The Strong Antioxidant Sheep/Goat Whey Protein Protects Against mTOR Overactivation in Rats: A Mode of Action Mimicking Fasting

Whey protein, a by-product of the cheese industry, can be putatively used as a functional food due to its beneficial health properties. The main objective of the present study was to assess in vivo the effect of a sheep/goat whey protein on the plasma amino acid profile and mammalian target of rapamycin (mTOR), a regulator of skeletal myogenesis. A control group was fed with a standard commercial diet while the experimental group received a standard commercial diet plus sheep/goat whey protein for 28 days. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was conducted to determine plasma amino acid levels while the expression of p70-S6 Kinase 1 (p70-S6K1) in liver and quadriceps muscles was quantified and used as a biomarker of mTOR activity. The results obtained showed a decrease in the levels of essential and branched-chain amino acids (BCAAs) in the experimental group. Furthermore, p70-S6K1 expression was decreased in the liver of rats consumed whey protein. In conclusion, the reduction of amino acid levels and the concomitant inactivation of mTOR imply that whey could potentially act protectively against disorders induced by mTOR overactivation. Intriguingly, this mode of action mimics fasting, an approach with established advantageous health effects.

[1]  M. De Marchi,et al.  Short communication: Phenotypic characterization of total antioxidant activity of buffalo, goat, and sheep milk. , 2018, Journal of dairy science.

[2]  Xi Ma,et al.  Branched Chain Amino Acids: Beyond Nutrition Metabolism , 2018, International journal of molecular sciences.

[3]  D. Spandidos,et al.  Effects of sheep/goat whey protein dietary supplementation on the redox status of rats , 2018, Molecular medicine reports.

[4]  F. Mariotti Vegetarian and Plant-Based Diets in Health and Disease Prevention , 2017 .

[5]  M. De Marchi,et al.  Phenotypic characterisation of major mineral composition predicted by mid-infrared spectroscopy in cow milk , 2017 .

[6]  Arch G Mainous,et al.  Flipping the Metabolic Switch: Understanding and Applying Health Benefits of Fasting , 2017, Obesity.

[7]  T. Fenton,et al.  Protein intakes are associated with reduced length of stay: a comparison between Enhanced Recovery After Surgery (ERAS) and conventional care after elective colorectal surgery. , 2017, The American journal of clinical nutrition.

[8]  D. Kouretas,et al.  Increase in antioxidant activity by sheep/goat whey protein through nuclear factor-like 2 (Nrf2) is cell type dependent. , 2016, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[9]  Mee-Sup Yoon The Emerging Role of Branched-Chain Amino Acids in Insulin Resistance and Metabolism , 2016, Nutrients.

[10]  S. Groshen,et al.  Safety and feasibility of fasting in combination with platinum-based chemotherapy , 2016, BMC Cancer.

[11]  K. Yamauchi,et al.  Supplementation of protein-free diet with whey protein hydrolysates prevents skeletal muscle mass loss in rats , 2016 .

[12]  D. Kouretas,et al.  Antioxidant Effects of Sheep Whey Protein on Endothelial Cells , 2016, Oxidative medicine and cellular longevity.

[13]  M. De Marchi,et al.  Short communication: Selecting the most informative mid-infrared spectra wavenumbers to improve the accuracy of prediction models for detailed milk protein content. , 2016, Journal of dairy science.

[14]  C. Champagne,et al.  Plant protein and animal proteins: do they differentially affect cardiovascular disease risk? , 2015, Advances in nutrition.

[15]  M. Marchi,et al.  MILK COAGULATION PROPERTIES OF CATTLE BREEDS REARED IN ALPINE AREA , 2015 .

[16]  G. Theodoridis,et al.  Development and validation of a HILIC‐MS/MS multitargeted method for metabolomics applications , 2015, Electrophoresis.

[17]  Katsuhisa Horimoto,et al.  Plasma Free Amino Acid Profiles Predict Four-Year Risk of Developing Diabetes, Metabolic Syndrome, Dyslipidemia, and Hypertension in Japanese Population , 2015, Scientific Reports.

[18]  S. Groshen,et al.  A Periodic Diet that Mimics Fasting Promotes Multi-System Regeneration, Enhanced Cognitive Performance, and Healthspan. , 2015, Cell metabolism.

