Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (23): 4892-4901.doi: 10.3864/j.issn.0578-1752.2011.23.016

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Effects of Exogenetic Trans-10, Cis-12 CLA on Genes Expression and Protein Synthesis of SREBP-1 Gene in Bovine Mammary Epithelial Cells

 WANG  Hong-Fang, LIU  Hong-Yun, YANG  Wei-Ren, LIU  Jian-Xin, YANG  Zai-Bin   

  1. 1.山东农业大学动物科技学院，山东泰安 271018
    2.浙江大学动物科技学院奶业科学研究所，杭州 310029
  • Received:2010-12-13 Online:2011-12-01 Published:2011-03-07

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Abstract: 【Objective】The purpose of the study is to explore the effects of exogenetic t10c12 CLA on genes expression and protein synthesis of SREBP-1 in bovine mammary epithelial cells (BMECs). 【Method】 BMECs used in experiment were achieved by tissue culture methods. Lipids in cytoplasm were stained using oil red. The total RNA was extracted from BMECs treated with different gradients of t10c12 CLA. And then the expression levels of various genes were quantified by quantitative reverse transcription PCR (QRT-PCR) analysis. Total proteins extracted with protein extraction kit were used in SDS-PAGE and western blotting. 【Result】Concentration of cytosolic triacylglycerols (TAG) decreased linearly (P=0.0003) with the increasing t10c12 CLA concentration. The addition of t10c12 CLA had no effects on the expression of SREBP-1 gene and other genes related to post-translation process of SREBP-1 and the protein synthesis of precursor SREBP-1(pSREBP-1), maybe, it inhibited the synthesis or promoted the degradation of bio-active SREBP-1(bSREBP-1).【Conclusion】T10c12 CLA didn’t affect the gene expression and protein translation of SREBP-1 and inhibited cytosolic TAG accumulation. According to the results, it was speculated that t10c12 CLA maybe have some effects on activation of pSREB-1 and degradation of bSREBP-1.

Key words: t10c12 CLA, bovine mammary epithelial cells, triacylglycerols, SREBP-1

[1]冯仰廉. 反刍动物营养学. 北京: 科学出版社, 2004: 418-421.

Feng Y L. Ruminant Nutrition. Beijing: Science Press, 2004: 418-421. (in Chinese)

[2]Bhattacharya A, Banu J, Rahman M, Causey J, Fernandes G. Biological effects of conjugated linoleic acids in health and disease. The Journal of Nutritional Biochemistry, 2006, 17: 789-810.

[3]Kelley N S, Hubbard N E, Erickson K L. Conjugated linoleic acid isomers and cancer. The Journal of Nutrition, 2007, 137: 2599-2607.

[4]Lee K N, Kritchevsky D, Pariza M W. Conjugated linoleic acid and atherosclerosis in rabbits. Atherosclerosis, 1994, 108: 19-25.

[5]O’Shea M, Bassaganya-Riera J, Mohede I C. Immunomodulatory properties of conjugated linoleic acid. The American Journal of Clinical Nutrition, 2004, 79: S1199- S1206.

[6]魏登邦, 张宝琛. 动物体内共轭亚油酸产生机制及影响含量的因素.青海大学学报, 2002, 20(5): 38-40.

Wei D B, Zhang B C. The formatting mechanism and the factors of influencing the contents of CLA in animals. Journal of Qinghai University, 2002, 20(5): 38-40. (in Chinese)

[7]Dhiman T R, Nam S H, Ure A L. Factors affecting conjugated linoleic acid content in milk and meat. Critical Reviews in Food Science and Nutrition, 2005, 45: 463-482.

[8]郑会超, 刘建新, 姚建红, 袁　谦, 吴跃明. 添加植物油对奶牛乳脂率及乳中共轭亚油酸含量的影响. 营养学报, 2005, 27(6): 523-524.

Zheng H C, Liu J X, Yao J H, Yuan Q, Wu Y M. Effects of dietary addition of vegetable oils on milk fat percentage of lactating cows and the cotents of conjugated linoleic acids in milk. Acta Nutrimenta Sinica, 2005, 27(6): 523-524. (in Chinese)

[9]尹福泉, 嘎尔迪, 刘瑞芳, 于  磊, 卢寿锋, 齐智利. 不同植物油对奶牛瘤胃液、血浆中脂肪酸及乳脂中共轭亚油酸组成的影响. 动物营养学报, 2008, 20(1): 63-68.

Yin F Q, Ga E D, Liu R F, Yu L, Lu S F, Qi Z L. Effects of different vegetable oils on fatty acids in ruminal fluid and blood plasma and milk cla of lactating cows. Chinese Journal of Animal Nutration, 2008, 20(1): 63-68. (in Chinese)

[10]尹福泉, 嘎尔迪, 刘瑞芳, 于  磊, 卢寿锋, 齐智利. 日粮中添加油料籽实对奶牛生产性能及乳脂脂肪酸组成的影响. 动物营养学报, 2008, 20(3): 261-267.

