Chinese Journal of Animal Nutrition >

2024 , Vol. 36 >Issue 5: 3015 - 3023

DOI: https://doi.org/10.12418/CJAN2024.259

RESEARCH PAPER

Study on Alleviation of Dietary Betaine on Corticosterone Induced Small Intestinal Inflammation of Laying Hens

  • DU Xin , 1 ,
  • LI Ding 1 ,
  • WANG Shengchen 1 ,
  • CUI Xiaoyan 1 ,
  • LI Tingting 1 ,
  • ZHANG Liyang 2 ,
  • LUO Xugang , 1, * ,
  • HU Yun , 1, *
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  • 1 College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China
  • 2 Mineral Nutrition Research Division, Institute of Animal Science, Chinese Academy of Agricultural Science, Beijing 100193, China
* LUO Xugang, professor, E-mail: ;
HU Yun, associate professer, E-mail:

Received date: 2022-12-05

  Online published: 2024-05-15

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Abstract

This experiment was conducted to investigate the alleviation of dietary betaine on corticosterone induced small intestinal inflammation of laying hens. Seventy-two 130-day-old laying hens were divided into 3 groups with 8 replicates per group and 3 laying hens per replicate according a completely randomized design. Laying hens in the control and corticosterone groups were fed a basal diet, and others in betaine+corticosterone group were fed the basal diet+0.1% betaine. At 28 days of the experiment, the laying hens in corticosterone and betaine+corticosterone groups were received subcutaneous injections of 4.0 mg/kg BW corticosterone, and others in the control group were received subcutaneous injection of an equal amount of physiological saline, continuous injection for 7 days. The experiment lasted for 35 days. The results showed as follows: 1) compared with the control group, the content of interleukin-1β (IL-1β) in duodenum, jejunum and ileum of corticosterone group was significantly increased (P<0.05); compared with the corticosterone group, the content of IL-1β in duodenum, jejunum and ileum of betaine+corticosterone group was significantly decreased (P<0.05). 2) Compared with the control group, the mRNA expression levels of Toll-like receptor 4 (TLR4), interleukin-8 (IL-8) and nuclear factors-κB p65 (NF-κB p65) in duodenum of corticosterone group were significantly increased (P<0.05); compared with the corticosterone group, the mRNA expression levels of TLR4, IL-8 and NF-κB p65 in duodenum of betaine+corticosterone group were significantly decreased (P<0.05). 3) Compared with the control group, the mRNA expression levels of IL-1β and NF-κB p65 in jejunum of corticosterone group were significantly increased (P<0.05); compared with the corticosterone group, the mRNA expression level of IL-1β in jejunum of betaine+corticosterone group was significantly decreased (P<0.05), and the mRNA expression level of interleukin-10 (IL-10) in jejunum of betaine+corticosterone group was significantly increased (P<0.05). 4) Compared with the control group, the mRNA expression levels of IL-1β, IL-8 and NF-κB p65 in ileum of corticosterone group were significantly increased (P<0.05); compared with the corticosterone group, the mRNA expression levels of IL-1β, IL-8 and NF-κB p65 in ileum of betaine+corticosterone group were significantly decreased (P<0.05). In conclusion, dietary betaine supplementation can alleviate the corticosterone induced small intestinal inflammation corticosterone of laying hens via reducing small intestinal IL-1β content and inflammation related genes mRNA expression levels.

Key words： betaine; corticosterone; laying hens; small intestine; inflammation

Cite this article

DU Xin , LI Ding , WANG Shengchen , CUI Xiaoyan , LI Tingting , ZHANG Liyang , LUO Xugang , HU Yun . Study on Alleviation of Dietary Betaine on Corticosterone Induced Small Intestinal Inflammation of Laying Hens[J]. Chinese Journal of Animal Nutrition, 2024 , 36(5) : 3015 -3023 . DOI: 10.12418/CJAN2024.259

