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Effect of the ratios of unsaturated fatty acids on the expressions of genes related to fat and protein in the bovine mammary epithelial cells

  • R. Sheng
  • S. M. YanEmail author
  • L. Z. Qi
  • Y. L. Zhao
Article

Abstract

The objective of this study was to evaluate the effects of the different ratios of unsaturated fatty acids (UFAs) (oleic acid, linoleic acid, and linolenic acid) on the cell viability and triacylglycerol (TAG) content, as well as the mRNA expression of the genes related to lipid and protein synthesis in bovine mammary epithelial cells (BMECs). Primary cells were isolated from the mammary glands of Holstein dairy cows and were passaged twice. Afterward, the cells were randomly allocated to six treatments, five UFA-treated groups, and one control group. For all of the treatments, the the fetal bovine serum in the culture solution was replaced with fatty acid-free BSA (1 g/L), and the cells were treated with different ratios of oleic, linoleic, and linolenic acids (0.75:4:1, 1.5:10:1, 2:13.3:1, 3:20:1, and 4:26.7:1) for 48 h, which were group 1 to group 5. The control culture solution contained only fatty acid-free BSA without UFAs (0 μM). The results indicated that the cell viability was not affected by adding different ratios of UFAs, but the accumulation of TAG was significantly influenced by supplementing with different ratios of UFAs. Adding different ratios of UFAs suppressed the expression of ACACA and FASN but had the opposite effect on the abundances of FABP3 and CD36 mRNA. The expression levels of PPARG, SPEBF1, CSN1S1, and CSN3 mRNA in the BMECs were affected significantly after adding different ratios of UFAs. Our results suggested that groups 1, 2, and 3 (0.75:4:1, 1.5:10:1, and 2:13.3:1) had stronger auxo-action on fat synthesis in the BMECs, where group 3 (2:13.3:1) was the best, followed by group 4 (3:20:1). However, group 5 (4:26.7:1) was the worst. Genes related to protein synthesis in the BMECs were better promoted in groups 2 and 3, and group 3 had the strongest auxo-action, whereas the present study only partly examined the regulation of protein synthesis at the transcriptional level; more studies on translation level are needed in the future. Therefore, when combining fat and protein synthesis, group 3 could be obviously fat and protein synthesis in the BMECs concurrently. However, further studies are necessary to elucidate the mechanism for regulating fat and protein synthesis in the BMECs.

Keywords

Different ratios of UFAs Mammary epithelial cells mRNA Fat Protein 

Notes

Acknowledgments

The authors acknowledge the support of the national key basic research program of China (Project No. 2011CB100800). The authors are also grateful to Qi Lizhi, Zhao Yanli, and Jin Lu for their assistance during the experiments.

