宫内生长受限湖羊新生羔羊的血浆代谢组学研究

doi:10.13304/j.nykjdb.2021.1060

摘要/Abstract

摘要：

湖羊是我国珍贵的肉毛兼用的绵羊品种，并具有繁殖率高和宜舍饲等特点。在生产中，往往因其多胎性易导致妊娠期湖羊营养不良，造成新生羔羊宫内生长受限（intrauterine growth retardation，IUGR）。应用代谢组学一维氢谱核磁共振技术（¹H-nuclear magnetic resonance，¹H-NMR）研究新生羔羊的血浆代谢谱。根据出生体质量筛选出14只湖羊新生羔羊，分为正常出生体质量组（normal birth weight，NBW）和IUGR组（n=7），采用¹H-NMR分析其血液样品。将代谢谱数据进行统计分析，并对鉴定的代谢物进行表达差异分析以及代谢途径归属。NBW组和IUGR组新生羔羊血浆样品共获得32种代谢物，16种代谢物在2组间差异表达，IUGR组有10个代谢物含量显著低于NBW组（P<0.05），6个代谢物含量显著高于NBW组（P<0.05）。2组间差异显著的代谢通路主要为氨基酸、碳水化合物、脂类物质等代谢通路。应用基于¹H-NMR的代谢组学及生物信息学分析方法，确定了IUGR新生羔羊与正常羔羊血液代谢物之间存在差异，为进一步深入研究IUGR的分子机制奠定了基础。

关键词: 湖羊, 代谢组学, 一维氢谱核磁共振技术, 胎儿宫内生长受限, 代谢通路

Abstract:

As a precious breed in China， Hu-sheep is a good duel-purpose breed both for mutton and wool， which have precociousness and hyper-prolificacy. Because of multiple pregnancy， it is easy to lead to malnutrition during pregnancy， resulting in intrauterine growth restriction （IUGR） of newborn lambs. In order to analyze the plasma metabolic profiles of neonatal lambs with IUGR and compare them with controls to define the metabolic patterns associated with IUGR， 14 neonatal lambs of Hu-sheep were screened according to birth weight， and then allotted to two groups， normal birth weight （NBW） and IUGR groups （n=7）. ¹H-nuclear magnetic resonance （¹H-NMR） based metabolomics approach of the plasma samples collected from the two groups was obtained. The results was used to screen differential metabolites and analyze related metabolic pathways. The plasma samples of the NBW and IUGR groups were detected by ¹H-NMR， and 32 metabolites were obtained. 16 metabolites were differentially expressed between the 2 groups. Compared with NBW group， the concentrations of 10 metabolites were significantly lower in IUGR group （P<0.05）， and the concentrations of 6 metabolites were significantly upper （P<0.05）. The significant metabolic pathways of those metabolites were amino acids， carbohydrates， and lipids pathways. This study indicated that ¹H-NMR based on metabolomics and bioinformatics analysis could be used to characterize changes in metabolomics between neonates lambs of IUGR and controls， which provided foundation for further in-depth molecular mechanism studies on IUGR.

Key words: Hu-sheep, metabolomics, ¹H-NMR, IUGR, metabolic pathway

中图分类号:

S826

孙玲伟, 何孟纤, 戴建军, 吴彩凤, 张德福, 林月霞. 宫内生长受限湖羊新生羔羊的血浆代谢组学研究[J]. 中国农业科技导报, 2022, 24(7): 123-131.

Lingwei SUN, Mengxian HE, Jianjun DAI, Caifeng WU, Defu ZHANG, Yuexia LIN. Metabolomics in Neonatal Lambs of Hu-sheep with Intrauterine Growth Retardation[J]. Journal of Agricultural Science and Technology, 2022, 24(7): 123-131.