[19]  T. García-Gasca,et al.  Total phenolic compounds in milk from different species. Design of an extraction technique for quantification using the Folin-Ciocalteu method. , 2015, Food chemistry.

[20]  C. Lynch,et al.  Branched-chain amino acids in metabolic signalling and insulin resistance , 2014, Nature Reviews Endocrinology.

[21]  T. Unterman,et al.  Intermittent fasting vs daily calorie restriction for type 2 diabetes prevention: a review of human findings. , 2014, Translational research : the journal of laboratory and clinical medicine.

[22]  A. Tsatsakis,et al.  Antioxidant effects of whey protein on muscle C2C12 cells. , 2014, Food chemistry.

[23]  A. Jamurtas,et al.  Anti-inflammatory effects of a special carbohydrate-whey protein cake after exhaustive cycling in humans. , 2013, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[24]  Thomas J. Wang,et al.  A diabetes-predictive amino acid score and future cardiovascular disease. , 2013, European heart journal.

[25]  B. Caramelli,et al.  Review articleEffects of intermittent fasting on metabolism in menEfeitos do jejum intermitente no metabolismo humano , 2013 .

[26]  Miki Ebisuya,et al.  A fasting-responsive signaling pathway that extends life span in C. elegans. , 2013, Cell reports.

[27]  Shannon M. Conley,et al.  Retinal angiogenesis in the Ins2(Akita) mouse model of diabetic retinopathy. , 2013, Investigative ophthalmology & visual science.

[28]  D. Sabatini,et al.  Ragulator Is a GEF for the Rag GTPases that Signal Amino Acid Levels to mTORC1 , 2012, Cell.

[29]  A. Jamurtas,et al.  Effect of a special carbohydrate-protein cake on oxidative stress markers after exhaustive cycling in humans. , 2012, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[30]  Tuija Tammelin,et al.  Metabolic Signatures of Insulin Resistance in 7,098 Young Adults , 2012, Diabetes.

[31]  D. Sabatini,et al.  mTOR Signaling in Growth Control and Disease , 2012, Cell.

[32]  Vito Pistoia,et al.  Fasting Cycles Retard Growth of Tumors and Sensitize a Range of Cancer Cell Types to Chemotherapy , 2012, Science Translational Medicine.

[33]  J. Barłowska,et al.  Nutritional Value and Technological Suitability of Milk from Various Animal Species Used for Dairy Production , 2011 .

[34]  S. Adams Emerging perspectives on essential amino acid metabolism in obesity and the insulin-resistant state. , 2011, Advances in nutrition.

[35]  Aleksey A. Porollo,et al.  p62 is a key regulator of nutrient sensing in the mTORC1 pathway. , 2011, Molecular cell.

[36]  S. Gygi,et al.  Phosphoproteomic Analysis Identifies Grb10 as an mTORC1 Substrate That Negatively Regulates Insulin Signaling , 2011, Science.

[37]  D. Sabatini,et al.  The mTOR-Regulated Phosphoproteome Reveals a Mechanism of mTORC1-Mediated Inhibition of Growth Factor Signaling , 2011, Science.

[38]  V. Mootha,et al.  Metabolite profiles and the risk of developing diabetes , 2011, Nature Medicine.

[39]  B. Manning,et al.  mTOR couples cellular nutrient sensing to organismal metabolic homeostasis , 2011, Trends in Endocrinology & Metabolism.

[40]  Yibin Wang,et al.  Catabolism of Branched-Chain Amino Acids in Heart Failure: Insights from Genetic Models , 2011, Pediatric Cardiology.

[41]  D. Sabatini,et al.  mTOR: from growth signal integration to cancer, diabetes and ageing , 2010, Nature Reviews Molecular Cell Biology.

[42]  J. Geleijnse,et al.  Dietary Protein and Blood Pressure: A Systematic Review , 2010, PloS one.

[43]  D. Sabatini,et al.  Ragulator-Rag Complex Targets mTORC1 to the Lysosomal Surface and Is Necessary for Its Activation by Amino Acids , 2010, Cell.

[44]  M. Doreau,et al.  Simultaneous quantification of carotenoids, retinol, and tocopherols in forages, bovine plasma, and milk: validation of a novel UPLC method , 2010, Analytical and bioanalytical chemistry.

