The effect of nanoemulsified methionine and cysteine on the in vitro expression of casein in bovine mammary epithelial cells
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Objective: Dairy cattle nutrient requirement systems acknowledge amino acid (AAs) requirements in aggregate as metabolizable protein (MP) and assume fixed efficiencies of MP used for milk protein. Regulation of mammary protein synthesis may be associated with AA input and milk protein output. The aim of this study was to evaluate the effect of nanoemulsified methionine and cysteine on the in-vitro expression of milk protein (casein) in bovine mammary epithelial cells (MAC-T cells). Methods: Methionine and cysteine were nonionized using Lipoid S 75 by high-speed homogenizer. The nanoemulsified AA particle size and polydispersity index were determined by dynamic light scattering correlation spectroscopy using a high-performance particle sizer instrument. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the cytotoxicity effect of AAs with and without nanoionization at various concentrations (100 to 500 μg/mL) in mammary epithelial cells. MAC-T cells were subjected to 100% of free AA and nanoemulsified AA concentration in Dulbecco's modified Eagle medium/nutrient mixture F-12 (DMEM/F12) for the analysis of milk protein (casein) expression by the quantitative reverse transcription polymerase chain reaction method. Results: The AA-treated cells showed that cell viability tended to decrease (80%) in proportion to the concentration before nanogenesis, but cell viability increased as much as 90% after nanogenesis. The analysis of the expression of genetic markers related to milk protein indicated that; αs2-casein increased 2-fold, κ-casein increased 5-fold, and the amount of unchanged β-casein expression was nearly doubled in the nanoemulsified methionine-treated group when compared with the free-nanoemulsified methionine-supplemented group. On the contrary, the non-emulsified cysteine-administered group showed higher expression of genetic markers related to milk protein αs2-casein, κ-casein, and β-casein, but all the genetic markers related to milk protein decreased significantly after nanoemulsification. Conclusion: Detailed knowledge of factors, such nanogenesis of methionine, associated with increasing cysteine and decreasing production of genetic markers related to milk protein (casein) will help guide future recommendations to producers for maximizing milk yield with a high level of milk protein casein.
Abstract
Objective: Dairy cattle nutrient requirement systems acknowledge amino acid (AAs) requirements in aggregate as metabolizable protein (MP) and assume fixed efficiencies of MP used for milk protein. Regulation of mammary protein synthesis may be associated with AA input and milk protein output. The aim of this study was to evaluate the effect of nanoemulsified methionine and cysteine on the in-vitro expression of milk protein (casein) in bovine mammary epithelial cells (MAC-T cells). Methods: Methionine and cysteine were nonionized using Lipoid S 75 by high-speed homogenizer. The nanoemulsified AA particle size and polydispersity index were determined by dynamic light scattering correlation spectroscopy using a high-performance particle sizer instrument. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to determine the cytotoxicity effect of AAs with and without nanoionization at various concentrations (100 to 500 μg/mL) in mammary epithelial cells. MAC-T cells were subjected to 100% of free AA and nanoemulsified AA concentration in Dulbecco's modified Eagle medium/nutrient mixture F-12 (DMEM/F12) for the analysis of milk protein (casein) expression by the quantitative reverse transcription polymerase chain reaction method. Results: The AA-treated cells showed that cell viability tended to decrease (80%) in proportion to the concentration before nanogenesis, but cell viability increased as much as 90% after nanogenesis. The analysis of the expression of genetic markers related to milk protein indicated that; αs2-casein increased 2-fold, κ-casein increased 5-fold, and the amount of unchanged β-casein expression was nearly doubled in the nanoemulsified methionine-treated group when compared with the free-nanoemulsified methionine-supplemented group. On the contrary, the non-emulsified cysteine-administered group showed higher expression of genetic markers related to milk protein αs2-casein, κ-casein, and β-casein, but all the genetic markers related to milk protein decreased significantly after nanoemulsification. Conclusion: Detailed knowledge of factors, such nanogenesis of methionine, associated with increasing cysteine and decreasing production of genetic markers related to milk protein (casein) will help guide future recommendations to producers for maximizing milk yield with a high level of milk protein casein.
