动物营养学报 >

2017 , Vol. 29 >Issue 3: 961 - 969

DOI: https://doi.org/10.3969/j.issn.1006-267x.2017.03.029

研究简报 Short communications

蛋氨酸对奶牛乳腺上皮细胞内乳脂合成相关基因和蛋白表达的影响

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  • 内蒙古农业大学动物科学学院, 呼和浩特 010018
赵艳丽(1986-),女,陕西榆林人,博士研究生,从事奶牛乳腺上皮细胞内乳脂乳蛋白合成调控研究。E-mail:ylzhao2010@163.com

收稿日期: 2016-08-30

  网络出版日期: 2017-03-09

基金资助

国家奶业“973计划”项目（2011CB1008003）

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Effects of Methionine on Expression of Genes and Proteins Related with Milk Fat Synthesis in Bovine Mammary Epithelial Cells

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  • Collage of Animal Science, Inner Mongolia Agriculture University, Hohhot 010018, China

Received date: 2016-08-30

  Online published: 2017-03-09

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摘要

本试验旨在研究蛋氨酸（Met）对奶牛乳腺上皮细胞（BMECs）内乳脂合成相关基因和蛋白表达的影响，以探讨Met对乳脂合成的影响机理。将第3代BMECs随机分为6个处理（每个处理6个重复），培养液中Met浓度分别为0.13、0.26、0.39、0.52、0.65和0.78 mmol/L。在37℃、5% CO2条件下培养48 h后测定BMECs内甘油三酯（TG）的含量及乳脂合成相关基因和过氧化物酶体增殖物激活受体γ（PPARγ）与固醇调节元件结合蛋白1（SREBP1）蛋白的相对表达量。结果显示：Met浓度对BMECs内TG含量无显著影响（P>0.05）。0.52~0.78 mmol/L Met处理BMECs内脂肪酸结合蛋白3（FABP3）、乙酰辅酶A羧化酶A（ACACA）和PPARγ基因的相对表达量均显著高于其他处理（P<0.05）。BMECs内脂蛋白脂酶（LPL）基因的相对表达量以0.26~0.39 mmol/L Met处理较高，显著高于0.65~0.78 mmol/L Met处理（P<0.05）。脂肪酸合成酶（FASN）基因的相对表达量以0.39~0.52 mmol/L Met处理较高，且0.52 mmol/L Met处理显著高于0.13~0.26 mmol/L和0.65~0.78 mmol/L Met处理（P<0.05）。Met浓度可显著影响SREBP1和乙酰甘油磷酸脂酰转移酶6（AGPAT6）基因及SREBP1和PPARγ蛋白的表达（P<0.05），均以0.26 mmol/L Met处理的相对表达量最高。结果表明，Met浓度影响BMECs内脂肪酸摄取、从头合成的基因的表达及乳脂合成调控因子PPARγ和SREBP1的基因和蛋白的表达，Met浓度为0.26~0.52 mmol/L时对BMECs内脂肪酸从头合成及长链脂肪酸摄取的促进效果较好。

关键词： 奶牛; 乳腺上皮细胞; 蛋氨酸; 乳脂

本文引用格式

赵艳丽, 陈璐, 史彬林, 郭晓宇, 闫素梅 . 蛋氨酸对奶牛乳腺上皮细胞内乳脂合成相关基因和蛋白表达的影响[J]. 动物营养学报, 2017 , 29(3) : 961 -969 . DOI: 10.3969/j.issn.1006-267x.2017.03.029

Abstract

This experiment was to study the effects of methionine (Met) on expression of genes and proteins related with milk fat synthesis in bovine mammary epithelial cells (BMECs), in order to investigate the mechenism of Met regulating milk fat synthesis. The third generation of BMECs were divided into six treatments with six replicates per treatment, and cultured in culture mediums with 0.13, 0.26, 0.39, 0.52, 0.65 and 0.78 mmol/L Met, respectively. The triglyceride (TG) content, the relative expression levels of genes related with milk fat synthesis, as well as the protein relative expression levels of peroxisome proliferator-activated receptor-γ (PPARγ) and sterol regulatory element binding protein 1 (SREBP1) in BMECs after 48 h incubation at 37℃ and 5% CO2 were detected. The results showed that Met concentration had no significant effect on TG content in BMECs (P>0.05). The gene relative expression levels of fatty acid-binding protein 3 (FABP3), acetyl-CoA carboxylase A (ACACA) and PPARγ in BMECs of 0.52 to 0.78 mmol/L Met treatments were significantly higher than those of other treatments (P<0.05). The gene relative expression level of lipoprotein lipase (LPL) in BMECs of 0.26 to 0.39 mmol/L Met treatments was higher, which significantly higher than that of 0.65 to 0.78 mmol/L treatments (P<0.05). The gene relative expression level of fatty acid synthase (FASN) in BMECs of 0.39 to 0.52 mmol/L Met treatments was higher, and it of 0.52 mmol/L Met treatment was significantly higher than that of 0.13 to 0.26 mmol/L and 0.65 to 0.78 mmol/L Met treatments (P<0.05). Met concentration could significantly affect the gene expression of SREBP1 and 1-acylglycerol-3-phosphate O-acyltransferase 6 (AGPAT6) and the protein expression of SREBP1 and PPARγ in BMECs (P<0.05), and the highest relative expression levels of them were all observed in 0.26 mmol/L Met treatment. These results demonstrate that Met concentration affects the expression of genes related with long-chain fatty acids uptake and de novo synthesis of fatty acids, as well as the gene and protein expression of SREBP1 and PPARγ which were milk fat synthesis regulatory factors in BMECs. Met with the concentrations of 0.26 to 0.52 mmol/L can produce the better effects for long-chain fatty acids uptake and de novo synthesis of fatty acids in BMECs.

Key words： dairy cow; bovine mammary epithelial cells; methionine; milk fat

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