动物营养学报 >

2015 , Vol. 27 >Issue 1: 196 - 203

DOI: https://doi.org/10.3969/j.issn.1006-267x.2015.01.024

饲料营养 Feed Science and Technology

β-羟丁酸对奶牛乳腺上皮细胞内乳脂肪合成及其相关基因相对表达量的影响

  • 常晨城 ,
  • 齐利枝 ,
  • 闫素梅 ,
  • 生冉 ,
  • 赵艳丽
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  • 内蒙古农业大学动物科学学院, 呼和浩特 010018

收稿日期: 2014-07-11

  网络出版日期: 2015-01-10

基金资助

奶业"973计划"(2011CB100800)

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Effects of β-Hydroxybutyric Acid on Relative Expression Levels of Genes Related to Milk Fat Synthesis in Bovine Mammary Epithelial Cells

  • CHANG Chencheng ,
  • QI Lizhi ,
  • YAN Sumei ,
  • SHENG Ran ,
  • ZHAO Yanli
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  • College of Animal Science, Inner Mongolia Agricultural University, Huhhot 010018, China

Received date: 2014-07-11

  Online published: 2015-01-10

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摘要

本试验主要研究了不同浓度的β-羟丁酸(BHBA)对奶牛乳腺上皮细胞(BMECs)活力、甘油三酯(TAG)含量、脂滴形成以及乳脂肪合成相关基因转录水平的影响。将传至第3代的BMECs悬液(1×105个/孔)接种于细胞培养板上,每孔加入含10%胎牛血清(FBS)的DMEM/F12培养液,于37 ℃的5%二氧化碳(CO2)培养箱培养48 h。再将培养48 h的BMECs随机分配到6个组,各组向培养孔中加入含不同浓度BHBA的DMEM/F12培养液,培养液中的FBS用1 g/L无脂肪酸的牛血清白蛋白(BSA)代替,并使反应体系中BHBA的最终浓度分别为0(对照)、0.58、1.16、2.32、4.64和9.28 mmol/L。置于37 ℃的5%CO2培养箱继续培养48 h。试验结果显示:随着BHBA浓度的增加,BMECs活力[(相对增殖率(RGR)]呈显著的二次曲线增加(P=0.041),其中BMECs活力以0.58~4.64 mmol/L BHBA组较高,9.28 mmol/L BHBA组较低;低浓度(0.58~2.32 mmol/L)的BHBA可促进BMECs内脂滴的形成,而较高浓度(4.64~9.28 mmol/L)的BHBA对脂滴形成的促进作用减弱;BHBA与TAG含量及乳脂肪合成相关基因脂肪酸合成酶(FASN)、乙酰辅酶A羧化酶α(ACACA)、硬脂酰辅酶A去饱和酶(SCD)、脂肪酸结合蛋白3(FABP3)、过氧化物酶体增殖物激活受体γ(PPARG)和分化抗原簇36(CD36)的相对表达量均无显著的一次线性或二次曲线关系(P>0.05)。综上,BHBA对BMECs活力的促进作用呈显著的二次曲线增加,即BHBA对BMECs活力呈显著浓度依赖关系;BHBA对细胞内乳脂肪的合成有提高的趋势。

关键词： β-羟丁酸; 乳腺上皮细胞; 乳脂肪合成相关基因; 奶牛

本文引用格式

常晨城 , 齐利枝 , 闫素梅 , 生冉 , 赵艳丽 . β-羟丁酸对奶牛乳腺上皮细胞内乳脂肪合成及其相关基因相对表达量的影响[J]. 动物营养学报, 2015 , 27(1) : 196 -203 . DOI: 10.3969/j.issn.1006-267x.2015.01.024

Abstract

This study was conducted to determine the effects of β-hydroxybutyric acid (BHBA) on cell viability, triacylglycerol (TAG) content, lipid droplet formation and relative expression levels of genes related to milk fat synthesis in bovine mammary epithelial cells (BMECs). The 3th passage cells were plated 1×105 cells/well in culture plates and DMEM/F12 medium containing 10% fetal bovine serum (FBS) was added to each well, then the plates were maintained in an incubator with 5% CO2 at 37 ℃. After 48 h, cells were randomly divided into 6 groups and cultured in DMEM/F12 medium containing different concentrations of BHBA, FBS in medium was replaced with 1 g/L fatty acid-free bovine serum albumin (BSA). The final concentrations of BHBA in reaction systems were 0 (control), 0.58, 1.16, 2.32, 4.64, 9.28 mmol/L, respectively. Continuous incubation in an incubator with 5% CO2 at 37 ℃ for 48 h. The results showed that BMECs proliferation [relative growth rate (RGR)] was significantly quadratic increased with BHBA concentration increasing (P=0.041), and BMECs proliferation of 0.58 to 4.64 mmol/L BHAB groups was high, but 9.28 mmol/L BHBA group was low. BHBA at low concentration (0.58 to 2.32 mmol/L) promoted lipid droplet formation, and the promoting effect of BHBA at high concentration (4.64 to 9.28 mmol/L) was weakened. No significant linear and quadratic relationships were found between BHBA concentration and TAG content (P>0.05). No significant linear and quadratic relationships were found between BHBA concentration and relative expression levels of genes related to milk fat synthesis (fatty acid synthetase, acetyl-CoA carboxylase α, stearyl coenzyme A desaturase, fatty acid binding protein 3, peroxisome proliferator-activated receptor γ and cluster of differentiation 36) (P>0.05). In conclusion, BMECs proliferation significantly quadratic increase with BHBA concentration increasing, namely BMECs proliferation has significant concentration-dependent relationship with BHBA concentration, and the milk fat synthesis has a improve trend by BHBA concentration.

Key words： β-hydroxybutyric acid; mammary epithelial cells; genes related to milk fat synthesis; dairy cows

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