Chinese Journal of Animal Nutrition >

2024 , Vol. 36 >Issue 8: 5308 - 5318

DOI: https://doi.org/10.12418/CJAN2024.451

RESEARCH PAPER

Effects of Yeast Peptides on Growth Performance of Meat Rabbits, and Serum Cytokines and Liver Antioxidant Capacity of Lipopolysaccharide-Stressed Meat Rabbits

  • FAN Jiaqi , 1 ,
  • HAN Wenxiao 2, 3 ,
  • FAN Huimin 2, 3 ,
  • CHEN Baojiang 1 ,
  • WANG Hong 4 ,
  • ZHI Yongwei 5 ,
  • CUI Qiujia 6 ,
  • CHEN Saijuan , 1, 2, 3, *
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  • 1 College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, China
  • 2 Mountainous Area Research Institute of Hebei Province, Hebei Agricultural University, Baoding 071001, China
  • 3 Agricultural Technology Innovation Centre in Mountainous Areas of Hebei Province, Baoding 071001, China
  • 4 Beijing Enhalor Biotechnology Co., Ltd., Beijing 101109, China
  • 5 Inner Mongolia Daerduo Rabbit Industry Co., Ltd., Ordos 014300, China
  • 6 Zaoqiang Animal Husbandry Station, Hengshui 053100, China
*associate professor, E-mail:

Received date: 2024-02-04

  Online published: 2024-08-12

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Abstract

The purpose of this study was to investigate the effects of yeast peptide on the growth performance of meat rabbits and serum cytokines and liver antioxidant capacity of lipopolysaccharide-stressed meat rabbits. The trial was divided into two phases. Phase 1: a total of 160 healthy 35-day-old weaned Hyla line rabbits were selected with an initial body weight of (0.96±0.06) kg, and randomly divided into 4 groups (half male and half female), with 8 replicates per group and 5 rabbits per replicate. Rabbits in control group (CON group) and lipopolysaccharide group (LPS group) fed a basal diet, and those in yeast peptide group (YP group) and yeast peptide remission group (L-YP group) fed the basal diet supplemented with 100 mg/kg yeast peptide. The feeding experiment lasted for 35 days, including 7 days of pre-trial period and 28 days of experimental period. Data such as body weight and feed intake were recorded and growth performance indexes of meat rabbits during the phase 1 were calculated. Phase 2: at the end of phase 1, rabbits in the LPS group and L-YP group were intramuscularily injected with 200 μg/kg BW LPS, and rabbits in the CON group and YP group were injected with the same amount of normal saline. The samples were collected 4 h after injection with LPS or normal saline, and serum cytokine content, liver antioxidant indexes and mRNA relative expression levels of antioxidant genes were measured. The results showed as follows: 1) adding yeast peptide significantly increased the average daily gain and significantly decreased the feed to gain ratio of meat rabbits (P<0.05), but had no significant effects on the average daily feed intake, diarrhea rate and mortality rate (P>0.05). 2) The contents of serum interleukin-6 (IL-6), interleukin-8 (IL-8) and reactive oxygen species (ROS) were significantly decreased by adding yeast peptide (P<0.05); lipopolysaccharide stress significantly increased the contents of serum tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), IL-6, IL-8, transforming growth factor-β (TGF-β) and ROS of meat rabbits (P<0.05); serum TNF-α, IL-6, IL-10 and TGF-β contents had significant interactions between yeast peptide and lipopolysaccharide (P<0.05). Under lipopolysaccharide stress, adding yeast peptide significantly decreased the serum TNF-α and IL-6 contents of lipopolysaccharide-stressed meat rabbits (P<0.05). 3) Adding yeast peptide significantly increased the liver superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activities and total antioxidant capacity (T-AOC) of meat rabbits (P<0.05), and significantly decreased the liver malondialdehyde (MDA) content (P<0.05); lipopolysaccharide stress significantly decreased the liver T-AOC and GSH-Px activity (P<0.05); SOD activity and SOD mRNA relative expression level in liver had a significant interaction between yeast peptide and lipopolysaccharide (P<0.05). Under lipopolysaccharide stress, adding yeast peptide significantly increased the liver SOD activity and SOD mRNA relative expression level of lipopolysaccharide-stressed meat rabbits (P<0.05). In summary, adding 100 mg/kg yeast peptide to the diet can improve the growth performance of meat rabbits, and effectively relieve the immune stress response of meat rabbits induced by lipopolysaccharide by reducing the level of inflammation and improving the antioxidant capacity.