[45]  L. Cavacini,et al.  Structure and function of immunoglobulins. , 2010, The Journal of allergy and clinical immunology.

[46]  E. Tai,et al.  Insulin resistance is associated with a metabolic profile of altered protein metabolism in Chinese and Asian-Indian men , 2010, Diabetologia.

[47]  K. Varady,et al.  Short-term modified alternate-day fasting: a novel dietary strategy for weight loss and cardioprotection in obese adults. , 2009, The American journal of clinical nutrition.

[48]  Chin-Lee Wu,et al.  Insulin Stimulates Adipogenesis through the Akt-TSC2-mTORC1 Pathway , 2009, PloS one.

[49]  M. Diaz-Meco,et al.  p62 at the Crossroads of Autophagy, Apoptosis, and Cancer , 2009, Cell.

[50]  J. Blenis,et al.  Molecular mechanisms of mTOR-mediated translational control , 2009, Nature Reviews Molecular Cell Biology.

[51]  Svati H Shah,et al.  A branched-chain amino acid-related metabolic signature that differentiates obese and lean humans and contributes to insulin resistance. , 2009, Cell metabolism.

[52]  J. Auwerx,et al.  Adipose-specific knockout of raptor results in lean mice with enhanced mitochondrial respiration. , 2008, Cell metabolism.

[53]  T. P. Neufeld,et al.  Regulation of TORC1 by Rag GTPases in nutrient response , 2008, Nature Cell Biology.

[54]  David M. Sabatini,et al.  The Rag GTPases Bind Raptor and Mediate Amino Acid Signaling to mTORC1 , 2008, Science.

[55]  Matt Kaeberlein,et al.  Yeast Life Span Extension by Depletion of 60S Ribosomal Subunits Is Mediated by Gcn4 , 2008, Cell.

[56]  T. Vellai,et al.  Longevity pathways converge on autophagy genes to regulate life span in Caenorhabditis elegans , 2008, Autophagy.

[57]  M. Driscoll,et al.  A Role for Autophagy in the Extension of Lifespan by Dietary Restriction in C. elegans , 2008, PLoS genetics.

[58]  Pengxiang She,et al.  Obesity-related elevations in plasma leucine are associated with alterations in enzymes involved in branched-chain amino acid metabolism. , 2007, American journal of physiology. Endocrinology and metabolism.

[59]  J. Menéndez,et al.  Fatty acid synthase and the lipogenic phenotype in cancer pathogenesis , 2007, Nature Reviews Cancer.

[60]  A. Høstmark,et al.  Bovine milk in human nutrition – a review , 2007 .

[61]  K. Guan,et al.  Expanding mTOR signaling , 2007, Cell Research.

[62]  C. López-Otín,et al.  Tissue-specific Autophagy Alterations and Increased Tumorigenesis in Mice Deficient in Atg4C/Autophagin-3* , 2007, Journal of Biological Chemistry.

[63]  Nektarios Tavernarakis,et al.  eIF4E function in somatic cells modulates ageing in Caenorhabditis elegans , 2007, Nature.

[64]  Seung-Jae V. Lee,et al.  Lifespan extension by conditions that inhibit translation in Caenorhabditis elegans , 2007, Aging cell.

[65]  P. Kapahi,et al.  Inhibition of mRNA translation extends lifespan in Caenorhabditis elegans , 2007, Aging cell.

[66]  A. Pihlanto Antioxidative peptides derived from milk proteins , 2006 .

[67]  Yasushi Noguchi,et al.  Network analysis of plasma and tissue amino acids and the generation of an amino index for potential diagnostic use. , 2006, The American journal of clinical nutrition.

[68]  A. Roussel,et al.  Mitochondrial production of reactive oxygen species and incidence of age-associated lymphoma in OF1 mice: Effect of alternate-day fasting , 2005, Mechanisms of Ageing and Development.

[69]  F. Gaucheron The minerals of milk. , 2005, Reproduction, nutrition, development.

[70]  J. Bruce German,et al.  Milk beyond essential nutrients: The metabolic food , 2005 .

[71]  J. Cerhan,et al.  Associations of dietary protein with disease and mortality in a prospective study of postmenopausal women. , 2005, American journal of epidemiology.

