杜仲叶对绵羊肝脏糖代谢及其相关基因表达的影响

高燕, 杨改青, 王林枫, 杜庆鑫, 王文辉, 付彤, 廉红霞, 高腾云, 李明

doi:10.3969/j.issn.1006-267x.2019.06.046

动物营养学报 >

2019 , Vol. 31 >Issue 6: 2854 - 2864

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.06.046

饲料科学与技术 Feed science and technology

杜仲叶对绵羊肝脏糖代谢及其相关基因表达的影响

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1. 河南农业大学牧医工程学院, 郑州 450046;
2. 河南农业大学现代实验技术与管理中心, 郑州 450002;
3. 中国林业科学研究院经济林研究开发中心, 郑州 450003;
4. 河南省种牛遗传性能测定中心, 郑州 450002

高燕(1995-),女,河南新安人,硕士研究生,从事营养生理与调控的研究。E-mail:gaoyli1995@163.com

收稿日期: 2018-11-28

网络出版日期: 2019-06-17

基金资助

国家奶牛产业技术体系专项（CARS-36）；河南省现代农业产业技术体系专项基金（S2013-08-01）

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Effects of Eucommia ulmoides Leaves on Glucose Metabolism and Related Gene Expression in Sheep Liver

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1. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China;
2. Advanced Experimental Techniques and Management Centre, Henan Agricultural University, Zhengzhou 450002, China;
3. Economic Forestry Research and Development Center, Chinese Academy of Forestry Sciences Zhengzhou 450003, China;
4. Henan Genetic Performance Measurement Center for Breeding Cattle, Zhengzhou 450002, China

Received date: 2018-11-28

Online published: 2019-06-17

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摘要

本试验旨在研究杜仲叶（EUL）对绵羊肝脏糖代谢及其相关基因表达的影响。随机选取6~8月龄、体重35~40 kg的绵羊（杜泊羊♂×湖羊♀）12只，平均分为2组，分别为对照组（CTL组）和EUL组（饲粮中添加10% EUL），每组6只。预试期10 d，正试期30 d。采集血液，测定糖代谢相关指标；采集肝脏组织，用于转录组学测序、糖代谢相关酶及核转录因子的mRNA及蛋白表达量分析。结果表明：1）与CTL组相比，EUL组血浆葡萄糖（GLU）、血清胰高血糖素（GC）含量显著降低（P<0.05），血清胰岛素（INS）含量显著升高（P<0.05）。2）转录组学检测结果表明，差异显著基因与糖代谢和能量代谢密切相关，且其在与糖代谢相关的通路中显著富集。3）荧光定量PCR检测结果表明，与CTL组相比，EUL组肝脏胰岛素受体（INSR）及胰岛素受体底物2（IRS2）的mRNA表达量极显著升高（P<0.01），胰高血糖素受体（GCGR）的mRNA表达量极显著降低（P<0.01）；糖酵解关键酶磷酸果糖激酶（PFKL）的mRNA表达量显著上调（P<0.05）；糖异生相关酶葡萄糖-6-磷酸酶（G6Pase）mRNA表达量极显著下调（P<0.01），磷酸烯醇式丙酮酸羧激酶1（PEPCK1）的mRNA表达量显著下调（P<0.05）；调控糖异生的叉头转录因子1（FoxO1）和环磷酸腺苷应答元件结合蛋白（CREB）的mRNA表达量均极显著下调（P<0.01）；肝糖原合成关键酶糖原合酶2（Gys2）的mRNA表达量极显著上调（P<0.01）。4）Western blot结果显示，与CTL组相比，EUL组INSR蛋白表达量无显著变化（P>0.05）；FoxO1、G6Pase和PEPCK1蛋白表达量均极显著下调（P<0.01）。综上所述，EUL显著影响绵羊机体的糖代谢，提高糖酵解关键酶基因的表达，抑制糖异生相关转录因子及酶基因的表达，并促进肝糖原的合成，进而降低血糖。

关键词： 杜仲叶; 糖代谢; 肝脏; 基因表达; 绵羊

本文引用格式

高燕, 杨改青, 王林枫, 杜庆鑫, 王文辉, 付彤, 廉红霞, 高腾云, 李明 . 杜仲叶对绵羊肝脏糖代谢及其相关基因表达的影响[J]. 动物营养学报, 2019 , 31(6) : 2854 -2864 . DOI: 10.3969/j.issn.1006-267x.2019.06.046

Abstract

In order to research the effects of Eucommia ulmoides leaves (EUL) on glucose metabolism and related genes expression in sheep live, 12 healthy sheep (dorper sheep♂×Hu lambs♀) with the similar age (6 to 8-month-old) and body weight (between 35 to 40 kg) were selected and divided into two groups randomly and equally (6 sheep in each group), which named control group (CTL group) and EUL group (diet contained 10% of EUL). The adaption period was 7 days, and the experiment period was 30 days. At the end of experiment, blood samples were collected to determine related biochemical indexes and hormones. After the experiment, all sheep were slaughtered and liver tissues were collected to transcriptome sequence, determine genes and protein expression levels for enzymes and factors involved in glucose metabolism. The results showed as follows:1) compared with CTL group, plasma glucose and serum glucagon contents in EUL group significantly decreased (P<0.05), whereas serum insulin content increased significantly (P<0.05). 2) The results of transcriptome analysis showed that the differentially significant genes related to glucose metabolism and energy metabolism, and significantly enriched in the glucose metabolic pathways. 3) Fluorescent quantitative PCR results showed that the mRNA expression levels of insulin receptor (INSR) and insulin receptor substrate 2 (IRS2) increased significantly (P<0.01), the mRNA expression level of glucagon receptor (GCGR) dropped markedly (P<0.01); the phosphofructokinase (PFKL), the key enzyme in the glycolytic pathway, mRNA expression level up-regulated notably (P<0.05); on the gluconeogenesis pathway, the mRNA expression level of glucose-6-phosphatase (G6Pase) decreased significantly (P<0.01) and the mRNA expression level of phosphoenolpyruvate carboxykinase 1 (PEPCK1) decreased significantly (P<0.05); the mRNA expression levels of fork head transcription factor 1 (FoxO1) and cyclic adenosine monophosphate response element binding protein (CREB) decreased significantly (P<0.01); furthermore, glycogen synthase 2 (Gys2), which promoted glycogen synthesis, mRNA expression increased significantly (P<0.01). 4) Western blot results showed that compared with CTL group, the protein expression level of INSR in EUL group had no significant change (P>0.05), but the protein expression levels of FoxO1、G6Pase and PEPCK1 decreased significantly (P<0.01). In conclusion, EUL greatly affects glucose metabolism of sheep, increases the expression of the key enzyme genes for glycolysis, down-regulates the expression of gluconeogenesis related nuclear transcription factors and enzymes, promotes the synthesis of liver glycogen, and then decreases blood glucose.

Key words： Eucommia ulmoides leaves; glucose metabolism; liver; gene expression; sheep

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