Whey protein, as exclusively nitrogen source, controls food intake and promotes glutathione antioxidant protection in Sprague-Dawley rats

  • Samir G. Sukkar
  • Franca Cella
  • Stefania Patriarca
  • Anna L. Furfaro
  • Francesca Abate
  • Claudia Ferrari
  • Emanuela Balbis
  • Nicola Traverso
  • Damiano Cottalasso
Original Article


The inclusion of whey protein concentrates (WPC) in the diet can lead to a decrease in food intake. Considering that excessive food intake and weight gain are correlated with increased oxidative stress and other risk factors, the anorectic action of WPC may have important clinical implications. The aims of the current study were to verify the effects of WPC in comparison with those of casein on food intake, weight, and oxidized glutathione (GSSG) and total glutathione (GSH) concentrations in the blood and liver with or without oxidative stress induced by oral carbon tetrachloride intoxication. Male Sprague-Dawley rats were fed a balanced liquid diet for 3 weeks. Half of the rats received WPC (group P), while the control group received casein (group C). Group P rats ate significantly less than group C rats (p < 0.0001), and their weights decreased significantly. After carbon tetrachloride intoxication, there was a significant increase in GSH in rats of group P compared with the levels in rats of group C both in the liver (GSH group P 4,994 ± 652.6, group C 2,196 ± 323.2 nmol/mg, p < 0.01) and in the blood (GSH group P 1,368 ± 69.56, group C 1,088 ± 48.35 nmol/ml, p < 0.05). These findings indicate that WPC is effective in reducing food intake and preventing weight gain, and it may also play a protective role against oxidative stress by increasing glutathione synthesis in the liver.


Whey protein Glutathione Appetite inhibition Energy intake Antioxidant defences Oxidative stress Acute carbon tetrachloride intoxication 