Key words： yeast peptide; meat rabbits; lipopolysaccharide; inflammation; antioxidant capacity

Cite this article

FAN Jiaqi , HAN Wenxiao , FAN Huimin , CHEN Baojiang , WANG Hong , ZHI Yongwei , CUI Qiujia , CHEN Saijuan . Effects of Yeast Peptides on Growth Performance of Meat Rabbits, and Serum Cytokines and Liver Antioxidant Capacity of Lipopolysaccharide-Stressed Meat Rabbits[J]. Chinese Journal of Animal Nutrition, 2024 , 36(8) : 5308 -5318 . DOI: 10.12418/CJAN2024.451

兔肉味道鲜美、营养价值丰富,近年来,国际市场对兔肉的需求不断升温,我国兔肉出口量逐年增长,冷冻兔肉出口量占到了世界冷冻兔肉出口总量的70%左右。当前高度集约化的饲养模式,使肉兔的发病率大大增加[1],肉兔断奶前后肠道尚未发育完全,免疫力低,换料、环境、温度、细菌等刺激极易使断奶仔兔处于应激状态,影响机体正常的代谢功能,导致肉兔出现食欲减退、腹泻甚至死亡等情况,不利于其健康生长[2]。抗菌肽具有提高畜禽成活率、促进生长发育、改善肠道健康、提高机体免疫力的作用,已广泛应用于猪[3]、鸡[4]、牛[5]、羊[6]等生产实践中。酵母肽(yeast peptide,YP)是由酿酒酵母产生的一种抗菌肽。研究证实,饲粮中添加酵母肽可显著提高仔猪的生长性能[7]。本课题组前期研究了酵母肽对肉兔生长性能、免疫性能、抗氧化能力和肠道健康的影响,结果显示酵母肽能提高肉兔的生长性能和免疫功能,改善肠道健康,且以100 mg/kg酵母肽的作用效果较好[8]。脂多糖(lipopolysaccharide,LPS)是革兰氏阴性菌细胞壁的主要成分,LPS刺激会导致机体内环境稳态遭到破坏,如加剧炎症水平、损伤肠黏膜形态和屏障功能,降低机体免疫力和抗氧化能力等[9],从而导致动物产生严重的健康问题。LPS诱导的动物应激模型操作简单、容易重复,是目前动物营养学公认的造模方法。基于本课题组先前的研究结果,本试验在饲粮中添加100 mg/kg酵母肽,拟探究酵母肽对肉兔生长性能的影响,以及对LPS应激肉兔血清细胞因子和肝脏抗氧化指标的影响,以阐明酵母肽对肉兔健康的调节作用。

1 材料与方法

1.1 试验材料

试验所用酵母肽产品的酵母肽含量≥5 000 mg/kg;LPS来源于大肠杆菌O55∶B5,购自Sigma公司,货号为L2880。基础饲粮依据《肉兔营养需要量》(NY/T 4049—2021)[10]进行配制,其组成及营养水平见表1。饲粮消化能参照NY/T 4049—2021计算得出,粗蛋白质、粗脂肪、粗纤维、中性洗涤纤维、酸性洗涤纤维、酸性洗涤木质素、粗灰分、钙、总磷和氨基酸含量的测定分别参照GB/T 6432—2018、GB/T 6433—2006、GB/T 6434—2022、GB/T 20806—2022、NY/T 1459—2022、GB/T 20805—2006、GB/T 6438—2007、GB/T 6436—2018、GB/T 6437—2018和GB/T 18246—2019。
表1 基础饲粮组成及营养水平(风干基础)

Table 1 Composition and nutrient levels of the basal diet (air-dry basis) %

项目Items 含量Content
原料Ingredients
玉米Corn 9.00
小麦麸Wheat bran 20.00
豆粕Soybean meal 2.00
次粉Wheat middling 8.00
葵花饼Sunflower cake 11.00
麦芽根Malt root 12.00
花生壳Peanut hulls 22.00
艾叶粉Artemsia argyi powder 5.00
石粉Limestone 1.50
食盐NaCl 0.50
稻壳粉Rice husk powder 7.00
磷酸氢钙CaHPO4 0.55
L-赖氨酸盐酸盐L-lysine hydrochloride 0.35
DL-蛋氨酸DL-Met 0.10
预混料Premix1) 1.00
合计Total 100.00
营养水平Nutrient levels2)
消化能DE/(MJ/kg) 10.15
粗蛋白质CP 16.79
赖氨酸Lys 0.85
蛋氨酸+胱氨酸Met+Cys 0.57
粗脂肪EE 3.27
粗纤维CF 15.32
中性洗涤纤维NDF 41.56
酸性洗涤纤维ADF 24.83
酸性洗涤木质素ADL 5.71
粗灰分Ash 11.27
钙Ca 1.34
总磷TP 0.69

