基于GC-MS策略的奶牛热应激血液代谢组学研究

doi:10.11843/j.issn.0366-6964.2015.08.011

摘要/Abstract

摘要：

为了系统揭示热应激对奶牛血液代谢物及其代谢通路的影响。本研究根据奶牛热应激的判定标准，选择10头处于热应激状态的荷斯坦高产泌乳牛为热应激组，至次日凌晨直肠温度和呼吸频率恢复到正常范围时，再次被视为恢复组。各组奶牛尾静脉采血，制备血浆。然后采用GC-MS代谢组学技术，结合模式识别策略寻找两组奶牛血液差异代谢物，并将差异代谢物输入KEGG数据库进行代谢通路的构建与功能分析。结果表明，8个内源性代谢物可作为奶牛热应激的潜在生物标志物。其中，葡萄糖、α-亚麻酸、亚油酸、甘油、棕榈酸、β-羟丁酸和甘氨胆酸盐含量在热应激过程中显著降低，而乳酸含量显著升高。提示热应激加剧了奶牛能量负平衡状态，主要通过增强脂肪酸氧化和甘油分解代谢途径活性，抑制糖酵解过程进行调节和应答，在此过程中伴有肝功能障碍的生理现象。研究结果可为进一步阐明奶牛热应激的生理机制提供科学依据。

关键词: 热应激, 奶牛, 代谢组学, GC-MS, 血液代谢物

Abstract:

The study aimed to reveal the effects of heat stress on dairy cow blood metabolites and its metabolic pathways in a systematic way.According to the criteria for evaluating cow heat stress，10 high milk yield Holstein cows in heat stress status were selected as heat stress group，these cows were regarded as recovery group again until their rectal temperature and respiratory rate returned to the normal range at next morning.Blood samples of each group were collected via tail vein and the plasm were separated.And then we developed and applied a gas chromatography-mass spectrometry GC-MS metabolomics protocol combined with pattern recognition approaches to search differential metabolites between the heat stress and recovery groups，and the differential metabolites were analyzed by KEGG for reconstructing the metabolic pathway and functional analysis.8 metabolites were selected as heat stress potential biomakers and indentified.These biomarkers included alpha-linolenic acid，linoleic acid，lactate，D-glucose，glycerol，hexadecanoic acid，glycocholate and β-Hydroxybutanoate.The content of lactate in heat stress status increased，the other metabolites decreased.Functional pathway analysis revealed that heat stress aggravated the state of energy negative balance in cow，and the lactating dairy cow blood metabolically responded to heat stress through increasing β-oxidation of fatty acid and glycerol metabolic and decreasing in glycolosis activity accompanied with liver dysfuction.The study may provide a strong evidence for further researching the physiological mechanism of heat stress cow.

Key words: heat stress, dariy cows, metabonomics, GC-MS, blood metabolites

中图分类号:

S815.4

郭延生,贾启鹏,陶金忠. 基于GC-MS策略的奶牛热应激血液代谢组学研究[J]. 畜牧兽医学报, 2015, 46(8): 1356-1362.

GUO Yan-sheng，JIA Qi-peng，TAO Jin-zhong. Blood Metabolomic Studies of Heat Stress Cow with GC-MS[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(8): 1356-1362.

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