  1. 1.
    Jean C, Rome S, Mathe V et al (2001) Metabolic evidence for adaptation to a high protein diet in rats. J Nutr 131:91–98Google Scholar
  2. 2.
    Bensaid A, Tome D, L’Heureux-Bourdon D et al (2003) A high-protein diet enhances satiety without conditioned taste aversion in the rat. Physiol Behav 78:311–320CrossRefGoogle Scholar
  3. 3.
    Hall WL, Millward DJ, Long SJ, Morgan LM (2003) Casein and whey exert different effects on plasma amino acid profiles, gastrointestinal hormone secretion and appetite. Br J Nutr 89:239–248CrossRefGoogle Scholar
  4. 4.
    Boirie Y, Dangin M, Gachon P et al (1997) Slow and fast dietary proteins differently modulate postprandial protein accretion. Proc Natl Acad Sci U S A 94:14930–14935CrossRefGoogle Scholar
  5. 5.
    Fruhneck G (1998) Slow and fast dietary proteins. Nature 391:843–844CrossRefGoogle Scholar
  6. 6.
    Asensi M, Sastre J, Pallardo FV et al (1999) Ratio of reduced to oxidized glutathione as indicator of oxidative stress status and DNA damage. Methods Enzymol 299:267–276CrossRefGoogle Scholar
  7. 7.
    McIntosh GH, Regester GQ, Le Leu RK, Royle PJ (1995) Dairy proteins protect against dimethylhydrazine-induced intestinal cancers in rats. J Nutr 125:809–816Google Scholar
  8. 8.
    Dröge W (2002) Free radicals in the physiological control of cell function. Physiol Rev 82:47–95Google Scholar
  9. 9.
    McIntosh GH, Jorgensen L, Royle P (1993) The potential of an insoluble dietary fiber-rich source from barley to protect from DMH-induced intestinal tumours in rats. Nutr Cancer 19:213–221Google Scholar
  10. 10.
    Pacheco MT, Sgarbieri VC (2005) Effect of different hydrolysates of whey protein on hepatic glutathione content in mice. J Med Food 8:337–342CrossRefGoogle Scholar
  11. 11.
    Lands LC, Grey VL, Smountas AA (1999) Effects of supplementation with a cysteine donor on muscular performance. J Appl Physiol 87:1381–1385Google Scholar
  12. 12.
    Belobrajdic DP, McIntosh GH, Owens JA (2004) A high-wheyprotein diet reduces body weight gain and alters insulin sensitivity relative to red meat in Wistar rats. J Nutr 134:1454–1458Google Scholar
  13. 13.
    Keaney JF Jr, Larson MG, Vasan RS et al (2003) Framingham Study. Obesity and systemic oxidative stress: clinical correlates of oxidative stress in the Framingham Study. Arterioscler Thromb Vasc Biol 23:434–439CrossRefGoogle Scholar
  14. 14.
    Roskams T, Yang SQ, Koteish A et al (2003) Oxidative stress and oval cell accumulation in mice and humans with alcoholic and non-alcoholic fatty liver disease. Am J Pathol 163:1301–1311Google Scholar
  15. 15.
    Olusi SO (2002) Obesity is an independent risk factor for plasma lipid peroxidation and depletion of erythrocyte cytoprotective enzymes in humans. Int J Obes Relat Metab Disord 26:1159–1164CrossRefGoogle Scholar
  16. 16.
    Hensley K, Floyd RA (2002) Reactive oxygen species and protein oxidation in aging: a look back, a look ahead. Arch Biochem Biophys 397:377–383CrossRefGoogle Scholar
  17. 17.
    Mariotti F, Simbelie KL, Makarios-Lahham L et al (2004) Acute ingestion of dietary proteins improves post-exercise liver glutathione in rats in a dose-dependent relationship with their cysteine content. J Nutr 134:128–131Google Scholar
  18. 18.
    Ritter C, Reinke A, Andrades M et al (2004) Protective effect of N-acetylcysteine and deferoxamine on carbon tetrachloride-induced acute hepatic failure in rats. Crit Care Med 32:2079–2083CrossRefGoogle Scholar
  19. 19.
    Dwivedi S, Sharma R, Sharma A et al (2006) The course of CCl4 induced hepatotoxicity is altered in mGSTA4-4 null (-/-) mice. Toxicology 218:58–66CrossRefGoogle Scholar
  20. 20.
    Aneja R, Upadhyaya G, Prakash S et al (2005) Ameliorating effect of phytoestrogens on CCL4-induced oxidative stress in the livers of male Wistar rats. Artif Cells Blood Substit Immobil Biotechnol 33:201–213CrossRefGoogle Scholar
  21. 21.
    Basu S (2003) Carbon tetrachloride-induced lipid peroxidation: eicosanoid formation and their regulation by antioxidant nutrients. Toxicology 189:113–127CrossRefGoogle Scholar
  22. 22.
    Lieber CS, Decarli LM (1989) Liquid diet technique of ethanol administration: 1989 update. Alcohol Alcohol 24:197–211Google Scholar
  23. 23.
    Reeves PG, Nielsen FH. Fahey GC (1993) AIN-93 purified diets for laboratory rodents: final report al the American Institute of Nutrition ad hoc writing committee on the reformulation of the AIN-76A rodent diet. J Nutr 123:1939–1951Google Scholar
  24. 24.
