2005) The Caco2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco2 cell functional characteristics. Cell Biol Toxicol 21:1–26
  • Rufián-Henares JA, Morales FJ (2007) Effect of in vitro enzymatic digestion on antioxidant activity of coffee melanoidins and fractions. J Agric Food Chem 55:10016–10021
  • Martín MA, Ramos S, Mateos R, Rufián-Henares JA, Morales FJ, Bravo L, Goya L (2009) Biscuit melanoidins of different molecular masses protect human HepG2 cells against oxidative stress. J Agric Food Chem 57:7250–7258
  • Welder AA, Acosta D (1994) Enzyme leakage as an indicator of cytotoxicity in culture cells. In: Tyson CA, Franzier JM (eds) In vitro toxicity indicators: methods in toxicology. Academic Press, New York, pp 46–49
  • Wang H, Joseph JA (1999) Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radic Biol Med 27:612–616
  • Hissin PJ, Hilf R (1976) A fluorometric method for determination of oxidised and reduced glutathione in tissues. Anal Biochem 74:214–226
  • Gunzler WA, Kramers H, Flohe L (1974) An improved coupled test procedure for glutathione peroxidase. Klin Chem Klin Biochem 12:444
  • Goldberg DM, Spooner RJ (1987) Glutathione reductase. In: Bergmeyer HV (ed) Methods of enzymatic analysis. Verlag-Chemie, Weinheim, pp 258–265
  • Herrera B, Fernández M, Alvarez A, Roncero C, Benito M, Gil J, Fabregat I (2001) Activation of caspases occurs downstream from radical oxygen species production, Bcl-xL down-regulation, and early cytochrome C release in apoptosis induced by transforming growth factor beta in rat fetal hepatocytes. Hepatology 34:548–556
  • Steffen Y, Jung T, Klotz L-O, Schewe T, Grune T, Sies H (2007) Protein modification elicited by oxidized LDL in endothelial cells: protection by epicatechin. Free Radic Biol Med 42:955–970
  • Natsume M, Osakabe N, Yasuda A, Baba S, Tokunaga T, Kondo K, Osawa T, Terao J (2004) In vitro antioxidative activity of epicatechin glucuronide metabolites present in human and rat plasma. Free Radic Res 38:1341–1348
  • Jourdain C, Tenca G, Deguercy A, Troplin P, Poelman D (2006) In vitro effects of polyphenols from cocoa and beta sitosterol on the growth of human prostate cancer and normal cells. Eur J Cancer Prev 4:353–361
  • Ramiro E, Franch A, Castellote C, Pérez-Cano F, Permanyer J, Izquierdo-Pulido M, Castell M (2005) Flavonoids from Theobroma cacao down-regulate inflammatory mediators. J Agric Food Chem 53:8506–8511
  • Rice-Evans C (2004) Flavonoids and isoflavones: absorption, metabolism and bioactivity. Free Radic Biol Med 36:827–828
  • Chang W-T, Shao Z-H, Yin J-J, Mehendale S, Wang CZ, Qin Y, Li J, Chen WJ, Chien CT, Becker LB, Van den Hoek TL, Yuan CS (2007) Comparative effects of flavonoids on oxidant scavenging and ischemia-reperfusion injury cardiomyocytes. Eur J Pharmacol 566:58–66
  • Granado-Serrano AB, Martín MA, Izquierdo-Pulido M, Goya L, Bravo L, Ramos S (2007) Molecular mechanisms of (−)-epicatechin and chlorogenic acid on the regulation of the apoptotic and survival/proliferation pathways in a human hepatoma cell line (HepG2). J Agric Food Chem 55:2020–2027
  • Arts IC, van de Putte B, Hollman PC (2000) Catechin contents of foods commonly consumed in The Netherlands. 1. Fruits, vegetables, staple foods and processed foods. J Agric Food Chem 48:1746–1751
  • De Pascual-Teresa S, Santos-Buelga C, Rivas-Gozalo JC (2000) Quantitative analysis of flavan-3-ols in Spanish foodstuffs and beverages. J Agric Food Chem 48:5331–5337
  • Appeldoorn MM, Vincken JP, Gruppen H, Hollman PC (2009) Procyanidin dimers A1, A2 and B2 are absorbed without conjugation or methylation from the small intestine of rats. J Nutr 139:1469–1473
  • Luximon-Ramma A, Bahorun T, Crozier A, Zbarsky V, Datla KP, Dexter DT, Aruoma OI (2005) Characterization of the antioxidant functions of flavonoids and proanthocyanidins in Mauritian black teas. Food Res Int 38:357–367
  • Soobrattee MA, Neergheen VS, Luximon-Ramma A, Aruoma OI, Bahorun T (2005) Phenolics as potential antioxidant therapeutic agents. Mutat Res 579:200–213
  • Scharf G, Prustomersky S, Knasmuller S, Schulte-Hermann R, Huber WW (2003) Enhancement of glutathione and g-glutamylcysteine synthetase, the rate limiting enzyme of glutathione synthesis, by chemoprotective plant-derived food and beverage components in the human hepatoma cell line HepG2. Nutr Cancer 45:74–83
  • Lei XG, Cheng W-H, McClung JP (2007) Metabolic regulation and function of glutathione peroxidase-1. Annu Rev Nutr 27:41–61
  • Argyrou A, Blanchard JS (2004) Flavoprotein disulfide reductases: advances in chemistry and function. Prog Nucleic Acid Res Mol Biol 78:89–142
  • Masella R, Varı R, D’Archivio M, Di Benedetto R, Mataresse P, Malorni W, Scazzocchio B, Giovannini C (2004) Extra virgin olive oil biophenols inhibit cell-mediated oxidation of LDL by increasing the mRNA transcription of glutathione-related enzymes. J Nutr 134:785–791
  • Ramiro Puig E, Urpí-Sardá M, Pérez-Cano FJ, Franch A, Castellote C, Andrés-Lacueva C, Izquierdo-Pulido M, Castell M (2008) Cocoa-enriched diet enhances antioxidant enzyme activity and modulates lymphocyte composition in thymus from young rats. J Agric Food Chem 55:6431–6438
  • Chen C, Kong AN (2004) Dietary cancer-chemopreventive compounds: from signaling and gene expression to pharmacological effects. Free Radic Biol Med 36:1505–1516
  • Singh R, Czaja MJ (2007) Regulation of hepatocyte apoptosis by oxidative stress. Gastroenterol Hepatol 1:S45–S48
  • De Ruvo C, Amodio R, Algeri S, Martelli N, Intilangelo A, D′Ancona GM, Esposito E (2000) Nutritional antioxidants as antidegenerative agents. Int J Dev Neurosci 18:359–366
  • Granado-Serrano AB, Martín MA, Goya L, Bravo L, Ramos S (2009) Time course regulation of survival pathways by epicathechin on HepG2. J Nutr Biochem 20:115–124