1)预混料为每千克饲粮提供 The premix provided the following per kg of the diet:Fe 70 mg,Cu 20 mg,Zn 70 mg,Mn 10 mg,Co 0.15 mg,I 0.2 mg,Se 0.25 mg,VE 50 mg,VK 2 mg,VB1 2 mg,VB2 6 mg,VB5 50 mg,VB6 2 mg,VB12 0.02 mg,VB3 50 mg,烟酸 niacin 20 mg,泛酸 pantothenic acid 12.5 mg,VA 10 000 IU,VD 900 IU,胆碱 choline 1 000 mg,生物素 biotin 0.2 mg。

2)营养水平中消化能为计算值,其他为实测值。In nutrient levels, DE was a calculated value, while the others were measured values.

1.2 试验设计和饲养管理

试验分2个阶段进行。第1阶段:选取35日龄的断奶健康伊拉配套系肉兔160只,初始体重为(0.96±0.06) kg,随机分为4组,每组的80只肉兔中公母各占1/2,将每组的80只肉兔分为8个重复,每个重复5只,其中对照组(CON组)和LPS组饲喂基础饲粮,酵母肽组(YP组)和酵母肽缓解组(L-YP组)饲喂添加100 mg/kg酵母肽的基础饲粮。第1阶段饲养试验共进行35 d,其中预试期7 d,正试期28 d。记录并计算第1阶段试验期间肉兔的生长性能数据。第2阶段:采用2×2双因素试验设计,在第1阶段试验结束时,参照刘栩渤等[11]的方法,LPS组和L-YP组试验兔大腿肌肉注射200 μg/kg BW脂多糖,CON组和YP组试验兔则注射等量生理盐水,4 h后从每组每个重复选择1只(每组8只)试验兔采样。
试验期间试验兔单笼(长×宽×高:50 cm×34 cm×33 cm)饲养(每5笼合并为1个重复),自由采食和饮水,并按照常规流程对试验兔进行免疫。

1.3 测定指标与方法

1.3.1 生长性能测定

试验兔分别于第1阶段正试期的第1天和第28天空腹称重,记录为初始体重和终末体重,并记录第1~28天各组的采食量,根据记录的数据计算此阶段的平均日增重(ADG)、平均日采食量(ADFI)和料重比(F/G);记录第1阶段正试期内试验兔的腹泻和死亡情况,计算腹泻率和死亡率,计算公式为:
腹泻率(%)=(腹泻兔只数/试验兔总只数)×100;
死亡率(%)=(死亡兔只数/试验兔总只数)×100。

1.3.2 血清细胞因子含量测定

试验兔注射LPS或生理盐水4 h后采集血液。经耳缘静脉采血5 mL,将血液样品静置30 min后,以630×g离心10 min,取上清液,移入2 mL的EP管中,保存在-80 ℃冰箱中。采用上海酶联生物科技有限公司的酶联免疫吸附测定(ELISA)试剂盒测定血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)、白细胞介素-10(IL-10)、转化生长因子-β(TGF-β)、活性氧(ROS)和溶菌酶(LZM)含量,具体操作步骤严格按照说明书指示进行。

1.3.3 肝脏抗氧化指标测定

采血后,将试验兔安乐死(静脉注射100 mg/kg BW的戊巴比妥钠),取适当肝脏组织样本放于冻存管中,-80 ℃保存备测。分析天平称量适量肝脏样品于2 mL的EP管中,按照1∶9的质量体积比加入生理盐水,低温研磨,制成10%肝脏组织匀浆。采用上海酶联生物科技有限公司的ELISA试剂盒测定肝脏中总抗氧化能力(T-AOC)以及谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,具体操作步骤严格按照说明书指示进行。