    Fariss MW, Reed DJ (1987) High-performance liquid chromatography of thiols and disulfides: dinitrophenol derivatives. Methods Enzymol 143:101–109CrossRefGoogle Scholar
  25. 25.
    Furukawa S, Fujita T, Shimabukuro M et al (2004) Increased oxidative stress in obesity and its impact on metabolic syndrome. J Clin Invest 114:1752–1761Google Scholar
  26. 26.
    Rossi R, Milzani A, Dalle-Donne I et al (2002) Blood glutathione disulfide: in vivo factor or in vitro artifact? Clin Chem 48:742–753Google Scholar
  27. 27.
    Noelle RJ, Lawrence DA (1981) Determination of glutathione in lymphocytes and possible association of redox state and proliferative capacity of lymphocytes. Biochem J 198:571–579Google Scholar
  28. 28.
    Feinle C, O’Donovan D, Doran S et al (2003) Effects of fat digestion on appetite, APD motility, and gut hormones in response to duodenal fat infusion in humans. Am J Physiol Gastrointest Liver Physiol 284:G798–G807Google Scholar
  29. 29.
    Dangin M, Boirie Y, Guillet C, Beaufrere B (2002) Influence of the protein digestion rate on protein turnover in young and elderly subjects. J Nutr 132:3228S–3233SGoogle Scholar
  30. 30.
    Bowen J, Noakes M, Clifton P et al (2004) Effect of dietary proteins on appetite, energy intake and glycemic response in overweight men. Asia Pac J Clin Nutr 13[Suppl]:S64Google Scholar
  31. 31.
    Hum S, Koski KG, Hoffer LJ (1992) Varied protein intake alters glutathione metabolism in rats. J Nutr 122:2010–2018Google Scholar
  32. 32.
    Dröge W (1999) Cysteine and glutathione in catabolic conditions and immunological dysfunction. Curr Opin Clin Nutr Metab Care 2:227–233CrossRefGoogle Scholar
  33. 33.
    Parker NT, Goodrum KJ (1990) A comparison of casein, lactalbumin, and soy protein effect on the immune response to a Tdependent antigen. Nutr Res 10:781–792CrossRefGoogle Scholar
  34. 34.
    Wong CW, Watson DL (1995) Immunomodulatory effects of dietary whey proteins in mice. J Dairy Res 62:350–368CrossRefGoogle Scholar
  35. 35.
    Bounous G, Létourneau L, Kongshavn PA (1983) Influence of dietary protein type on the immune system of mice. J Nutr 113:1415–1421Google Scholar
  36. 36.
    Bounous G, Batist G, Gold P (1989) Immunoenhancing property of dietary whey protein in mice: role of glutathione. Clin Invest Med 12:154–161Google Scholar
  37. 37.
    Ames BN, Shigenaga MK, Hagen TM (1993) Oxidants, antioxidants, and the degenerative diseases of aging. Proc Natl Acad Sci U S A 90:7915–7922CrossRefGoogle Scholar
  38. 38.
    Beckman KB, Ames BN (1998) The free radical theory of aging matures. Physiol Rev 78:547–581Google Scholar
  39. 39.
    Lee CK, Klopp RG, Weindruch R, Prolla TA (1999) Gene expression profile of aging and its retardation by caloric restriction. Science 285:1390–1393CrossRefGoogle Scholar
  40. 40.
    Lee CK, Weindruch R, Prolla TA (2000) Gene expression profile of the aging brain in mice. Nat Genet 25:294–297CrossRefGoogle Scholar
  41. 41.
    Sohal RS, Weindruch R (1996) Oxidative stress, caloric restriction, and ageing. Science 273:59–63CrossRefGoogle Scholar
  42. 42.
    Zainal TA, Oberley TD, Allison DB et al (2000) Caloric restriction of rhesus monkeys lowers oxidative damage in skeletal muscle. FASEB J 14:1825–1836CrossRefGoogle Scholar
  43. 43.
    Hack V, Breitkreutz R, Kinscherf R et al (1998) The redox state as a correlate of senescence and wasting and as a target for therapeutic intervention. Blood 92:59–67Google Scholar
  44. 44.
    Ji LL, Dillon D, Wu E (1990) Alteration of antioxidant enzymes with aging in rat skeletal muscle and liver. Am J Physiol. 258:R918–R923Google Scholar
  45. 45.
    Dangin M, Guillet C, Garcia-Rodenas C et al (2003) The rate of protein digestion affects protein gain differently during aging in humans. J Physiol 549:635–644CrossRefGoogle Scholar
  46. 46.
    Miquel J, Ferrandiz ML, de Juan E et al (1995) N-acetylcysteine protects against age-related decline of oxidative phosphorylation in liver mitochondria. Eur J Pharmacol 292:333–335Google Scholar

Copyright information

© Springer-Verlag Italia 2008

Authors and Affiliations

  • Samir G. Sukkar
    • 1
  • Franca Cella
    • 1
  • Stefania Patriarca
    • 2
  • Anna L. Furfaro
    • 2
  • Francesca Abate
    • 1
  • Claudia Ferrari
    • 1
  • Emanuela Balbis
    • 2
  • Nicola Traverso
    • 2
  • Damiano Cottalasso
    • 2
  1. 1.Dietetics and Nutritional UnitSan Martino University HospitalGenoaItaly
  2. 2.Pathology InstituteGenoa UniversityGenoaItaly

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