基于GC-MS代谢组学技术对内脂素调控雏鸡采食机制的研究

doi:10.11843/j.issn.0366-6964.2017.07.009

摘要/Abstract

摘要：

旨在探索内脂素（Visfatin）调控家禽采食的分子机制，为完善家禽的食欲调控和能量平衡理论奠定基础。选取1日龄罗曼褐健康雏鸡20只，随机分为4个处理（C、L、M、H组）。常规饲粮饲养至10日龄进行下丘脑导管埋置，通过脑室注射技术对雏鸡下丘脑部位注射不同浓度的重组人内脂素（0、40、200、400 ng）。脑室注射Visfatin 60 min后，记录各处理组累积采食量，并采集下丘脑组织，通过GC-MS系统进行代谢组学检测，建立PLS-DA模型，并将差异代谢物输入KEGG数据库进行代谢通路的构建。结果表明，脑室注射Visfatin 60 min后，M组和H组的雏鸡累积采食量与C组相比均显著升高（P<0.05）。在对雏鸡下丘脑进行GC-MS代谢组学分析后，处理组与对照组间共鉴定获得73个差异性代谢物，富集到51条通路上。脑室注射Visfatin后下丘脑产生的差异性代谢物与氨基酸、糖和脂代谢有着密切的联系，其中脂肪酸合成通路被显著富集。结果提示，中枢营养物质下降导致稳态发生改变可能是Visfatin发挥促食作用的主要机制。

Abstract:

The objectives of the study were to explore the molecular mechanism of Visfatin regulating feed intake, and to further improve the theory of appetite regulation and energy balance in poultry.In this study, 20 healthy Roman Brown chicks at the age of 1 day old were randomly divided into 4 groups(group C, L, M, H). When the chicks grew up to the age of 10 days old fed with conventional commercial diet, they were intracerebroventricularly injected with various concentrations of human recombinant Visfatin(0, 40, 200, 400 ng) in hypothalamus, respectively. The cumulative feed intake was recorded after injection for 60 min. The chicks were then euthanized and the hypothalamus were collected. Metabolomics analysis was performed through GC-MS system. The PLS-DA model was built and the metabolic pathways of differential metabolites were constructed by KEGG. The results showed that the cumulative feed intakes of chicks in M and H groups were increased significantly compared to that in C group (P <0.05). A total of 73 differential metabolites, which were enriched in 51 pathways, were identified by GC-MS analysis.The differential metabolites in hypothalamus were closely related to carbohydrate, lipid and amino acid metabolism, and fatty acid biosynthesis pathway was one of the most significantly enriched pathways. The main reason for Visfatin promoting food intake is the change of steady state due to decrease of central nutrients.

中图分类号:

S831.2

刘雪莲, 张盼盼, 田方圆, 李准, 李转见, 韩瑞丽, 孙桂荣, 蒋瑞瑞, 刘小军, 康相涛, 田亚东. 基于GC-MS代谢组学技术对内脂素调控雏鸡采食机制的研究[J]. 畜牧兽医学报, 2017, 48(7): 1251-1259.

LIU Xue-lian, ZHANG Pan-pan, TIAN Fang-yuan, LI Zhun, LI Zhuan-jian, HAN Rui-li, SUN Gui-rong, JIANG Rui-rui, LIU Xiao-jun, KANG Xiang-tao, TIAN Ya-dong. Research of Visfatin Regulating Chicks Feed Intake Based on GC-MS Metabolomics[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(7): 1251-1259.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2017.07.009