1.3.4 肝脏抗氧化相关基因mRNA表达的检测

采用实时荧光定量PCR法测定肝脏组织中SOD和GSH-Px的mRNA相对表达量,实时荧光定量PCR引物序列如表2所示。使用TRIzol试剂提取肝脏组织的总RNA,采用分光光度法检测提取的RNA的浓度,并用PrimeScript RT Reagent Kit合成cDNA。在MA-6000型实时荧光定量PCR仪上进行PCR,反应体系总体积为20 μL,由10 μL 2×SYBR real-time PCR premixture、0.8 μL正向和反向引物混合物、1 μL cDNA和8.2 μL RNase Free dH2O组成。实时荧光定量PCR热循环条件为:95 ℃热循环5 min,95 ℃热循环15 s,60 ℃热循环30 s,共循环40次。以β-肌动蛋白(β-actin)为内参基因,重复3次,用2-△△Ct法计算目的基因的mRNA相对表达量。
表2 实时荧光定量PCR引物序列

Table 2 Primer sequences used for RT-qPCR

基因名称
Gene names		 引物序列
Primer sequence (5'—3')		 产物长度
Product length/bp		 序列号
Accession number
超氧化物歧化酶
SOD		 F:ACCTGGGTAATGTGACTGCA
R:CCCGGTCTTTGTACTCTCGT		 167 NC_067387.1
谷胱甘肽过氧化物酶
GSH-Px		 F:AAGCTCATCACCTGGTCTCC
R:TCAGGCTCGATGTCAATGGT		 131 NC_000003.12
β-肌动蛋白
β-actin		 F:CCTGGCACCCAGCACAAT
R:GCTGATCCACATCTGCTGGAA		 118 NW_026259299.1

1.4 数据处理与分析

生长性能数据以重复为统计单位,结果用平均值±标准差表示,采用SPSS 22.0软件进行单因素方差分析,并采用Duncan氏法进行组间多重比较。LPS应激肉兔的血清细胞因子含量、肝脏抗氧化指标与抗氧化基因mRNA相对表达量结果用平均值和均值标准误表示,通过SPSS 22.0软件的一般线性模型进行2×2析因处理分析,统计模型涉及酵母肽(添加0或100 mg/kg)和LPS(注射生理盐水或LPS)以及它们之间的交互效应,当存在显著的交互效应时,用Duncan氏法进行组间多重比较。以P<0.05表示差异显著,0.05≤P<0.10表示差异有显著趋势。

2 结果与分析

2.1 酵母肽对LPS应激前肉兔生长性能的影响

由表3可知,与基础饲粮组(包含CON组和LPS组)相比,酵母肽组(包含YP组和L-YP组)肉兔的ADG显著增加(P<0.05),F/G显著降低(P<0.05),ADFI、腹泻率和死亡率无显著变化(P>0.05)。
表3 酵母肽对肉兔生长性能的影响

Table 3 Effects of yeast peptide on growth performance of meat rabbits (n=16)

项目
Items		 基础饲粮组
Basal diet group		 酵母肽组
Yeast peptide group		 P值
P-value
平均日增重ADG/g 37.33±1.51b 40.61±2.04a 0.003
平均日采食量ADFI/g 151.89±5.75 156.85±3.61 0.158
料重比F/G 4.07±0.17a 3.87±0.17b 0.032
腹泻率Diarrhea rate/% 6.25 2.50 0.246
死亡率Mortality rate/% 8.75 3.75 0.191

同行数据肩标不同小写字母表示差异显著(P<0.05),无字母或有相同字母表示差异不显著(P>0.05)。

In the same row, values with different small letter superscripts mean significant difference (P<0.05), while with no letter or the same letter superscripts mean no significant difference (P>0.05).

2.2 酵母肽对LPS应激肉兔血清细胞因子含量的影响

由表4可知,饲粮中添加酵母肽显著提高了肉兔血清中TGF-β含量(P<0.05),显著降低了IL-6、IL-8和ROS含量(P<0.05),有升高IL-10含量的趋势(P=0.086),有降低IL-1β含量的趋势(P=0.086);LPS刺激显著提高了肉兔血清中促炎细胞因子TNF-α、IL-1β、IL-6、IL-8、TGF-β和ROS含量(P<0.05)。酵母肽和LPS对肉兔血清中TNF-α、IL-6、IL-10和TGF-β含量有显著的交互效应(P<0.05)。在LPS刺激条件下,饲粮中添加酵母肽可以显著降低血清中TNF-α、IL-6含量(P<0.05),说明酵母肽在一定程度上可缓解LPS应激肉兔的炎症反应。
表4 酵母肽对LPS应激肉兔血清细胞因子含量的影响

