&beta;-羟丁酸浓度对奶牛乳腺上皮细胞内乳蛋白合成相关基因表达量的影响

王新朋; 闫素梅; 齐利枝; 生冉; 赵艳丽

doi:10.3969/j.issn.1006-267x.2014.12.038

动物营养学报 >

2014 , Vol. 26 >Issue 12: 3836 - 3842

DOI: https://doi.org/10.3969/j.issn.1006-267x.2014.12.038

研究简报 Short Communications

β-羟丁酸浓度对奶牛乳腺上皮细胞内乳蛋白合成相关基因表达量的影响

王新朋 ,
闫素梅 ,
齐利枝 ,
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: 2014-06-27

网络出版日期: 2014-12-05

基金资助

奶业"973计划"(2011CB100800)

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Effects of β-Hydroxybutyric Acid on Expressions of Genes Involved in Milk Protein Synthesis in Bovine Mammary Epithelial Cells

WANG Xinpeng ,
YAN Sumei ,
QI Lizhi ,
SHENG Ran ,
ZHAO Yanli

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College of Animal Science, Inner Mongolia Agricultural University, Huhhot 010018, China

Received date: 2014-06-27

Online published: 2014-12-05

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摘要

本试验研究了不同浓度的β-羟丁酸(BHBA)对奶牛乳腺上皮细胞(BMEC)内乳蛋白合成相关基因表达量的影响.将传至第3代的BMEC以适宜的密度培养48 h后,随机分为6组,每组6个重复,每个重复1个培养孔.各组分别采用BHBA浓度为0(对照组)、0.58、1.16、2.32、4.64和9.28 mmol/L的培养液,置于37 ℃、5% CO₂培养箱中继续培养48 h.收集细胞,采用SYBR Green实时荧光定量PCR法测定乳蛋白合成相关基因的表达量.结果表明:随着BHBA浓度的增加,αs1-酪蛋白(CSN1S1)基因表达量呈显著的一元二次曲线增加(R²=0.931 6,P=0.018),其中以2.32~9.28 mmol/L组较高,尤以4.64 mmol/L BHBA组最高.随着BHBA浓度的增加,瘦素(leptin)基因表达量呈一定的一次线性降低趋势(R²=0.482 5,P=0.126),从数值上看,以0.58 mmol/L BHBA组最高.信号转导和转录激活因子5(STAT5)和哺乳动物雷帕霉素靶点(mTOR)基因表达量与BHBA浓度间的回归关系不显著(P=0.459,P=0.398).综合以上指标,BHBA的浓度为2.32~9.28 mmol/L可上调CSN1S1基因的表达,尤以BHBA浓度为4.64 mmol/L时对BMEC内乳蛋白合成的促进效果最好.

关键词： β-羟丁酸; 奶牛; 乳腺上皮细胞; 乳蛋白

本文引用格式

王新朋 , 闫素梅 , 齐利枝 , 生冉 , 赵艳丽 . β-羟丁酸浓度对奶牛乳腺上皮细胞内乳蛋白合成相关基因表达量的影响[J]. 动物营养学报, 2014 , 26(12) : 3836 -3842 . DOI: 10.3969/j.issn.1006-267x.2014.12.038

Abstract

The purpose of this study was to examine the effects of β-hydroxybutyric acid (BHBA) on expressions of genes involved in milk protein synthesis in bovine mammary epithelial cells (BMECs). The 3rd passage BMECs with proper densities were cultured for 48 h, then were divided into 6 groups with 6 replicates in each group and 1 culture pore per replicate. Culture mediums containing different concentrations of BHBA [0 (control group), 0.58, 1.16, 2.32, 4.64 and 9.28 mmol/L, respectively] were used in each group. Cells were continuous incubated in an incubator with 5% CO₂ at 37 ℃ for 48 h, then were collected for the analysis of expressions of milk protein and the expression of related genes in real-time fluorescent quantitative PCR with SYBR Green. The results showed as follows: with the increasing of BHBA concentration, the expression of CSN1S1 gene was significantly increased quadratically (R²=0.931 6, P=0.018), 2.32 to 9.28 mmol/L had better effect, and 4.64 mmol/L was optimal concentration. With the increasing of BHBA concentration, leptin gene expression was linearly down-regulated at a certain extent (R²=0.482 5, P=0.126), and 0.58 mmol/L was optimal concentration in value. With the increasing of BHBA concentration, signal transducer and activator of transcription 5 (STAT5) and mammalian target of rapamycin (mTOR) gene expression were not significantly changed (P=0.459, P=0.398). Taken together, the expression of CSN1S1 gene is up-regulated when cells areincubated with BHBA at 2.32 to 9.28 mmol/L, and the synthesis of milk proteins in BMECs has better effect when the concentration of BHBA is 4.64 mmol/L.

Key words： β-hydroxybutyric acid; dairy cow; bovine mammary epithelial cells; milk protein

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