图/表 4

图1 NBW组和IURG组血浆样品代表性的600 MHz 1H-NMR图谱注：1—低密度/极低密度脂蛋白；2—异戊酸；3—亮氨酸；4—缬氨酸；5—3-羟丁酸；6—乳酸； 7—丙氨酸；8—乙酸盐；9—2-羟基异戊酸酯；10—N-乙酰甘露糖胺；11—N-乙酰半胱氨酸；12—谷氨酰胺；13—丙酮；14—脲基丙酸；15—柠檬酸；16—肌肽；17.异柠檬酸；18—肌酸；19—丙二酸；20—甘油磷酸胆碱；21—胆碱；22—磷酸胆碱；23—β-葡萄糖；24—甜菜碱；25—α-葡萄糖；26—甘氨酸；27—酪氨酸；28—1-甲基组氨酸；29—3-甲基组氨酸；30—苯丙氨酸；31—尿甘酸；32—甲酸。

Fig. 1 Representative 600 MHz 1H NMR spectra of plasma samples from NBW and IURG groupsNote：1—Low/very low density lipoprotein；2—Isovalerate；3—L-leucine；4—L-valine；5—3-hydroxybutyrate； 6—Lactate； 7—Alanine； 8—Acetate； 9—2-hydroxyisovalerate；10—N-acetylmannosamine；11—N-acetylcysteine；12—Glutamine；13—Acetone； 14—Ureidopropionic acid； 15—Citrate； 16—Carnosine；17—Isocitrate； 18—Creatine； 19—Malonate； 20—Glycerophosphocholine； 21—Choline； 22—O-phosphocholine； 23—?β?-glucose； 24—Betaine； 25—?α?-glucose； 26—Glycine； 27—Tyrosine； 28—1-methylhistidine； 29—3-methylhistidine； 30—Phenylalanine； 31—Urocanate； 32—Formate.

图2 NBW组和IUGR组新生羔羊血浆样品的PCA得分

Fig. 2 PCA score of plasma form newborn lambs in the NBW group and the IUGR group

图3 2组新生羔羊血浆样品的PLS-DA分析

Fig. 3 PLS-DA plot of newborn lambs in the NBW and the IUGR groups

表1 1H-NMR检测的两组间有显著变化的代谢物峰强度

Table 1 Peak intensity of significantly changed metabolites between the two groups based on 1H-NMR

编号 No	代谢物 Metabolites	NBW组	IUGR组	分类 Classification
1	低密度脂蛋白/极低密度脂蛋白 LDL/VLDL	71.81±14.71	99.19±24.08^*	脂类 Lipid
2	异戊酸 Isovalerate	19.38±4.01	13.84±4.34^*	脂类 Lipid
3	亮氨酸 Leucine	14.08±1.80	10.68±2.81^*	氨基酸 Amino acids
4	缬氨酸 Valine	13.80±2.55	9.44±2.13^**	氨基酸 Amino acids
5	乳酸 Lactate	77.45±11.94	50.46±7.22^*	糖类 Carbohydrate
6	丙氨酸 Alanine	16.21±1.42	10.36±2.51^**	氨基酸 Amino acids
7	2-羟基异戊酸 2-hydroxyisovalerate	10.43±2.20	14.19±1.07^*	脂类 Lipid
8	N-乙酰半胱氨酸 N-acetylcysteine	23.14±1.92	14.60 ± 2.65^*	氨基酸 Amino acids
9	甘油磷酸胆碱Glycerophosphocholine	22.37±1.95	32.02±4.53^*	脂类 Lipid
10	胆碱 Choline	87.20±4.03	145.20±22.73^*	脂类 Lipid
11	磷酸胆碱 Phosphocholine	111.61±13.03	185.33±49.41^**	脂类 Lipid
12	甜菜碱 Betaine	102.09±6.57	192.78±6.26^**	糖类 Carbohydrate
13	β-葡萄糖 β-glucose	87.56±10.36	61.47±4.95^*	糖类 Carbohydrate
14	α-葡萄糖 α-glucose	50.75±4.94	31.79±3.61^*	糖类 Carbohydrate
15	酪氨酸 Tyrosine	2.28±0.22	1.02±0.20^*	氨基酸 Amino acids
16	3-甲基组氨酸 3-methylhistidine	1.74±0.27	0.86±0.19^*	氨基酸 Amino acids

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