Table 4 Effects of yeast peptide on serum cytokine contents of LPS-stressed meat rabbits (n=8)

项目
Items		 肿瘤坏死
因子-α
TNF-α/
(pg/mL)		 白细胞
介素-1β
IL-1β/
(pg/mL)		 白细胞
介素-6
IL-6/
(pg/mL)		 白细胞
介素-8
IL-8/
(pg/mL)		 白细胞
介素-10
IL-10/
(pg/mL)		 转化生长
因子-β
TGF-β/
(pg/mL)		 活性氧
ROS/
(pg/mL)		 溶菌酶
LZM/
(ng/mL)

LPS(-)		 YP(-) 471.46b 134.71 235.92b 249.32 100.77b 4 265.31b 573.88 1 633.36
YP(+) 581.08b 120.84 231.43b 143.67 120.65a 6 426.20a 461.05 1 668.94

LPS(+)		 YP(-) 754.06a 166.80 276.37a 308.50 111.34ab 6 566.87a 661.71 1 780.00
YP(+) 530.15b 143.90 224.08b 218.95 108.57ab 6 407.50a 604.80 1 664.10
SEM 36.24 10.29 6.91 26.79 4.79 232.19 31.36 71.91
主效应Main effects

YP		 YP(-) 612.76 150.76 256.15a 278.91a 106.06 5 416.09b 617.80a 1 706.68
YP(+) 555.62 132.38 227.76b 181.31b 114.61 6 416.88a 532.93b 1 666.54

LPS		 LPS(-) 526.27b 127.78b 233.68b 196.50b 110.71 5 345.78b 517.47b 1 651.15
LPS(+) 642.11a 155.36a 250.23a 263.73a 109.96 6 487.19a 633.26a 1 722.07
P值P-value
YP 0.128 0.089 0.001 0.002 0.086 <0.001 0.011 0.581
LPS 0.004 0.014 0.029 0.021 0.876 <0.001 0.001 0.333
YP×LPS <0.001 0.666 0.003 0.767 0.025 <0.001 0.380 0.301

LPS:脂多糖;YP:酵母肽;LPS(-):脂多糖刺激;LPS(+):无脂多糖刺激;YP(-):未添加酵母肽;YP(+):添加酵母肽;YP×LPS:脂多糖与酵母肽的交互作用。同列数据肩标不同小写字母表示差异显著(P<0.05),无字母或有相同字母表示差异不显著(P>0.05)。下表同。

LPS: lipopolysaccharide; YP: yeast peptide; LPS(-): stimulation with lipopolysaccharide; LPS(+): non-stimulation with lipopolysaccharide; YP(-): no yeast peptide was added; YP(+): yeast peptide was added; YP×LPS: the interaction of lipopolysaccharide and yeast peptide. In the same column, values with different small letter superscripts mean significant difference (P<0.05), while with no letter or the same letter superscripts mean no significant difference (P>0.05). The same as below.

2.3 酵母肽对LPS应激肉兔肝脏抗氧化能力的影响

由表5可知,饲粮中添加酵母肽显著提高了肉兔肝脏中SOD、GSH-Px活性和T-AOC(P<0.05),显著降低了肝脏中MDA含量(P<0.05);LPS刺激显著降低了肉兔肝脏中T-AOC和GSH-Px活性(P<0.05)。酵母肽和LPS对肉兔肝脏中SOD活性有显著的交互效应(P<0.05)。在LPS刺激条件下,饲粮中添加酵母肽可以显著提高LPS应激肉兔肝脏中SOD活性(P<0.05)。
表5 酵母肽对LPS应激肉兔肝脏抗氧化指标的影响

Table 5 Effects of yeast peptide on antioxidant indexes in liver of LPS-stressed meat rabbits (n=8)

项目
Items		 总抗氧化能力
T-AOC/
(U/mg prot)		 超氧化物歧化酶
SOD/
(U/mg prot)		 谷胱甘肽过氧化物酶
GSH-Px/
(U/mg prot)		 丙二醛
MDA/
(nmol/mg prot)