近年来,随着蛋鸡养殖业向着集约化和规模化的方式快速发展,在提高养殖效率的同时,也使得鸡群遭受着各种各样的应激,如病毒感染、高饲养密度和热应激等[1]。鸡群受到应激时,肾上腺皮质会快速合成并释放糖皮质激素——皮质酮。长期的慢性应激会诱导产蛋鸡肠道发生炎症反应,进而影响产蛋鸡肠道的消化和吸收,降低其生产性能,给蛋鸡养殖业造成严重的经济损失[2]。先前的研究表明,抗生素在治疗动物肠道炎症反应中发挥重要作用[3]。然而,我国已于2020年开始全面禁止在畜禽养殖中使用各类抗生素。因此,寻找一种绿色新型饲料添加剂缓解应激诱导的产蛋鸡肠道炎症反应成为当下研究热点。
甜菜碱是一种生物碱,为甜菜加工提取蔗糖时的天然副产品,在许多微生物、海洋无脊椎动物、植物和动物中广泛分布[4]。李鼎等[5]研究表明,饲粮添加甜菜碱可通过降低产蛋鸡肝脏中白细胞介素-1β(interleukin-1β,IL-1β)mRNA表达水平,缓解皮质酮诱导的肝脏炎症反应。但是,甜菜碱是否可以改善皮质酮诱导的产蛋鸡肠道炎症反应尚不清楚。因此,本试验通过皮下注射皮质酮建立产蛋鸡应激模型,研究饲粮添加甜菜碱对皮质酮诱导的产蛋鸡小肠各段组织中炎症因子含量和炎症相关基因mRNA表达水平的影响,探讨甜菜碱对皮质酮诱导的产蛋鸡肠道炎症反应的缓解作用,为甜菜碱在蛋鸡养殖生产中的合理应用和蛋鸡产业绿色高效转型提供试验依据。

1 材料与方法

1.1 试验材料

皮质酮,货号C2505,购自Sigma公司。甜菜碱,添加形式为甜菜碱盐酸盐,购自于宜兴市天石饲料有限公司,纯度为98%,其中甜菜碱含量为75%。如皋黄鸡购自于江苏省家禽科学研究所。130~165日龄产蛋鸡玉米-豆粕型基础饲粮购自于江苏省家禽科学研究所。

1.2 试验设计

试验选用72只130日龄商品代如皋黄鸡,采用单因素完全随机试验设计,按体重和产蛋率随机分为3组,每组8个重复,每个重复3只鸡。对照组(CON组)和皮质酮组(CORT组)产蛋鸡全程饲喂不添加甜菜碱的基础饲粮,甜菜碱+皮质酮组(BET+CORT组)产蛋鸡全程饲喂基础饲粮+0.10%甜菜碱。甜菜碱添加剂量参照He等[6]的研究确定。试验至28 d时,按照先前已有的报道[7],皮质酮组和甜菜碱+皮质酮组产蛋鸡分别皮下注射4 mg/kg BW皮质酮(皮质酮先溶解于纯酒精中,再用生理盐水稀释至酒精浓度为15%),连续注射7 d,建立应激模型;对照组产蛋鸡皮下注射等量酒精浓度为15%的生理盐水。试验期为35 d。

1.3 试验饲粮与饲养管理

3组饲粮的所有养分均满足《鸡饲养标准》(NY/T 33—2004)[8]和NRC(1994)[9]中关于产蛋鸡的营养需要量。基础饲粮组成及营养水平见表1。饲养管理按照《如皋黄鸡饲养管理手册》进行,鸡只自由采食与饮水,每日光照16 h,饲粮以粉料形式供给。每天观察记录鸡舍环境温湿度、鸡只健康以及产蛋情况。
表1 基础饲粮组成及营养水平(饲喂基础)

Table 1 Composition and nutrient levels of the basal diet (as-fed basis) %

项目Items 含量Content
原料Ingredients
玉米Corn 59.40
豆粕Soybean meal 26.50
大豆油Soybean oil 3.00
磷酸氢钙CaHPO4 2.35
石粉Limestone 2.05
贝壳粉Shell powder 5.10
氯化钠NaCl 0.30
DL-蛋氨酸DL-Met 0.20
预混料Premix1) 1.00
玉米淀粉或甜菜碱Cornstarch or betaine2) 0.10
合计Total 100.00
营养水平Nutrient levels3)
代谢能ME/(MJ/kg) 11.51
粗蛋白质CP 16.57
赖氨酸Lys 0.82
蛋氨酸Met 0.42
蛋氨酸+胱氨酸Met+Cys 0.66
钙Ca 3.20
非植酸磷NPP 0.38