참고문헌 (27)
- Arriola Apelo SI, "Current representation and future trends of predicting amino acid utilization in the lactating dairy cow", J Dairy Sci, 97, pp.4000~4017 (2014)
- Bionaz M, "Milk protein. Chapter 11", pp.285~324 (2012), InTech
- Hristov AN, "A meta-analysis examining the relationship among dietary factors, dry matter intake, and milk and milk protein yield in dairy cows", J Dairy Sci, 87, pp.2184~2196 (2004)
- Kim DH, "mTOR interacts with raptor to form a nutrient-sensitive complex that signals to the cell growth machinery", Cell, 110, pp.163~175 (2002)
- Wang C, "Effects of dietary supplementation of methionine and lysine on milk production and nitrogen utilization in dairy cows", J Dairy Sci, 93, pp.3661~3670 (2010)
- Appuhamy JA, "Isoleucine and leucine independently regulate mTOR signaling and protein synthesis in MAC-T cells and bovine mammary tissue slices", J Nutr, 142, pp.484~491 (2012)
- Hanigan MD, "Modelling mammary metabolism in the dairy cow to predict milk constituent yield, with emphasis on amino acid metabolism and milk protein production : model evaluation", J Theor Biol, 217, pp.311~330 (2002)
- Kimball SR, "Regulation of global and specific mRNA translation by amino acids", J Nutr, 132, pp.883~886 (2002)
- Moshel Y, "Role of amino acids in translational mechanisms governing milk protein synthesis in murine and ruminant mammary epithelial cells", J Cell Biochem, 98, pp.685~700 (2006)
- Yang Q, "TSC1", TSC2 and Rheb have different effects on TORC1 and TORC2 activity, 2006, pp.103, 6811~6816
- Bequette BJ, "Amino acid exchange by the mammary gland of lactating goats when histidine limits milk production", J Dairy Sci, 83, pp.765~775 (2000)
- Prizant RL, "Negative effects of the amino acids Lys, His, and Thr on S6K1 phosphorylation in mammary epithelial cells", J Cell Biochem, 105, pp.1038~1047 (2008)
- Burgos SA, "Nutrient availability and lactogenic hormones regulate mammary protein synthesis through the mammalian target of rapamycin signaling pathway", J Dairy Sci, 93, pp.153~161 (2010)
- Lopez A, "Bioactive packaging : turning foods into healthier foods through biomaterials", Trends Food Sci Technol, 17, pp.567~575 (2006)
- Schubert MA, "Characterization of surface-modified solid lipid nanoparticles(SLN) : Influence of lecithin and nonionic emulsifier", Eur J Pharm Biopharm, 61, pp.77~86 (2005)
- Erjavec V, "In vivo study of liposomes as drug carriers to oral mucosa using EPR oximetry", Int J Pharm, 307, pp.1~8 (2006)
- Kidd P, "A review of the bioavailability and clinical efficacy of milk thistle phytosome : a silybin-phosphatidylcholine complex(siliphos)", Altern Med Rev, 10, pp.193~203 (2005)
- Justin V, "Physicochemical characterization of a nanodispersion, obtained with a phase inversion temperature technique", pp.2015
- Basirico L, "Comparison between conjugated linoleic acid and essential fatty acids in preventing oxidative stress in bovine mammary epithelial cells", J Dairy Sci, 100, pp.2299~2309 (2017)
- Tagari H, "Mammary uptake, portal-drained visceral flux, and hepatic metabolism of free and peptide-bound amino acids in cows fed steam-flaked or dry-rolled sorghum grain diets", J Dairy Sci, 91, pp.679~697 (2008)[DOI]
- Backwell FRC, "Evidence for the utilization of peptides for milk protein synthesis in the lactating dairy goat in-vivo", Am J Physiol, 271, pp.R955~R960 (1996)
- Pan Y, "Methionine-containing peptides can be used as methionine sources for protein accretion in cultured C2C12 and MAC-T cells", J Nutr, 126, pp.232~241 (1996)
- Shennan DB, "Peptide aminonitrogen transport by the lactating rat mammary gland", Biochim Biophys Acta Biomem, 1373, pp.252~260 (1998)
- Yang JY, "Effects of methionine and methionylmethionine on expression of casein αs1 gene in cultured bovine mammary epithelial cells", J Agric Biotechnol, 15, pp.24~27 (2007)
- Groneberg DA, "Peptide transport in the mammary gland: expression and distribution of PEPT2 mRNA and protein", Am J Physiol Endocrinol Metab, 282, pp.E1172~E1179 (2002)
- Wu G, "Arginine metabolism and nutrition in growth, health and disease", Amino Acids, 37, pp.153~168 (2009)
- Raggio G, "Effect of level of metabolizable protein on splanchnic flux of amino acids in lactating dairy cows", J Dairy Sci, 87, pp.3461~3472 (2004)