蛋白质组学技术解析急性免疫应激影响肉仔鸡肉品质的机理

doi:10.11843/j.issn.0366-6964.2021.08.007

摘要/Abstract

摘要： 旨在探讨免疫应激对肉仔鸡胸肌肉品质的影响机理。采用非标定量蛋白质组学技术研究免疫应激对AA肉仔鸡胸肌肉品质和蛋白质组学变化的影响。本试验选取120只1日龄健康AA肉仔公鸡，随机分为2个处理，分别为对照组和免疫应激处理组，每个处理6个重复，每个重复10只鸡。分别在36、38、40日龄时，于腹腔注射1 mL生理盐水（对照组）或5.0 mg·kg^-1体重的LPS溶液（免疫应激组）。42日龄时，每重复随机抽取2只鸡，采集胸肌肌肉组织测定滴水损失、蒸煮损失、肌苷酸含量和pH，及蛋白质组的定性、定量分析。结果显示：1）与对照组相比，免疫应激显著增加了肉鸡胸肌的滴水损失和蒸煮损失（P<0.05），显著降低了胸肌内肌苷酸（IMP）含量（P<0.05）；免疫应激对肉鸡胸肌宰后静置24和48 h的pH影响不显著（P>0.05）；免疫应激组肉鸡胸肌肌纤维的横截面周长和面积均极显著增加（P<0.01）。2）定性蛋白质组学分析结果显示，对照组特异表达蛋白质功能富集到了细胞呼吸、有氧呼吸、己糖代谢与生物合成过程、单糖生物合成过程、葡萄糖代谢过程和糖异生生物学过程；而免疫应激组特异表达蛋白质富集到了紧密连接代谢通路和大分子复合物分解、蛋白质分解与解聚、核酸合成和代谢、糖基化合物合成与代谢和含碱基小分子物质代谢生物学过程。3）定量蛋白质组学分析表明，免疫应激组和对照组有21个蛋白质发生差异化表达，其中15个蛋白质在免疫应激组上调表达，6个蛋白质在免疫应激组下调表达；免疫应激组上调表达的蛋白包括参与碳水化合物代谢和能量产生相关蛋白（AMPD1、GPD1L2、LDHA、UQCRC2）、肌肉收缩相关蛋白（MYH1C、MYH1A、CASQ2、TMEM38A）和参与免疫反应保护细胞免受损伤相关蛋白（HMGB1、PPP1R1A、LOC101747971、PRDX1、ABCF2）；免疫应激组下调表达的蛋白有HBBA、CS、ATP5A1WL、HSP90B1、ABCB6和SYNJ2。综上所述，LPS诱导的免疫应激通过改变肉鸡肌肉与能量代谢和肌肉收缩相关蛋白质的表达，从而增加了肌纤维的面积，并降低了肌细胞持水力，导致肉品质下降。

关键词: 肉鸡, 免疫应激, 肉品质, 蛋白质组学, 胸大肌

Abstract: The aim of this study was to investigate molecular mechanism of the effects of immunological stress on pectoralis major (PM) meat quality of AA broilers. The effects of immunological stress on the proteomic changes of PM of AA broilers were studied by using the label-free LC-MS proteomics technique. One hundred and twenty healthy one-day-old male AA broilers were randomly divided into control group and immunological stress group challenged with Escherichia coli lipopolysaccharide (LPS), with 6 replicates per treatment and 10 chickens per replicate. At the age of 36, 38 and 40 days, 1 mL of sterile saline for control group or LPS dissolved in saline at an approximate dose of 5.0 mg·kg^-1 of bodyweight for immunological stress group was injected intraperitoneally, respectively. At the age of 42 days, 2 broilers were selected from each replicate randomly, and the tissue samples of PM were collected for measuring the drip loss, cooking loss, inosine monophosphate (IMP) content and pH, and the qualitative and quantitative analysis of proteomics were performed. The results showed that:1) Compared with the control group, immunological stress significantly increased the drip loss and cooking loss of PM of broilers (P<0.05), and significantly reduced the content of IMP (P<0.05); Immunological stress had no significant effect on pH of PM of broilers slaughtered after 24 and 48 h (P>0.05); The cross sectional perimeter and area of PM muscle fiber in immunological stress group were significantly increased (P<0.01). 2) The results of qualitative proteomics analysis showed, in the control group, the specially expressed proteins were enriched in cell respiration, aerobic respiration, hexose metabolism and biosynthesis, monosaccharide biosynthesis, glucose metabolism and gluconeogenesis; While, in the immunological stress group, the specially expressed proteins were enriched in the tight junction metabolic pathway, the biological processes included macromolecular complex decomposition and protein metabolism, decomposition and depolymerization of protein, synthesis and metabolism of nucleic acid and glycosyl compounds, metabolism of nucleobase-containing small molecule. 3) Quantitative proteomics analysis showed that:21 proteins were differentially expressed in the immunological stress group and the control group, 15 of which were up-regulated and 6 were down-regulated in the immunological stress group; The up-regulated proteins in the immunological stress group included the proteins involved in carbohydrate metabolism and energy production (AMPD1, GPD1L2, LDHA, UQCRC2), muscle contraction related proteins (MYH1C, MYH1A, CASQ2, TMEM38A) and proteins involved in immunological response to protect cells from damage (HMGB1, PPP1R1A, LOC101747971, PRDX1,1ABCF2); The down-regulated protein in the immunological stress group were HBBA, CS, ATP5A1WL, HSP90B1, ABCB6 and SYNJ2. The results indicated that immunological stress induced by LPS increased the muscle fiber area, reduced the cell hydraulic holding capacity of muscle cells, and resulted in the poor meat quality by changing the expression of proteins related to the energy metabolism and muscle contraction in PM.

Key words: broiler chickens, immunological stress, meat quality, proteomics, pectoralis major muscle

中图分类号:

S831.2

张安荣, 吴正可, 陈志敏, 常文环, 蔡辉益, 刘国华, 郑爱娟. 蛋白质组学技术解析急性免疫应激影响肉仔鸡肉品质的机理[J]. 畜牧兽医学报, 2021, 52(8): 2138-2150.

ZHANG Anrong, WU Zhengke, CHEN Zhimin, CHANG Wenhuan, CAI Huiyi, LIU Guohua, ZHENG Aijuan. Unraveling Molecular Mechanism of Acute Immunological Stress Affecting Meat Quality of Broiler Chickens by Proteomics Analysis[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(8): 2138-2150.

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