1)预混料为每千克饲粮提供 Premix provided the following per kilogram of the diet:VA 7 715 IU,VD3 5 000 IU,VE 10 IU,VK3 2.2 mg,VB1 1.3 mg,VB2 3.4 mg,VB6 3.25 mg,VB12 0.02 mg,泛酸钙 calcium pantothenate 10 mg,烟酸 niacin 33 mg,叶酸 folic acid 0.28 mg,生物素 biotin 0.2 mg,胆碱 choline 600 mg,Cu (CuSO4·5H2O) 8 mg,Mn (MnSO4·H2O) 80 mg,Zn (ZnSO4·H2O) 80 mg,Se (Na2SeO3) 0.3 mg,I [Ca(IO3)2·H2O] 0.8 mg。

2)甜菜碱+皮质酮组按替代等重的玉米淀粉添加甜菜碱。Betaine supplement added in place of equivalent weights of cornstarch in betaine+corticosterone group.

3)粗蛋白质和钙为实测值,其他营养水平为计算值。CP and Ca were measured values, while the other nutrient levels were calculated values.

1.4 样品采集与制备

试验开始前,将购买的基础饲粮现场采样,研磨过200目筛后置于自封袋中,于-20 ℃保存,用于分析饲粮中粗蛋白质和钙含量。
试验至35 d时,所有试验鸡只禁食不禁水12 h后,每个重复内选取2只试验鸡,然后宰杀,分离其十二指肠、回肠和空肠肠段,将肠道内容物用冰生理盐水冲洗干净,使用已灭菌的载玻片刮取肠黏膜,立即将其置于灭菌的1.5 mL离心管中,于-196 ℃液氮速冻后保存于-80 ℃冰箱,以备检测炎症因子IL-1β和白细胞介素-6(interleukin-6,IL-6)含量及Toll样受体1(Toll-like receptor 1,TLR1)、Toll样受体2(Toll-like receptor 2,TLR2)、Toll样受体4(Toll-like receptor 4,TLR4)、IL-1β、IL-6、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-10(interleukin-10,IL-10)、白细胞介素-8(Interleukin-8,IL-8)和核因子-κB p65(nuclear factor-kappa B p65,NF-κB p65)的mRNA表达水平。最后,将2只试验鸡的组织样品按等比例合并为1个样本进行分析。

1.5 指标测定

用凯氏定氮法(GB/T 6432—1994)[10]测定饲粮中粗蛋白质含量,采用原子吸收光谱法(GB/T 13885—2017)[11]测定饲粮中钙含量。饲粮其他营养水平参照《中国饲料成分及营养价值表》(2022年第33版)计算。
采用酶联免疫吸附测定试剂盒(北京索莱宝生物科技有限公司)检测十二指肠、空肠和回肠中炎症因子IL-1β(SEKN-0153)和IL-6(SEKCN-0161)含量,检测方法均按照试剂盒说明书步骤进行。
采用SYBR Green RT-qPCR法测定十二指肠、空肠和回肠黏膜中TLR1、TLR2、TLR4、TNF-α、IL-6、IL-8、IL-10、IL-1β和NF-κB p65的mRNA表达水平。使用双内参法,2个内参基因分别为甘油醛-3-磷酸脱氢酶(GAPDH)和β-肌动蛋白(β-actin);采用2-ΔΔCt法计算各个目的基因的mRNA表达水平,引物序列信息见表2。
表2 引物序列信息

Table 2 Primer sequence information

基因
Genes		 GenBank登录号
GenBank accession number		 引物序列
Primer sequences (5’—3’)		 产物长度
Product length/bp
白细胞介素-1β
IL-1β		 NM_204524.2 F:CAAACTGCTGCGGAGGC
R:AGGACTGTGAGCGGGTGTAG		 149
肿瘤坏死因子-α
TNF-α		 NM_204267.2 F:GCCCTTCCTGTAACCAGATG
R:ACACGACAGCCAAGTCAACG		 208
白细胞介素-6
IL-6		 NM_204628.2 F:AGGACGAGATGTGCAAGAAGT
R:TTGGGCAGGTTGAGGTTGTT		 130
白细胞介素-8
IL-8		 NM_205018.2 F:CAAGCCAAACACTCCT
R:TTTAGGGTGGATGAACT		 158
白细胞介素-10
IL-10		 NM_001004414.4 F:GCTGAGGGTGAAGTTTGAG
R:CAGGTGAAGAAGCGGTGA		 278
Toll样受体1
TLR1		 NM_001007488.5 F:ACCCGTTCAAGTGTTCGTG
R:TTCCGCTCAAGTCTTCTGG		 120
Toll样受体2
TLR2		 NM_001161650.3 F:GGTGTTCCTGTTCATCCTCATC
R:GTTGGAGTCGTTCTCACTGTAGG		 169
Toll样受体4
TLR4		 NM_001030693.2 F:ATCTTTCAAGGTGCCACATC
R:GGATATGCTTGTTTCCACCA		 215
核因子-κB p65
NF-κB p65		 NM_001396038.1 F:AGGACTTAAAATGGCAGGAGA
R:GCTGTTCGTAGTGGTAAGTCT		 225

