基于液相色谱-质谱联用技术分析鸡胚发育期间血清代谢物的变化  

Liquid chromatography-mass spectrometry-based analysis of change of serum metabolite during development of chick embryo

作　　者：彭梦玲 胡文业 王菊花[1] 张粟子 周杰[1] PENG Meng-ling;HU Wen-ye;WANG Ju-hua;ZHANG Su-zi;ZHOU Jie(College of Animal Science and Technology,Anhui Agricultural University,Hefei 230036,Anhui Province,China)

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036

出　　处：《中国生物制品学杂志》2020年第8期934-939,共6页Chinese Journal of Biologicals

基　　金：安徽省自然科学基金(1908085QC144);安徽农业大学青年基金重点项目(2018zd22)。

摘　　要：目的应用液相色谱-质谱联用技术(liquid chromatograph-mass spectrometer,LC-MS)分析肉鸡胚胎发育期间血清代谢物的变化。方法将120枚罗氏308肉鸡种蛋随机分为2组,分别于孵化的第14天(E14)和出壳1日龄(H1)收集血清样品。应用LC-MS和高分辨串联质谱SYNAPT G2-XS QTOF,结合主成分分析(principal component analysis,PCA)、偏最小二乘判别分析(partial least squares-discriminate analysis,PLS-DA)、变量投影重要性指标(variable important for the projection,VIP)确定差异代谢产物,并采用代谢物富集分析(metabolite set enrichment analysis,MESA)法分析差异代谢产物涉及的主要代谢途径。结果与E14比较,38个血清代谢物于H1发生显著变化,其中1个显著升高,37个显著下降。差异代谢物主要富集于22个代谢途径中,其中显著富集于蛋白质合成途径的差异代谢产物主要有L-苏氨酸、L-亮氨酸、L-异亮氨酸和L-甲硫氨酸;显著富集于糖异生途径的差异代谢产物主要有L-乳酸、α-酮戊二酸和β-D-葡萄糖;显著富集于缬氨酸、亮氨酸和异亮氨酸降解途径的差异代谢物主要有L-异亮氨酸、L-亮氨酸和脂酰胺。与E14比较,富集于以上代谢途径中的主要差异代谢产物于H1的含量均显著降低。结论鸡胚E14和H1血清代谢物存在显著差异,这些差异代谢物对鸡胚发育有一定影响,可作为日粮添加剂从雏鸡1日龄开始添加,从而满足肉鸡生长发育需要、改善肉鸡生产性能、节约蛋白饲料并降低成本。。Objective To analyze the changes of serum metabolites during development of chick embryo by liquid chromatography-mass spectrometry(LC-MS).Methods A total of 120 fertilized eggs from Ross 308 hens were randomly divided into two groups,20 for each in triplicates.Serum samples were collected on day 14 after incubation(E14)and day 1 after being hatched(H1).LC-MS quadrapole time of flight system(LC/MS-QTOF)combined with principal component analysis(PCA),partial least squares-discriminate analysis(PLS-DA)and variable important for the projection(VIP)were applied to determine the differentially changed metabolites,and metabolite set enrichment analysis(MSEA)to analyze the pathways involved in.Results Thirty-eight serum metabolites showed significant changes on H1 as compared with those on E14,of which one increased significantly and 37 decreased significantly.The differential metabolites were mainly enriched in 22 metabolism pathways,of which L-threonine,L-leucine,L-isoleucine and Lmethionine were mainly enriched in protein synthesis pathway,while L-lactic acid,α-ketoglutarate andβ-D-glucose in gluconeogenesis pathway,and L-isoleucine,L-leucine and lipoamide in degradation pathway of valine,leucine and isoleucine.However,all the contents of the differential metabolites enriched in various metabolic pathways on H1 decreased significantly as compared with those on E14.Conclusion The metabolites in sera on E14 and H1 showed significant difference,which showed a certain effect on the development of chick embryo.The metabolites may be used as the additives of daily diet of chicks starting from H1 so as to meet the requirement for growth of broilers,improve the production performance of broilers,save the protein forage and decrease the cost.

关 键 词：鸡胚 血清 代谢组学 液相色谱-质谱联用技术 

分 类 号：R373.23[医药卫生—病原生物学] Q75[医药卫生—基础医学]