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长期给泌乳山羊饲喂高谷物日粮对乳腺基因表达和DNA甲基化调控乳脂肪生成及成分的负面影响。

Negative effects of long-term feeding of high-grain diets to lactating goats on milk fat production and composition by regulating gene expression and DNA methylation in the mammary gland.

作者信息

Tian Ping, Luo Yanwen, Li Xian, Tian Jing, Tao Shiyu, Hua Canfeng, Geng Yali, Ni Yingdong, Zhao Ruqian

机构信息

Key Laboratory of Animal Physiology & Biochemistry, Nanjing Agricultural University, Nanjing, 210095 People's Republic of China.

College of Veterinary Medicine, Northwest A and F University, Yangling, Shannxi China.

出版信息

J Anim Sci Biotechnol. 2017 Oct 1;8:74. doi: 10.1186/s40104-017-0204-2. eCollection 2017.

Abstract

BACKGROUND

It is well known that feeding a high concentrate (HC) diet to lactating ruminants likely induces subacute ruminal acidosis (SARA) and leads to a decrease in milk fat production. However, the effects of feeding a HC diet for long periods on milk fatty acids composition and the mechanism behind the decline of milk fat still remains poorly understood. The aim of this study was to investigate the impact of feeding a HC diet to lactating dairy goats on milk fat yield and fatty acids composition with an emphasis on the mechanisms underlying the milk fat depression. Seventeen mid-lactating dairy goats were randomly allocated to three groups. The control treatment was fed a low-concentrate diet (35% concentrate,  = 5, LC) and there were two high-concentrate treatments (65% concentrate, HC), one fed a high concentrate diet for a long period (19 wks,  = 7, HL); one fed a high concentrate diet for a short period of time (4 wk,  = 5, HS). Milk fat production and fatty acids profiles were measured. In order to investigate the mechanisms underlying the changes in milk fat production and composition, the gene expression involved in lipid metabolism and DNA methylation in the mammary gland were also analyzed.

RESULTS

Milk production was increased by feeding the HC diet in the HS and HL groups compared with the LC diet ( < 0.01), while the percentage of milk fat was lower in the HL ( < 0.05) but not in the HS group. The total amount of saturated fatty acids (SFA) in the milk was not changed by feeding the HC diet, whereas the levels of unsaturated fatty acids (UFA) and monounsaturated fatty acids (MUFA) were markedly decreased in the HL group compared with the LC group ( < 0.05). Among these fatty acids, the concentrations of C15:0 ( < 0.01), C17:0 ( < 0.01), C17:1 ( < 0.01), C18:1n-9c ( < 0.05), C18:3n-3r ( < 0.01) and C20:0 ( < 0.01) were markedly lower in the HL group, and the concentrations of C20:0 ( < 0.05) and C18:3n-3r ( < 0.01) were lower in the HS group compared with the LC group. However, the concentrations of C18:2n-6c ( < 0.05) and C20:4n-6 ( < 0.05) in the milk fat were higher in the HS group. Real-time PCR results showed that the mRNA expression of the genes involved in milk fat production in the mammary gland was generally decreased in the HL and HS groups compared with the LC group. Among these genes, , & , , , , and were down-regulated in the mammary gland of the HL group ( < 0.05), and the expressions of , and mRNA were markedly decreased in the HS goats compared with the LC group ( < 0.05). In contrast to the gene expression, the level of DNA methylation in the promoter regions of the and genes was increased in the HL group compared with the LC group ( < 0.05). The levels of ACSL1 protein expression and FAS enzyme activity were also decreased in the mammary gland of the HL compared with the LC group ( < 0.05).

CONCLUSIONS

Long-term feeding of a HC diet to lactating goats induced milk fat depression and FAs profile shift with lower MUFAs but higher SFAs. A general down-regulation of the gene expression involved in the milk fat production and a higher DNA methylation in the mammary gland may contribute to the decrease in milk fat production in goats fed a HC diet for long time periods.

摘要

背景

众所周知,给泌乳反刍动物饲喂高浓度(HC)日粮可能会引发亚急性瘤胃酸中毒(SARA),并导致乳脂肪产量下降。然而,长期饲喂HC日粮对乳脂肪酸组成的影响以及乳脂肪下降背后的机制仍知之甚少。本研究的目的是调查给泌乳奶山羊饲喂HC日粮对乳脂肪产量和脂肪酸组成的影响,重点是乳脂肪降低的潜在机制。17只处于泌乳中期的奶山羊被随机分为三组。对照处理饲喂低浓度日粮(35%精料,n = 5,LC),有两个高浓度处理组(65%精料,HC),一组长期饲喂高浓度日粮(19周,n = 7,HL);一组短期饲喂高浓度日粮(4周,n = 5,HS)。测定了乳脂肪产量和脂肪酸谱。为了研究乳脂肪产量和组成变化的潜在机制,还分析了乳腺中参与脂质代谢的基因表达和DNA甲基化情况。

结果

与LC日粮相比,HS组和HL组饲喂HC日粮可提高产奶量(P < 0.01),而HL组乳脂肪百分比降低(P < 0.05),但HS组未降低。饲喂HC日粮不会改变乳中饱和脂肪酸(SFA)的总量,而HL组与LC组相比,不饱和脂肪酸(UFA)和单不饱和脂肪酸(MUFA)水平显著降低(P < 0.05)。在这些脂肪酸中,HL组中C15:0(P < 0.01)、C17:0(P < 0.01)、C17:1(P < 0.01)、C18:ln-9c(P < 0.05)、C18:3n-3r(P < 0.01)和C20:0(P < 0.01)的浓度显著降低,与LC组相比,HS组中C20:0(P < 0.05)和C18:3n-3r(P < 0.01)的浓度较低。然而,HS组乳脂肪中C18:2n-6c(P < 0.05)和C20:4n-6(P < 0.05)的浓度较高。实时PCR结果显示,与LC组相比,HL组和HS组乳腺中参与乳脂肪合成的基因mRNA表达普遍降低。在这些基因中,HL组乳腺中ACACA、FASN、SCD、FABP3、DGAT1、FATP2和LPIN1下调(P < 0.05),与LC组相比,HS组山羊中ACACA、FASN和SCD mRNA表达显著降低(P < 0.05)。与基因表达相反,与LC组相比,HL组中ACACA和FASN基因启动子区域的DNA甲基化水平升高(P < 0.05)。与LC组相比,HL组乳腺中ACSL1蛋白表达水平和FAS酶活性也降低(P < 0.05)。

结论

长期给泌乳山羊饲喂HC日粮会导致乳脂肪降低和脂肪酸谱改变,MUFA含量降低但SFA含量升高。乳腺中参与乳脂肪合成的基因表达普遍下调以及DNA甲基化水平升高可能是长期饲喂HC日粮的山羊乳脂肪产量降低的原因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/68ec/5623059/b39d26c28ba8/40104_2017_204_Fig1_HTML.jpg

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