1.6 数据统计分析

采用SAS 9.4统计软件中一般线性模型(GLM)对试验数据进行单因素方差分析。方差分析显著者,以最小显著差异(LSD)法比较各平均值间的差异显著性。鸡只死亡率数据经反正弦转换后进行统计分析。以重复作为1个试验单元。结果以平均值和集合标准误表示,P<0.05作为差异显著性检验标准。

2 结果与分析

2.1 甜菜碱对皮质酮诱导的产蛋鸡体重、产蛋性能及死亡率的影响

课题组前期研究发现,皮质酮和甜菜碱处理对产蛋鸡的体重、平均日采食量、蛋重和死亡率没有显著影响(P>0.05),但对产蛋率和料蛋比有显著影响(P<0.05)。与对照组相比,皮质酮组的产蛋率显著降低(P<0.05),料蛋比显著提高(P<0.05);与皮质酮组相比,饲粮添加甜菜碱对皮质酮诱导的产蛋性能降低未表现出显著改善作用(P>0.05)[12]。

2.2 甜菜碱对皮质酮诱导的产蛋鸡十二指肠、空肠和回肠中IL-1β和IL-6含量的影响

由表3可知,皮质酮和甜菜碱处理对产蛋鸡十二指肠、空肠和回肠中IL-1β含量有显著影响(P<0.05),但对IL-6含量没有显著影响(P>0.05)。与对照组相比,皮质酮组的十二指肠、空肠和回肠中IL-1β含量显著升高(P<0.05);与皮质酮组相比,甜菜碱+皮质酮组的十二指肠、空肠和回肠中IL-1β含量显著降低(P<0.05),说明饲粮添加甜菜碱缓解了皮质酮诱导的产蛋鸡十二指肠、空肠和回肠中IL-1β含量的升高。
表3 甜菜碱对皮质酮诱导的产蛋鸡十二指肠、空肠和回肠中IL-1β和IL-6含量的影响

Table 3 Effects of betaine on contents of IL-1β and IL-10 in duodenum, jejunum and ileum of laying hens induced by corticosterone pg/mg prot

项目
Items		 十二指肠Duodenum 空肠Jejunum 回肠Ileum
白细胞
介素-1β
IL-1β		 白细胞
介素-6
IL-6		 白细胞
介素-1β
IL-1β		 白细胞
介素-6
IL-6		 白细胞
介素-1β
IL-1β		 白细胞
介素-6
IL-6
对照组CON group 279c 25.2 281b 22.5 402b 32.7
皮质酮组CORT group 436a 31.4 397a 23.5 520a 47.5
甜菜碱+皮质酮组BET+CORT group 365b 29.8 332b 26.7 404b 34.4
集合标准误Pooled SE 13 2.9 28 3.2 34 5.6
P值P-value 0.000 1 0.342 3 0.007 9 0.633 4 0.039 2 0.154 1

同列数据肩标不同小写字母表示差异显著(P<0.05),相同或无字母表示差异不显著(P>0.05)。

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

2.3 甜菜碱对皮质酮诱导的产蛋鸡十二指肠中炎症相关基因mRNA表达水平的影响

由表4可知,皮质酮和甜菜碱处理对产蛋鸡十二指肠中IL-8、NF-κB p65和TLR4的mRNA表达水平有显著影响(P<0.05),但对TLR1、TLR2、TNF-α、IL-1β、IL-6和IL-10的mRNA表达水平没有显著影响(P>0.05)。与对照组相比,皮质酮组的十二指肠中TLR4、IL-8和NF-κB p65的mRNA表达水平显著升高(P<0.05);与皮质酮组相比,甜菜碱+皮质酮组的十二指肠中TLR4、IL-8和NF-κB p65的mRNA表达水平显著降低(P<0.05),说明饲粮添加甜菜碱缓解了皮质酮诱导的产蛋鸡十二指肠中TLR4、IL-8和NF-κB p65的mRNA表达水平的升高。
表4 甜菜碱对皮质酮诱导的产蛋鸡十二指肠中炎症相关基因mRNA表达水平的影响

