动物营养学报 >

2017 , Vol. 29 >Issue 9: 3366 - 3374

DOI: https://doi.org/10.3969/j.issn.1006-267x.2017.09.042

研究简报 Short communications

赖氨酸对奶牛乳腺上皮细胞内乳脂肪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
陈璐(1990-),女,山西襄汾人,硕士研究生,从事奶牛营养研究。E-mail:1510560671@qq.com

收稿日期: 2017-02-24

  网络出版日期: 2017-09-08

基金资助

国家奶业"973计划"项目（2011CB1008003）

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Effects of Lysine on Expressions of Genes and Proteins Involved in 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: 2017-02-24

  Online published: 2017-09-08

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摘要

本试验旨在研究赖氨酸（Lys）对奶牛乳腺上皮细胞（BMECs）内乳脂肪合成相关基因和蛋白表达的影响，探讨Lys影响乳脂肪合成的机理。将第3代BMECs随机分为6组，每组6个重复，每个重复1个培养孔。各组培养基中Lys的浓度分别为0.5（基础培养基，对照）、1.0、2.0、4.0、8.0和16.0 mmol/L，37℃、5% CO2培养48 h后测定BMECs甘油三酯（TAG）含量、乳脂肪合成相关基因和蛋白的表达量。结果表明：BMECs内TAG含量（P=0.013）以及脂肪酸结合蛋白3（FABP3，P=0.001）、脂蛋白脂酶（LPL，P=0.096）、脂肪酸合成酶（FASN，P=0.003）、乙酰甘油磷酸脂酰转移酶6（AGPAT6，P=0.038）和甘油-3-磷酸酰基转移酶（GPAM，P=0.022）基因表达量对Lys呈显著或趋于显著的浓度依赖效应。FABP3基因表达量以2.0、4.0、8.0、16.0 mmol/L组和LPL基因表达量以1.0、2.0、4.0、8.0、16.0 mmol/L组显著高于0.5 mmol/L组（P<0.05）；FASN基因表达量以2.0 mmol/L组最高，显著高于16.0 mmol/L组（P<0.05）；硬脂酰辅酶A去饱和酶1（SCD1）基因表达量以2.0、4.0 mmol/L组显著高于其他组（P<0.05）；磷脂酸磷酸酯酶1（LPIN1）、嗜乳脂蛋白亚家族1成员1（BTN1A1）和黄嘌呤脱氢酶（XDH）基因表达量均以1.0、2.0、4.0、8.0 mmol/L组显著高于0.5 mmol/L组（P<0.05）；过氧化物酶体增殖物激活受体γ（PPARγ）基因及蛋白表达量均以2.0、4.0 mmol/L组显著高于0.5和8.0、16.0 mmol/L组（P<0.05）；固醇调节元件结合蛋白1（SREBP1）基因表达量以1.0、2.0、4.0 mmol/L组显著高于其他组（P<0.05），蛋白表达量以1.0 mmol/L组显著高于其他组（P<0.05）。但高浓度Lys抑制AGPAT6和GPAM的基因表达，AGPAT6基因表达量以2.0、4.0、8.0、16.0 mmol/L组显著低于0.5、1.0 mmol/L组（P<0.05），GPAM基因表达量以16.0 mmol/L组显著低于0.5、1.0、2.0、4.0 mmol/L组（P<0.05）。可见，Lys对BMECs的乳脂肪合成具有显著的促进效果，但高浓度的Lys抑制了乳脂肪合成相关基因的表达。本试验条件下，培养基中Lys适宜浓度为2.0~4.0 mmol/L。

关键词： 奶牛; 乳腺上皮细胞; 赖氨酸; 乳脂肪

本文引用格式

陈璐, 赵艳丽, 郭晓宇, 史彬林, 闫素梅 . 赖氨酸对奶牛乳腺上皮细胞内乳脂肪合成相关基因和蛋白表达的影响[J]. 动物营养学报, 2017 , 29(9) : 3366 -3374 . DOI: 10.3969/j.issn.1006-267x.2017.09.042

Abstract

This study was to detect the effects of lysine (Lys) on expressions of genes and proteins involved in milk fat synthesis in bovine mammary epithelial cells (BMECs) and discuss the mechanism of Lys regulating milk fat synthesis. The 3rd passage BMECs were divided into six groups with six replicates per group and one pore per replicate. Cells were cultured in medium containing 0.5 (basal medium, control), 1.0, 2.0, 4.0, 8.0 and 16.0 mmol/L Lys, respectively. The triglyceride (TAG) content, expressions of genes and proteins involved in milk fat synthesis were detected after 48 h cultivation at 37℃ and 5% CO2. The results showed as follows:TAG content (P=0.013) and gene expressions of fatty acid-binding protein 3 (FABP3, P=0.001), lipoprotein lipase (LPL, P=0.096), fatty acid synthase (FASN, P=0.003), 1-acylglycerol-3-phosphate O-acyltransferase 6 (AGPAT6, P=0.038) and glycerol-3-phosphate acyltrandferase (GPAM, P=0.022) acted dose-dependent on Lys at significant level or significant tendency. Compared with 0.5 mmol/L group, 2.0, 4.0, 8.0 and 16.0 mmol/L groups significantly increased gene expression of FABP3 (P<0.05), and 1.0, 2.0, 4.0, 8.0 and 16.0 mmol/L groups significantly increased gene expression of LPL (P<0.05). Gene expression of FASN in 2.0 mmol/L group was the highest, which was significantly higher than that in 16.0 mmol/L group (P<0.05). Gene expression of stearoyl-CoA desaturase 1 (SCD1) in 2.0 to 4.0 mmol/L groups was significantly higher than that in other groups (P<0.05). Gene expressions of phosphatidic acid phosphatase 1 (LPIN1), butyrophilin subfamily 1 member A1 (BTN1A1) and xanthine dehydrogenase (XDH) in 1.0, 2.0, 4.0 and 8.0 mmol/L groups were significantly higher than those in 0.5 mmol/L group. Compared with the 0.5, 8.0 and 16.0 mmol/L groups, 2.0 and 4.0 mmol/L groups significantly increased gene and protein expression of peroxisome proliferator-activated receptor-γ (PPARγ) (P<0.05). Gene expression of sterol regulatory element binding protein 1 (SREBP1) in 1.0, 2.0 and 4.0 mmol/L groups was significantly higher than that in the other groups (P<0.05), and protein expression of SREBP1 in 1.0 mmol/L group was significantly higher than that in the other groups (P<0.05). High dose of Lys decreased gene expressions of AGPAT6 and GPAM. Compared with 0.5 and 1.0 mmol/L groups, 2.0, 4.0, 8.0 and 16.0 mmol/L groups significantly decreased gene expression of AGPAT6 (P<0.05). Compared with 0.5, 1.0, 2.0 and 4.0 mmol/L groups, 16.0 mmol/L groups significantly decreased gene expression of GPAM (P<0.05). In conclusion, Lys significantly promote milk fat synthesis in BMECs, but high dose of Lys inhibits gene expressions related in milk fat synthesis. Under the conditions in the present study, 2.0 to 4.0 mmol/L of Lys is an optimal level in culture medium.

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

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