Yin F Q, Ga E D, Liu R F, Yu L, Lu S F, Qi Z L. Effects of dietary supplementation of different oil seeds on production performance and milk fatty acids of dairy cows. Chinese Journal of Animal Nutration, 2008, 20(3): 261-267. (in Chinese)

[11]陈艳珍, 梁成山, 张  鹰. 共扼亚油酸对奶牛产奶量和乳脂率的影响. 中国饲料, 2002, 18: 11-12.

Chen Y Z, Liang C S, Zhang Y. Effects of conjugated linoleic acid on milk yield and milk fat percentage of dairy cows. China Feed, 2002, 18: 11 -12. (in Chinese)

[12]Baumgard L H, Corl B A, Dwyer D A, Saebo A, Bauman D E. Identification of the conjugated linoleic acid isomer that inhibits milk fat synthesis. American Journal of Physiology Regulatory Integrative and Comparative Physiology, 2000, 278: R179-R184.

[13]Harvatine K J, Perfield J W, Bauman D E. Expression of enzymes and key regulators of lipid synthesis is upregulated in adipose tissue during CLA-induced milk fat depression in dairy cows. The Journal of Nutrition, 2009, 139(5): 849-854.

[14]Baumgard L H, Matitashvili E, Corl B A, Dwyer D A, Bauman D E. Trans-10, cis-12 conjugated linoleic acid decreases lipogenic rates and expression of genes involved in milk lipid synthesis in dairy cows. Journal of Dairy Science, 2002, 85: 2155-2163.

[15]Peterson D G, Baumgard L H, Bauman D E. Short communication: milk fat response to low doses of trans-10, cis-12 conjugated linoleic acid (CLA). Journal of Dairy Science, 2002, 85: 1764-1766.

[16]Peterson D G, Matitashvili E A, Bauman D E. Diet-induced milk fat depression in dairy cows results in increased trans-10, cis-12 CLA in milk fat and coordinate suppression of mRNA abundance for mammary enzymes involved in milk fat synthesis. The Journal of Nutrition, 2003, 133: 3098-3102.

[17]Edwards P A, Tabor D, Kast H R, Venkateswaran A. Regulation of gene expression by SREBP and SCAP. Biochimica et Biophysica Acta, 2000, 1529(1-3): 103-113.

[18]Peterson D G, Matitashvili E A, Bauman D E. The inhibitory effect of trans-10, cis-12 CLA on lipid synthesis in bovine mammary epithelial cells involves reduced proteolytic activation of the transcription factor SREBP-1. The Journal of Nutrition, 2004, 134: 2523-2527.

[19]Stoeckman A K, Towlet H C. The role of srebp-1c in nutritional regulation of lipogenic enzyme gene expression. The Journal of Biological Chemistry, 2002, 277(30): 27029-27035.

[20]Min Y, Fischer M, Deeg M A, Crabb D W. Ethanol induces fatty acid synthesis pathways by activation of sterol regulatory element-binding protein (SREBP). The Journal of Biological Chemistry, 2002, 277(32): 29342-29347.

[21]Ramirez-Zacarias J L, Castro-Muñozledo F, Kuri-Harcuch W. Quantitation of adipose conversion and triglycerides by staining intracytoplasmic lipids with oil red O. Histochemistry, 1992, 97: 493-497.

[22]Annison E F, Bicherstaffe R, Linzell J L. Glucose and fatty acid metabolism in cows producing milk of low fat content. The Journal of Agricultural Science(Cambridge), 1974, 82: 87-95.

[23]Ebner K E, Hoover C R, Hageman E C, Larson B L. Cultivation and properties of bovine mammary cell cultures. Experimental Cell Research, 1961, 23: 373-381.

[24]Thordarson G, Ogren L, Day J R, Bowens K, Fielder P, Talamantes F. Mammary gland development and alpha-lactalbum in production in hypophysectomized, pregnantmice. Biology Reproduction, 1989, 40(3): 517-525.

[25]Schmid E, Schiller D L, Grund C, Stadler J, Franke W W. Tissue type-specific expression of intermediate filament proteins in a cultured epithelial cell line from bovine mammary gland. The Journal of Cell Biology, 1983, 96(1): 37-50.

[26]Gibson C A, Vega J R, Baumrucker C R. Establishment and characterization of bovine mammary epithelial cell lines. In Vitro Cellular and Developmental Biology, 1991, 27A(7): 585-594.

[27]Huynh H T, Robitailte G, Turner J. Establishment of bovine mammary epithelial cells (MAC-T): an in vitro model for bovine lactation. Experimental Cell Research, 1991, 197(2): 191-199.

[28]Huynh H, Pollak M. HH2a, an immortalized bovine mammary epithelial cell line expresses the gene encoding mammary derived growth inhibitor (MDG I). In Vitro Cellular and Developmental Biology Animal, 1995, 31A: 25-29.