Table 4 Effects of betaine on mRNA expression levels of inflammation related genes in duodenum of laying hens induced by corticosterone

项目
Items		 对照组
CON group		 皮质酮组
CORT group		 甜菜碱+皮质酮组
BET+CORT group		 集合标准误
Pooled SE		 P值
P-value
Toll样受体1 TLR1 1.01 1.19 1.02 0.04 0.173 0
Toll样受体2 TLR2 1.01 1.12 1.08 0.04 0.541 7
Toll样受体4 TLR4 1.02b 1.26a 0.94b 0.07 0.018 3
肿瘤坏死因子-α TNF-α 1.02 1.03 1.02 0.07 0.997 5
白细胞介素-1β IL-1β 1.01 1.13 1.01 1.01 0.147 8
白细胞介素-6 IL-6 1.06 1.26 1.17 0.08 0.227 0
白细胞介素-8 IL-8 1.02b 1.76a 0.98b 0.07 0.000 1
白细胞介素-10 IL-10 1.01 0.97 1.00 0.04 0.804 1
核因子-κB p65 NF-κB p65 1.01b 1.33a 1.01b 0.03 0.000 1

同行数据肩标不同小写字母表示差异显著(P<0.05),相同或无字母表示差异不显著(P>0.05)。下表同。

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

2.4 甜菜碱对皮质酮诱导的产蛋鸡空肠中炎症相关基因mRNA表达水平的影响

由表5可知,皮质酮和甜菜碱处理对产蛋鸡空肠中IL-1β、IL-10和NF-κB p65的mRNA表达水平有显著影响(P<0.05),但对TLR1、TLR2、TLR4、TNF-α、IL-6和IL-8的mRNA表达水平没有显著影响(P>0.05)。与对照组相比,皮质酮组的空肠中IL-1β和NF-κB p65的mRNA表达水平显著升高(P<0.05);与皮质酮组相比,甜菜碱+皮质酮组的空肠中IL-1β的mRNA表达水平显著降低(P<0.05),IL-10的mRNA表达水平显著升高(P<0.05),说明饲粮添加甜菜碱缓解了皮质酮诱导的产蛋鸡空肠中IL-1β的mRNA表达水平的升高,且进一步提高了IL-10的mRNA表达水平。
表5 甜菜碱对皮质酮诱导的产蛋鸡空肠中炎症相关基因mRNA表达水平的影响

Table 5 Effects of betaine on mRNA expression levels of inflammation related genes in jejunum of laying hens induced by corticosterone

项目
Items		 对照组
CON group		 皮质酮组
CORT group		 甜菜碱+皮质酮组
BET+CORT group		 集合标准误
Pooled SE		 P值
P-value
Toll样受体1 TLR1 1.03 0.98 0.86 0.06 0.537 5
Toll样受体2 TLR2 1.03 0.89 0.86 0.06 0.468 1
Toll样受体4 TLR4 1.00 1.11 0.92 0.07 0.272 5
肿瘤坏死因子-α TNF-α 1.03 1.09 0.97 0.07 0.538 7
白细胞介素-1β IL-1β 1.03b 1.52a 1.23b 0.07 0.007 9
白细胞介素-6 IL-6 1.03 1.27 0.93 0.09 0.055 4
白细胞介素-8 IL-8 1.01 1.10 1.11 0.04 0.561 8
白细胞介素-10 IL-10 1.02b 1.25b 1.67a 0.08 0.000 7
核因子-κB p65 NF-κB p65 1.01b 1.59a 1.47a 0.14 0.023 4

2.5 甜菜碱对皮质酮诱导的产蛋鸡回肠中炎症相关基因mRNA表达水平的影响

由表6可知,皮质酮和甜菜碱处理对产蛋鸡回肠中IL-1β、IL-8和NF-κB p65的mRNA表达水平有显著影响(P<0.05),但对TLR1、TLR2、TLR4、TNF-α、IL-6和IL-10的mRNA表达水平没有显著影响(P>0.05)。与对照组相比,皮质酮组的回肠中IL-1β、IL-8和NF-κB p65的mRNA表达水平显著升高(P<0.05);与皮质酮组相比,甜菜碱+皮质酮组的空肠中IL-1β、IL-8和NF-κB p65 mRNA表达水平显著降低(P<0.05),说明饲粮添加甜菜碱缓解了皮质酮诱导的产蛋鸡空肠中IL-1β、IL-8和NF-κB p65的mRNA表达水平的升高。
表6 甜菜碱对皮质酮诱导的产蛋鸡回肠中炎症相关基因mRNA表达水平的影响

