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

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.%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 metaboliteswere 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.