Reference

  1. Anderson SM, Rudolph MC, McManaman JL (2007) Key stages in mammary gland development. Secretory activation in the mammary gland: it’s not just about milk protein synthesis! Breast Cancer Res 9(1):204CrossRefPubMedCentralPubMedGoogle Scholar
  2. Bernard L, Leroux C, Chilliard Y (2008) Expression and nutritional regulation of lipogenic genes in the ruminant lactating mammary gland. Adv Exp Med Biol 606:67–108CrossRefPubMedGoogle Scholar
  3. Bionaz M, Loor J (2008a) ACSL1, AGPAT6, FABP3, LPIN1, and SLC27A6 are the most abundant isoforms in bovine mammary tissue and their expression is affected by stage of lactation. J Nutr 138:1019–1024PubMedGoogle Scholar
  4. Bionaz M, Loor J (2008b) Gene networks driving bovine milk fat synthesis during the lactation cycle. BMC Genomics 9:366CrossRefPubMedCentralPubMedGoogle Scholar
  5. Chen HY (2013) Effect of two different types of diets on fatty acid metabolism of dairy cows. Master Thesis, Gansu Agricultural University. 21–23Google Scholar
  6. Chilliard Y, Ferlay A, Mansbridge RM (2000) Ruminant milk fat plasticity: nutritional control of saturated, polyunsaturated, trans and conjugated fatty acids. Ann Zootech 49:181–205CrossRefGoogle Scholar
  7. Cui RL, Wang JQ (2012) Effects of 18-carbon fatty acids on cell proliferation and triacylglycerol accumulation in bovine mammary epithelial cells in vitro. Acta Veterinaria Et Zootechnica Sinica 43(7):1064–1070Google Scholar
  8. Desvergne B, Michalik L, Wahli W (2006) Transcriptional regulation of metabolism. Physiol Rev 86(2):465–514CrossRefPubMedGoogle Scholar
  9. Doege H, Stahl A (2006) Protein-mediated fatty acid uptake: novel insights from in vivo models. Physiology 21:259–268CrossRefPubMedGoogle Scholar
  10. Faergeman NJ, Wadum M, Feddersen S, Burton M, Kragelund BB (2007) Knudsen Acyl-CoA binding proteins; structural and functional conservation over 2000 MYA. Mol Cell Biochem 299(1–2):55–65CrossRefPubMedGoogle Scholar
  11. Harvatine KJ, Bauman DE (2006) SREBP1 and thyroid hormone responsive spot 14 (S14) are involved in the regulation of bovine mammary lipid synthesis during diet-induced milk fat depression and treatment with CLA. J Nutr 136(10):2468–2474PubMedGoogle Scholar
  12. Kadegowda AK, Piperova LS, Delmonte P (2008a) Abomasal infusion of butterfat increases milk fat in lactating dairy cows. J Dairy Sci 91:2370–2379CrossRefPubMedGoogle Scholar
  13. Kadegowda AK, Bionaz M, Piperova LS (2008b) Lipogenic gene expression in MAC-T cells is affected differently by fatty acids and enhanced by PPAR-gamma activation. J Dairy Sci 86(E-Suppl 2):566Google Scholar
  14. Keenan TW, Mather IH (2006) Intracellular origin of milk fat globules and the nature of the milk fat globule membrane. In Adv Dairy Chem Lip 2:137–171CrossRefGoogle Scholar
  15. Kim JE, Chen J (2004) Regulation of peroxisome proliferator-activated receptor-gamma activity by mammalian target of rapamycin and amino acids in adipogenesis. Diabetes 53:2748–2756CrossRefPubMedGoogle Scholar
  16. Laplante M, Sabatini DM (2009) mTOR signaling at a glance. J Cell Sci 122(20):3589–3594CrossRefPubMedCentralPubMedGoogle Scholar
  17. Lehner R, Kuksis A (1996) Biosynthesis of triacylglycerols. Progin Lipid Res 35:169–201CrossRefGoogle Scholar
  18. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using realtime quantitative PCR and the 2−ΔΔCT Method. Methods 25(4):402–408Google Scholar
  19. Ma XM, Blenis J (2009) Molecular mechanisms of mTOR mediated translational control. Nat Rev Mol Cell Biol 10:307–318CrossRefPubMedGoogle Scholar
  20. McFadden JW, Corl BA (2010) Activation of liver X receptor (LXR) enhances de novo fatty acid synthesis in bovine mammary epithelial cells. J Dairy Sci 93(10):4651–4658CrossRefPubMedGoogle Scholar
  21. Ma L, Corl BA (2012) Transcriptional regulation of lipid synthesis in bovine mammary epithelial cells by sterol regulatory element binding protein-1. J Dairy Sci 95:3743–3755CrossRefPubMedGoogle Scholar
  22. Mach N, Jacobs AAA, Kruijt L (2011) Alteration of gene expression in mammary gland tissue of dairy cows in response to dietary unsaturated fatty acids. Anim 5(8):1217–1230CrossRefGoogle Scholar
  23. Palmquist DL, Griinari JM (2006) Milk fatty acid composition in response to reciprocal combinations of sunflower and fish oils in the diet. Anim Feed Sci Tech 131:358–369Google Scholar
  24. Pauloin A, Chat S, Péchoux C (2010) Oleate and linoleate stimulate degradation of β-casein in prolactin-treated HC11 mouse mammary epithelial cells. Cell Tissue Res 340:91–102CrossRefPubMedGoogle Scholar
  25. Ramirez-Zacarias JL, Castro-Monozledo F, KuriHarcuch W (1992) Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with oil red O. Histochemistry 97:493–497Google Scholar
  26. Robenek H, Hofnagel O, Buers I, Lorkowski S, Schnoor M, Robenek MJ, Heid H, Troyer D, Severs NJ (2006) Butyrophilin controls milk fat globule secretion. Proc Natl Acad Sci U S A 103(27):10385–10390CrossRefPubMedCentralPubMedGoogle Scholar
  27. Soliman M, Kimura K, Ahmed M (2007) Inverse regulation of leptin mRNA expression by short- and long-chain fatty acids in cultured bovine adipocytes. Domest Anim Endocrinol 3:400–409CrossRefGoogle Scholar
  28. Soliman MM, Ahmed MM, Salah-eldin A (2011) Butyrate regulates leptin expression through different signaling pathways in adipocytes. J Veter Sci 12(4):319–323CrossRefGoogle Scholar
  29. Spitsberg VL, Elvina M, Ronald CG (1995). Association and Coexpression of Fatty-Acid-Binding Protein and Glycoprotein CD36 in the Bovine Mammary Gland. Eur J Biochem 230(3):872–878Google Scholar
  30. Sun XJ (2012) Effect and mechanism of eighteen carbon unsaturated fatty acid on mammary epithelial cells fat metabolism. Master Thesis, Inner Mongolia Agricultural University. 30–32Google Scholar
  31. Toshifumi A, Jeffrey MP, Nobuko I, Tamie N, Kenichi F, Takashi H, Frank JG (1998) Altered constitutive expression of fatty acid-metabolizing enzymes in mice lacking the peroxisome proliferator-activated receptor a (PPARa). J Bio Chem 273(10):5678–5684CrossRefGoogle Scholar
  32. Yang X, Yang C, Farberman A, Rideout TC, de Lange CF (2008) The mammalian target of rapamycin signaling pathway in regulating metabolism and growth. J Anim Sci 86(14):E35–E50Google Scholar
  33. Yonezawa T, Haga S, Kobayashi Y (2008a) Unsaturated fatty acids promote proliferation via ERK1/2 and Akt pathway in bovine mammary epithelial cells. Biochem Biophys Res Commun 367(4):729–735CrossRefPubMedGoogle Scholar
  34. Yonezawa T, Sanosaka M, Haga S (2008b) Regulation of uncoupling protein 2 expression by long-chain fatty acids and hormones in bovine mammary epithelial cells. Bioche Biophy Res Com 375(2):280–285CrossRefGoogle Scholar
  35. Yonezawa T, Yonekura S, Kobayashi Y (2004) Effects of long-chain fatty acids on cytosolic triacylglycerol accumulation and lipid droplet formation in primary cultured bovine mammary epithelial cells. J Dairy Sci 87:2527–2534CrossRefPubMedGoogle Scholar
  36. Zhao XW, Wang JQ, Sun P (2011) Effect of dietary supplementation with different fatty acid mixture on blood fatty acid composition and antioxidant capacity in dairy cows. J China Agric Univ 16:117–123Google Scholar

Copyright information

© The Society for In Vitro Biology 2015

Authors and Affiliations

  1. 1.College of Animal ScienceInner Mongolia Agricultural UniversityHuhhotChina

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