[29]Zavizion B, van Duffelen M, Schaeffer W, Politis I. Establishment and characterization of a bovine mammary epithelial cell line with unique properties. In Vitro Cellular and Developmental Biology Animal, 1996, 32: 138-148.

[30]Guzzman R C, Osborn R C, Richards J E, Nandi S. Effects of phorbolesters on normal and tumorous mouse mammary epithelial cells embedded in collagengels. Journal of the National Cancer Institute, 1983, 71: 69-73.

[31]Sørensen B M, Kazala E C, Murdoch G K, Keating A K, Cruz-Hernandez C, Wegner J, Kennelly J J, Okine E K, Weselake R J. Effect of CLA and other C18 unsaturated fatty acids on DGAT in bovine milk fat biosynthetic systems. Lipids, 2008, 43: 903-912.

[32]Zhao K, Liu H Y, Zhou M M, Liu J X. Establishment and characterization of lactating bovine mammary epithelial cell model for milk synthesis study. Cell Biology International, 2010, 34(7): 717-721.

[33]Miller W J, Tyznik W J, Allen N N, Sorensen D K. The treatment of ketosis in dairy cows by oral administration of sodium acetate. Journal of the American Veterinary Medical Association, 1954, 124(925): 291-294.

[34]Vallance W S, McClymont G L. Depression in percentage of milk fat by parenteral glucose infusion and glycerol feeding. Nature, 1959, 183(4659): 466-476.

[35]Jenny B F, Polan C E, Thye F W. Effects of high grain feeding and stage of lactation on serum insulin, glucose and milk fat percentage in lactating cows. The Journal of Nutrition, 1974, 104: 379-385.

[36]Davis C L, Brown R E. Availability and metabolism of various substrates in ruminants. iv. glucose metabolism in the young calf and growing steer. Journal of Dairy Science, 1962, 45(4): 513-516.

[37]Bauman D E, Griinari J M. Regulation and nutritional manipulation of milk fat: low-fat milk syndrome. Advances Experimental Medicine and Biology, 2000, 480: 209-216.

[38]Baumgard L H, Sangster J K, Bauman D E. Milk fat synthesis in dairy cows is progressively reduced by increasing supplemental amounts of trans-10, cis-12 conjugated linoleic acid(CLA). The Journal of Nutrition, 2001, 131: 1764-1769.

[39]Loor J J, Herbein J H. Reduced fatty acid synthesis and desaturation due to exogenous trans-10, cis-12 CLA in cows fed oleic or linoleic oil. Journal of Dairy Science, 2003, 86: 1354-1369.

[40]Hansen H O, Knudsen J. Effect of exogenous long-chain fatty acids on individual fatty acid synthesis by dispersed ruminant mammary gland cells. Journal of Dairy Science, 1987, 70(7): 1350-1364.

[41]Yonezawa T, Yonekura S, Kobayashi Y, Hagino A, Katoh K, Obara  Y. Effects of long-chain fatty acids on cytosolic triacylglycerol accumulation and lipid droplet formation in primary cultured bovine mammary epithelial cells. Journal of Dairy Science, 2004, 87: 2527-2537.

[42]Kadegowda A K G, Bionaz M, Piperova L S, Erdman R A, Loor J J. Peroxisome proliferator-activated receptor-γ activation and long-chain fatty acids alter lipogenic gene networks in bovine mammary epithelial cells to various extents. Journal of Dairy Science, 2009, 92(9): 4276-4289.

[43]Piperova L S, Teter B B, Bruckental I, Sampugna J, Mills S E, Yurawecz M P, Fritsche J, Ku K, Erdman R A. Mammary lipogenic enzyme activity, trans-fatty acids and conjugated linoleic acids are altered in lactating dairy cows fed a milk fat-depressing diet. The Journal of Nutrition, 2000, 130(10): 2568-2574.

[44]Harvatine K J, Perfield J W, Bauman D E. Expression of enzymes and key regulators of lipid synthesis is upregulated in adipose tissue during CLA-induced milk fat depression in dairy cows. The Journal of Nutrition, 2009, 139(5): 849-854.

[45]Bauman D E, Perfield J W, Harvatine K J, Baumgard L H. Regulation of fat synthesis by conjugated linoleic acid: lactation and the ruminant model. The Journal of Nutrition, 2008, 138(2): 403-409.

[46]Espenshade P J. SREBPs: sterol-regulated transcription factors. Journal of Cell Science, 2006, 119(6): 973-976.

[47]Botolin D, Wang Y, Christian B, Jump D B. Docosahexaneoic acid(22:6, n-3) regulates rat hepatocyte SREBP-1 nuclear abundance by Erk- and 26S proteasome-dependent pathways. Journal of Lipid Research, 2006, 47: 181-192.
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