动物营养学报 >

2017 , Vol. 29 >Issue 11: 4143 - 4150

DOI: https://doi.org/10.3969/j.issn.1006-267x.2017.11.038

研究简报 Short Communications

脂肪酸对奶牛乳腺上皮细胞乳蛋白和乳脂合成相关基因表达量的影响

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  • 内蒙古农业大学动物科学学院, 呼和浩特 010018
张花(1991-),女,山西朔州人,硕士研究生,从事反刍动物营养生理及瘤胃微生态研究。E-mail:xiayeyuye@126.com

收稿日期: 2017-04-18

  网络出版日期: 2017-10-31

基金资助

国家自然科学基金（31360559）

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Effects of Fatty Acids on Expression Levels of Genes Involved in Milk Protein and Milk Fat Synthesis in Bovine Mammary Epithelial Cells

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  • College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China

Received date: 2017-04-18

  Online published: 2017-10-31

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摘要

本试验旨在探寻促进奶牛乳腺上皮细胞（BMECs）乳蛋白和乳脂合成的短链脂肪酸（乙酸、β-羟丁酸）和长链脂肪酸（油酸、亚油酸、亚麻酸）的组合添加模式，为调控乳成分合成提供理论依据。BMECs经分离、纯化后，选取第2代细胞，分为5组，对照组不添加脂肪酸，Ⅰ组和Ⅱ组添加的乙酸、β-羟丁酸浓度比例均为2.0（9.60 mmol/L）∶1.0（4.80 mmol/L），油酸、亚油酸、亚麻酸的浓度比例分别为2.0（17.30 μmol/L）∶13.3（115.05 μmol/L）∶1.0（8.65 μmol/L）和9.6（75.20 μmol/L）∶7.4（58.00 μmol/L）∶1.0（7.80 μmol/L）；Ⅲ组和Ⅳ组添加的乙酸、β-羟丁酸的浓度比例均为1.0（7.20 mmol/L）∶1.0（7.20 mmol/L），油酸、亚油酸、亚麻酸的浓度比例分别为2.0∶13.3∶1.0和9.6∶7.4∶1.0，各组添加的短链脂肪酸（SCFA）和长链脂肪酸（LCFA）总浓度为14.541 mmol/L，每组3个重复。培养24 h后，检测细胞相对增殖率（RGR）、甘油三酯（TAG）的合成量以及乳蛋白和乳脂合成相关基因的表达量。结果表明：1）试验组BMECs RGR及TAG的合成量均显著高于对照组（P<0.05）；Ⅰ组RGR最高，TAG合成量最多。2）与对照组相比，Ⅱ组显著提高了核糖体蛋白S6激酶1（S6K1）、κ-酪蛋白（CSN3）基因的表达量（P<0.05）；Ⅳ组显著提高了CSN3、蛋白激酶B （AKT）、S6K1、真核翻译起始因子4E结合蛋白1（4EBP1）基因的表达量（P<0.05）；试验组信号转导和转录激活因子5（STAT5）基因的表达量显著降低（P<0.05）。3）与对照组相比，试验组二酰甘油酰基转移酶2（DGAT2）基因的表达量显著提高（P<0.05），脂肪酸合成酶（FASN）基因的表达量显著降低（P<0.05）。综上所述，在培养液中添加7.20 mmol/L乙酸、7.20 mmol/L β-羟丁酸、75.20 μmol/L油酸、58.00 μmol/L亚油酸、7.80 μmol/L亚麻酸对BMECs乳蛋白和乳脂合成相关基因的表达量有较好的促进作用。

关键词： 奶牛乳腺上皮细胞; 短链脂肪酸; 长链脂肪酸; 乳蛋白; 乳脂

本文引用格式

张花, 李大彪, 邢媛媛, 孙梅 . 脂肪酸对奶牛乳腺上皮细胞乳蛋白和乳脂合成相关基因表达量的影响[J]. 动物营养学报, 2017 , 29(11) : 4143 -4150 . DOI: 10.3969/j.issn.1006-267x.2017.11.038

Abstract

The aim of this study was to explore combined supplemental model of short chain fatty acids (acetic acid, β-hydroxybutyric acid) and long chain fatty acids (oleic acid, linoleic acid, linolenic acid) to promote milk protein and milk fat synthesis in bovine mammary epithelial cells (BMECs), to provide theoretical basis for the regulation of synthesis of milk composition. BMECs were separated and purified, and the second generation of cells was selected and divided into 5 groups with 3 replicates per group. No fatty acid was added in control group; in groups Ⅰ and Ⅱ, the concentration radio of supplied acetate acid and β-hydroxybutyric acid was 2.0 (9.60 mmol/L):1.0 (4.80 mmol/L), however, the concentration radio of oleic acid, linoleic acid and linolenic acid was 2.0 (17.30 μmol/L):13.3 (115.05 μmol/L):1.0 (8.65 μmol/L) and 9.6 (75.20 μmol/L):7.4 (58.00 μmol/L):1.0 (7.80 μmol/L), respectively; in groups Ⅲ and Ⅳ, the concentration radio of acetate acid and β-hydroxybutyric acid was 1.0:1.0, however, the concentration radio of oleic acid, linoleic acid and linolenic acid was 2.0:13.3:1.0 and 9.6:7.4:1.0, respectively. The total concentration of short chain fatty acids and long chain fatty acids was 14.541 mmol/L. After cultured for 24 h, relative growth rate (RGR), triglyceride (TAG) synthesis amount and expression levels of genes involved in milk protein and milk fat synthesis were measured. The results showed as follows:1) RGR and TAG synthesis amount of BMECs in experimental groups were significantly higher as comparing with control group (P<0.05); RGR and TAG synthesis amount were the highest in group Ⅰ. 2) Compared with control group, expression levels of ribosomal protein S6kinase (S6K1) and κ-casein (CSN3) genes in group Ⅱ were significantly increased (P<0.05); expression levels of CSN3, protein kkinase B(AKT), S6K1 and eukaryotic initiation 4E binding protein (4EBP1) gene in group Ⅳ was significantly increased (P<0.05); expression level of signal transduction and transcriptional activation factor 5 (STAT5) gene in experimental groups was significantly decreased (P<0.05). 3) Compared with control group, expression level of diacylgycerol acyltransferase 2 (DGAT2) gene in experimental groups was significantly increased (P<0.05), and expression level of fatty acid synthase (FASN) gene was significantly decreased (P<0.05). In conclusion, expression levels of genes involved in milk protein and milk fat synthesis has a better effect when culture medium is added with 7.20 mmol/L acetate acid, 7.20 mmol/L β-hydroxybutyric acid, 75.20 μmol/L oleic acid, 58.00 μmol/L linoleic acid, and 7.80 μmol/L linolenic acid.

Key words： bovine mammary; short chain fatty acid; long chain fatty acid; milk protein; milk fat

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