LPS(-)		 YP(-) 25.18 203.67b 24.15 8.14
YP(+) 32.09 249.76b 29.44 6.58

LPS(+)		 YP(-) 20.38 115.05c 20.85 8.52
YP(+) 27.58 368.23a 26.07 7.26
SEM 1.91 28.91 1.55 0.47
主效应Main effects

YP		 YP(-) 22.79b 159.36b 22.50b 8.33a
YP(+) 29.84a 309.00a 27.76a 6.93b

LPS		 LPS(-) 28.64a 226.72 26.80a 7.36
LPS(+) 23.99b 241.65 23.47b 7.90
P值P-value
YP 0.001 <0.001 0.002 0.006
LPS 0.022 0.610 0.041 0.266
YP×LPS 0.939 0.001 0.981 0.748
由表6可知,单纯地添加酵母肽或进行LPS刺激均未对肉兔肝脏中SOD和GSH-Px的mRNA相对表达量产生显著影响(P<0.05),但二者对肝脏中SOD的mRNA相对表达量有显著的交互效应(P<0.05),对GSH-Px的mRNA相对表达量有产生交互效应的趋势(P=0.079)。在LPS刺激条件下,饲粮中添加酵母肽可以显著提高LPS应激肉兔肝脏中SOD的mRNA相对表达量(P<0.05)。
表6 酵母肽对LPS应激肉兔肝脏抗氧化基因mRNA相对表达量的影响

Table 6 Effects of yeast peptide on mRNA relative expression levels of antioxidant genes in liver of LPS-stressed meat rabbits (n=5)

项目
Items		 超氧化物歧化酶
SOD		 谷胱甘肽过氧化物酶
GSH-Px

LPS(-)		 YP(-) 1.07ab 1.01
YP(+) 0.95ab 0.95

LPS(+)		 YP(-) 0.76b 0.76
YP(+) 1.12a 1.11
SEM 0.10 0.10
主效应Main effects

YP		 YP(-) 0.92 0.89
YP(+) 1.03 1.04

LPS		 LPS(-) 1.01 0.98
LPS(+) 0.94 0.94
P值P-value
YP 0.280 0.197
LPS 0.514 0.697
YP×LPS 0.049 0.079

3 讨论

3.1 酵母肽对肉兔生长性能的影响

酵母肽是一种微生物源抗菌肽,通过诱导酿酒酵母产生,作为一种具有多种生物活性(如抗炎[12]、抗应激[13]、促进生长和免疫调节[14]等)的营养添加剂已在畜牧养殖行业广泛应用。Shastri等[15]研究发现,饲粮中添加抗菌肽(100~200 mg/kg)可显著提高34周龄蛋鸡的体重,并提高生产性能。李波等[16]研究表明,抗菌肽可以提高断奶仔猪的ADG,降低F/G,并通过提高血清中免疫球蛋白A(IgA)和免疫球蛋白G(IgG)含量提高免疫力。Choi等[17]研究表明,90 mg/kg抗菌肽-A3能够改善肉鸡的生长性能,提高干物质、总能和粗蛋白质的表观代谢率。本课题组来宁洁[8]的研究发现,饲粮中添加100、200和300 mg/kg酵母肽均能提高肉兔的生长性能和免疫功能,降低腹泻率和死亡率,改善肠道组织结构,调节肠道菌群组成,其中100 mg/kg酵母肽对肉兔的改善作用要优于其他剂量,所以本研究在其基础上选用100 mg/kg酵母肽进行试验。本试验先进行了28 d的饲养试验(LPS应激前),结果显示,添加100 mg/kg酵母肽可提高肉兔的ADFI和ADG,降低F/G,与本课题组先前研究所得结果[8]一致。这一方面可能与酵母肽的分子质量有关,酵母肽的分子质量只有1.9 kDa,有助于在水介质中有更高的溶解度,使其更高效地被消化吸收以及进入细胞发挥作用[18];另一方面可能与抗菌肽能够提高免疫力、促进肠道发育和改善动物健康状态有关。抗菌肽可以参与分化、增殖、成熟和凋亡等细胞代谢过程[19],提高肠上皮细胞紧密连接蛋白的表达,改善肠道发育,并且抑制肠道内的有害菌,整体提升动物健康水平,提高对营养物质的消化吸收,进而提高生长性能[20]。