[72]  N. Sonenberg,et al.  Upstream and downstream of mTOR. , 2004, Genes & development.

[73]  P. Garlick The nature of human hazards associated with excessive intake of amino acids. , 2004, The Journal of nutrition.

[74]  M. Zemel,et al.  Role of calcium and dairy products in energy partitioning and weight management. , 2004, The American journal of clinical nutrition.

[75]  M. Matsui,et al.  In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. , 2003, Molecular biology of the cell.

[76]  Govind Bhagat,et al.  Promotion of tumorigenesis by heterozygous disruption of the beclin 1 autophagy gene. , 2003, The Journal of clinical investigation.

[77]  Arnold J. Levine,et al.  Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[78]  B. Edgar,et al.  Rheb promotes cell growth as a component of the insulin/TOR signalling network , 2003, Nature Cell Biology.

[79]  E. Hafen,et al.  Rheb is an essential regulator of S6K in controlling cell growth in Drosophila , 2003, Nature Cell Biology.

[80]  J. Crespo,et al.  Two TOR complexes, only one of which is rapamycin sensitive, have distinct roles in cell growth control. , 2002, Molecular cell.

[81]  J. German,et al.  Whey Components: Millennia of Evolution Create Functionalities for Mammalian Nutrition: What We Know and What We May Be Overlooking , 2002, Critical reviews in food science and nutrition.

[82]  J. Blenis,et al.  Mammalian cell size is controlled by mTOR and its downstream targets S6K1 and 4EBP1/eIF4E. , 2002, Genes & development.

[83]  C. Metges,et al.  Whole-body nitrogen and splanchnic amino acid metabolism differ in rats fed mixed diets containing casein or its corresponding amino acid mixture. , 2001, The Journal of nutrition.

[84]  Tobias Schmelzle,et al.  TOR, a Central Controller of Cell Growth , 2000, Cell.

[85]  C. Gaudichon,et al.  Nutritional and Physiological Criteria in the Assessment of Milk Protein Quality for Humans , 2000, Journal of the American College of Nutrition.

[86]  A. Harper,et al.  Activation of liver branched-chain alpha-keto acid dehydrogenase in rats by excesses of dietary amino acids. , 1985, The Journal of nutrition.

[87]  S. Hauschildt,et al.  Influence of dietary nitrogen intake on mammalian branched chain alpha-keto acid dehydrogenase activity. , 1981, The Journal of nutrition.

[88]  S. Hauschildt,et al.  Effects of Branched-Chain α-Keto Acids on Enzymes Involved in Branched-Chain α-Keto Acid Metabolism in Rat Tissues , 1980 .

[89]  R. Wohlhueter,et al.  Coinduction of Rat Liver Branched Chain α-Keto Acid Dehydrogenase Activities , 1970 .

[90]  E. M. Brown,et al.  Nomenclature of the proteins of cows' milk--sixth revision. , 1965, Journal of dairy science.

[91]  T. S. P. S.,et al.  GROWTH , 1924, Nature.

[92]  H. Daniel,et al.  Branched-chain amino acids as biomarkers in diabetes. , 2016, Current opinion in clinical nutrition and metabolic care.

[93]  R. Tsutsumi,et al.  Peptides and Proteins in Whey and Their Benefits for Human Health , 2014 .

[94]  M. Hansen,et al.  A Role for Autophagy in the Extension of Lifespan by Dietary Restriction in -1 , 2011 .

[95]  Corby K. Martin,et al.  Alternate-day fasting in nonobese subjects: effects on body weight, body composition, and energy metabolism. , 2005, The American journal of clinical nutrition.

[96]  Lubert Stryer,et al.  Protein structure and function , 2005, Experientia.

[97]  A. Pihlanto,et al.  Bioactive peptides and proteins. , 2003, Advances in food and nutrition research.

[98]  S. Hauschildt,et al.  Effects of branched-chain alpha-keto acids on enzymes involved in branched-chain alpha-keto acid metabolism in rat tissues. , 1980, The Journal of nutrition.

[99]  R. Wohlhueter,et al.  Coinduction of rat liver branched chain alpha-keto acid dehydrogenase activities. , 1970, The Journal of biological chemistry.

[100]  Billy R. Hammond,et al.  [Foods]. , 1953, El Dia medico.