https://www.xmsyxb.com/CN/Y2017/V48/I7/1251

参考文献

[1] 张盼盼,商鹏飞,田方圆,等. 鸡Visfatin蛋白的原核表达、纯化及其活性研究[J]. 畜牧兽医学报,2016, 47(9):1785-1794.
ZHANG P P, SHANG P F, TIAN F Y, et al. Prokaryotic expression,purification and bioactivity identification of recombinant chicken visfatin protein[J]. Acta Veterinaria et Zootechnica Sinica,2016, 47(9):1785-1794. (in Chinese)
[2] 于继英,黄美珍. 畜禽采食量的调节[J]. 中国畜牧兽医, 2005, 32(9):6-8.
YU J Y, HUANG M Z. Effect of dietary oliseeds supplementation on milk regulation of feed intake[J]. China Animal Husbandry and Veterinary Medicine, 2005, 32(9):6-8. (in Chinese)
[3] FUKUHARA A, MATSUDA M, NISHIZAWA M, et al. Visfatin:a proteinsecreted by visceral fat that mimics the effects of insulin[J]. Science, 2005, 307(5708):426-430.
[4] REVOLLO J R, KÖMER A, MILLS K F, et al. Nampt/PBEF/visfatin regulates insulin secretion in β cells as a systemic NAD biosynthetic enzyme[J]. Cell Metab, 2007, 6(5):363-375.
[5] DIOT M, REVERCHON M, RAMÉ C, et al. Expression and effect of NAMPT (visfatin) on progesterone secretion in hen granulosa cells[J]. Reproduction, 2015, 150(1):53-63.
[6] REVERCHON M, CORNUAU M, CLOIX L, et al. Visfatin is expressed in human granulosa cells:regulation by metformin through AMPK/SIRT1 pathways and its role in steroidogenesis[J]. Mol Hum Reprod, 2013, 19(5):313-326.
[7] REVERCHONM,RAMÉ C, BERTOLDO M, et al. Adipokines and the female reproductive tract[J]. Int J Endocrinol, 2014,2014:232454.
[8] 温宇,杨姗姗,刘婧,等. 性激素对3T3-L1脂肪细胞Visfatin表达的影响[J]. 中国动脉硬化杂志, 2012, 20(10):871-875.
WEN Y, YANG S S, LIU J, et al. Regulation of sex steroid hormone on Visfatin expression in adipocytes and preadipocytes[J]. Chinese Journal of Arteriosclerosis, 2012, 20(10):871-875.(in Chinese)
[9] MOSCHEN A R, GERNER R R, TILG H. Pre-B cell colony enhancing factor/NAMPT/visfatin in inflammation and obesity-related disorders[J]. Curr Pharm Des, 2010, 16(17):1913-1920.
[10] ZHANG L Q, HERUTH D P, YE S Q. Nicotinamide phosphoribosyltransferase in human diseases[J]. J Bioanal Biomed, 2011, 3(1):13-25.
[11] CHEN M P, CHUNG F M, CHANG D M, et al. Elevated plasma level of visfatin/pre-B cell colony-enhancing factor in patients with type 2 diabetes mellitus[J]. J Clin Endocrinol Metab, 2006, 91(1):295-299.
[12] OGNJANOVIC S, BRYANT-GREENWOOD G D. Pre-B-cell colony-enhancing factor, a novel cytokine of human fetal membranes[J]. Am J Obstet Gynecol, 2002,187(4):1051-1058.
[13] YE S Q, SIMON B A, MALONEY J P, et al. Pre-B-cell colony-enhancing factor as a potential novel biomarker in acute lung injury[J]. Am J Respir Crit Care Med, 2005, 171(4):361-370.
[14] CLINE M A, NANDAR W, PRALL B C, et al. Central visfatin causes orexigenic effects in chicks[J]. Behav Brain Res, 2008, 186(2):293-297.
[15] BRUNETTI L, RECINELLA L, NISIO C, et al. Effects of visfatin/PBEF/NAMPT on feeding behaviour and hypothalamic neuromodulators in the rat[J]. J Biol Regul Homeost Agents, 2012, 26(2):295-302.
[16] NICHOLSON J K, LINDON J C. Systems biology:metabonomics[J]. Nature, 2008, 455(7216):1054-1056.
[17] FIEHN O, KOPKA J, DÖRMANN P, et al. Metabolite profiling for plant functional genomics[J]. Nat Biotechnol, 2000, 18(11):1157-1161.
[18] GAO X F, PUJOS-GUILLOT E,SÉBÉDIO J L. Development of a quantitative metabolomic approach to study clinical human fecal water metabolome based on trimethylsilylation derivatization and GC/MS analysis[J]. Anal Chem, 2010, 82(15):6447-6456.
[19] WOLTHERS B G, KRAAN G P B. Clinical applications of gas chromatography and gas chromatography-mass spectrometry of steroids[J]. J Chromatogr A, 1999, 843(1-2):247-274.
[20] KABURAGI Y, FUKABORI Y, TAKAHASHI E, et al. The metabolism of testosterone in the central nervous system (1). Analysis of testosterone metabolites in the anterior pituitary and hypothalamus using gas chromatography-mass spectrometry (GC-MS), and subcellular localization of testosterone converting enzyme[J]. Nihon Naibunpi Gakkai Zasshi, 1984, 60(11):1314-1327.
[21] LIN J C, SU M M, WANG X Y, et al. Original paper multiparametric analysis of amino acids and organic acids in rat brain tissues using GC/MS[J]. J Sep Sci, 2008, 31(15):2831-2838.
[22] LÓPEZ M, LAGE R, SAHA A K, et al. Hypothalamic fatty acid metabolism mediates the orexigenic action of ghrelin[J]. Cell Metab, 2008, 7(5):389-399.
[23] OOMURA Y, NAKAMURA T, SUGIMORI M, et al. Effect of free fatty acid on the rat lateral hypothalamic neurons[J]. Physiol Behav, 1975, 14(4):483-486.
[24] OBICI S, FENG Z H, MORGAN K, et al. Central administration of oleic acid inhibits glucose production and food intake[J]. Diabetes, 2002, 51(2):271-275.
[25] OBICI S, FENG Z H,ARDUINI A, et al. Inhibition of hypothalamic carnitine palmitoyltransferase-1 decreases food intake and glucose production[J]. Nat Med, 2003, 9(6):756-761.
[26] CRUCIANI-GUGLIELMACCI C, HERVALET A, DOUARED L, et al. Beta oxidation in the brain is required for the effects of non-esterified fatty acids on glucose-induced insulin secretion in rats[J]. Diabetologia, 2004, 47(11):2032-2038.
[27] FUJIWARA K, MAEKAWA F, DEZAKI K, et al. Oleic acid glucose-independently stimulates glucagon secretion by increasing cytoplasmic Ca²⁺via endoplasmic reticulum Ca²⁺ release and Ca²⁺influx in the rat islet α-cells[J]. Endocrinology, 2007, 148(5):2496-2504.
[28] ASECHI M, KURAUCHI I, TOMONAGA S, et al. Relationships between the sedative and hypnotic effects of intracerebroventricular administration of L-serine and its metabolites, pyruvate and the derivative amino acids contents in the neonatal chicks under acute stressful conditions[J]. Amino Acids, 2008, 34(1):55-60.
[29] GIBALA M J, YOUNG M E, TAEGTMEYER H. Anaplerosis of the citric acid cycle:role in energy metabolism of heart and skeletal muscle[J]. Acta physiol Scand, 2000, 168(4):657-665.
[30] HE W H, MIAO F J P, LIN D C H, et al. Citric acid cycle intermediates as ligands for orphan G-protein-coupled receptors[J]. Nature, 2004, 429(6988):188-193.