3.2 酵母肽对LPS应激肉兔血清细胞因子含量的影响

LPS诱导的动物应激模型因其操作简单、可重复性高而被人们广泛应用[21]。LPS是革兰氏阴性菌细胞壁上的组成成分,与Toll样受体4(TLR4)结合后,激活下游信号通路,使炎症通路相关调控因子过度表达,刺激下游TNF-α、IL-6、IL-8等炎性细胞因子大量产生[22],使动物处于免疫应激状态[23]。本研究发现,LPS应激显著提高了肉兔血清中促炎细胞因子TNF-α、IL-1β、IL-6和IL-8的含量,说明LPS应激引起了免疫系统的过度激活,产生了炎症;而在饲粮中添加酵母肽则显著降低了应激状态下肉兔血清中TNF-α和IL-6含量,缓解LPS诱导的应激反应。有研究表明,抗菌肽能够通过抑制炎症信号通路中的关键因子核因子-κB(NF-κB)的活性减少TNF-α的表达[24],还可以抑制炎性单核细胞产生TNF-α、IL-1β和IL-6[25],甚至直接或间接参与炎性因子蛋白的翻译、稳定或加工,破坏其结构和功能[26]。IL-10是一种可以由所有免疫细胞产生的细胞因子,能够抑制T细胞、NK细胞和巨噬细胞合成和分泌炎性细胞因子,起到抗炎作用[27]。酵母肽有增加肉兔血清中IL-10含量的趋势,说明酵母肽在一定程度上缓解了肉兔的炎症水平。TGF-β可以参与细胞增殖分化和修复受损组织[28]。尉心田等[29]研究发现,LPS刺激后,仔猪肝脏TGF-β1/Smads信号通路被激活,参与肝脏的损伤修复。本试验结果与之一致,酵母肽和LPS应激均会显著提高肉兔血清TGF-β含量,前者是对机体的保护和促进作用,后者则体现了受损组织的修复情况。LPS刺激导致肉兔血清中ROS含量显著增加,机体中过量的ROS会导致细胞功能障碍,影响骨骼肌细胞的发育和再生[30]。王瑛等[31]研究发现,在荷斯坦公牛的基础饲粮中添加8 g/d抗菌肽,可以显著降低血清中ROS含量。本试验中,饲粮中添加酵母肽显著降低了肉兔血清中ROS含量,说明酵母肽在一定程度上缓解了LPS刺激带来的过氧化反应,维持了机体内环境的稳态。

3.3 酵母肽对LPS应激肉兔肝脏抗氧化能力的影响

动物的抗氧化能力与其健康密切相关。通常情况下,动物体内的氧化和抗氧化水平保持相对平衡。然而,一旦这种平衡被破坏,自由基的过量产生可能导致氧化应激和细胞损伤[32]。GSH-Px和SOD是主要分布在细胞器膜上的抗氧化酶,由LPS诱导所产生的过量自由基通过攻击细胞及细胞器的膜结构,导致细胞膜结构受损,使得抗氧化酶失去发挥活性的载体,最终降低抗氧化酶的活性[33]。T-AOC可反映机体整体的抗氧化水平,MDA含量增加,表明细胞脂质过氧化程度加深。抗菌肽作为抗氧化剂在动物生产中已有较多应用,吕志鹏[34]研究表明,不同剂量的抗菌肽均能够增加早期断奶仔猪血清中GSH-Px和SOD活性,显著降低血清中MDA含量,并且呈现出剂量依赖性。胡文举等[35]研究发现,饲粮中添加200 mg/kg抗菌肽可显著增加固始鸡血清中T-AOC,提高血清中GSH-Px活性。在本试验中,酵母肽可以使肉兔肝脏中GSH-Px、SOD活性和T-AOC增加,肝脏中MDA含量下降,而LPS刺激降低了肝脏中GSH-Px活性和T-AOC。张海文等[36]研究发现,仔猪腹腔注射0.5 mg/kg BW的抗菌肽后,有效缓解了LPS诱导的氧化损伤,肝脏的抗氧化酶活性得到有效的恢复。本试验结果与之类似,这说明酵母肽提高了肉兔肝脏的抗氧化能力,减轻了LPS诱导的氧化损伤。在基因表达方面,也显示了相同的结果。

4 结论

综上所述,饲粮中添加100 mg/kg酵母肽能够提高肉兔的生长性能,并能通过降低机体的炎症水平、提高肝脏抗氧化能力有效缓解LPS诱导的肉兔免疫应激反应。

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