Table 6 Effects of betaine on mRNA expression levels of inflammation related genes in ileum of laying hens induced by corticosterone

项目
Items		 对照组
CON group		 皮质酮组
CORT group		 甜菜碱+皮质酮组
BET+CORT group		 集合标准误
Pooled SE		 P值
P-value
Toll样受体1 TLR1 1.06 1.01 1.03 0.06 0.898 0
Toll样受体2 TLR2 1.01 1.00 0.93 0.07 0.797 2
Toll样受体4 TLR4 0.98 1.11 0.92 0.07 0.272 5
肿瘤坏死因子-α TNF-α 0.99 1.02 0.98 0.05 0.863 8
白细胞介素-1β IL-1β 0.90b 1.70a 1.16b 0.08 0.000 3
白细胞介素-6 IL-6 1.05 1.31 1.28 0.16 0.508 1
白细胞介素-8 IL-8 1.02b 1.46a 1.13b 0.06 0.000 2
白细胞介素-10 IL-10 1.01 0.85 0.93 0.04 0.078 8
核因子-κB p65 NF-κB p65 1.01b 1.21a 0.85c 0.05 0.000 2

3 讨论

先前研究表明,甜菜碱可有效缓解应激或高脂诱导的动物肝脏炎症反应[5,13]。本试验研究发现,饲粮添加甜菜碱可通过降低产蛋鸡小肠各段中IL-1β含量及炎症相关基因mRNA表达水平,进而改善皮质酮诱导的产蛋鸡小肠炎症反应。该研究为甜菜碱在生产中的合理应用和缓解应激诱导的产蛋鸡小肠炎症反应提供了科学试验依据。
研究表明,动物受到病毒感染或遭受慢性应激时会激活巨噬细胞释放IL-1β和IL-6等炎症因子,引发炎症反应[14]。因此,血清或组织中IL-1β等炎症因子的含量可作为判定动物机体是否发生炎症反应的重要参考指标。Feng等[15]研究发现,慢性皮质酮处理显著促进了鸡肝脏炎症反应,伴随着血清中IL-1β含量显著升高,表明长期的慢性应激可诱导动物机体发生炎症反应。Alhotan等[16]研究发现,热应激显著提高了肉仔鸡空肠中IL-1β含量,进而诱发了空肠的炎症反应;而饲粮添加甜菜碱可通过缓解热应激诱导的肉鸡空肠中IL-1β含量的升高来减缓空肠的炎症反应。与前人研究结果相似,本研究发现,皮质酮处理显著提高了如皋黄鸡十二指肠、空肠和回肠中炎症因子IL-1β含量,而甜菜碱可有效缓解皮质酮诱导的产蛋鸡十二指肠、空肠和回肠中IL-1β含量的升高,表明饲粮添加甜菜碱可有效缓解皮质酮诱导的产蛋鸡小肠炎症反应。
Toll样受体是一类模式识别受体,位于细胞膜上,可识别不同的外界刺激,并招募特异性蛋白,进而激活一系列信号级联反应,调控组织的炎性反应[17-18]。NF-κB p65是Toll样受体的下游蛋白分子,当动物发生炎症反应或受到氧化应激时,体内的炎症因子会激活NF-κB信号通路,使NF-κB p65转移到细胞核中,提高动物体内TNF-α、IL-1β和IL-6等炎症因子的转录表达水平,进一步加剧动物炎症反应[19]。Feng等[15]研究发现,慢性皮质酮处理显著促进鸡肝脏中炎症相关基因TLR1、TLR4、NF-κB p65及IL-1β等的mRNA表达水平,进而促进了鸡肝脏的炎症反应。李鼎等[5]研究表明,注射皮质酮显著提高了产蛋鸡肝脏中IL-1β的mRNA表达水平,造成肝脏炎症损伤。与前人研究结果相似,本研究发现,皮质酮处理显著提高了产蛋鸡十二指肠中TLR4、IL-8和NF-κB p65,空肠中IL-1β和NF-κB p65以及回肠中IL-1β、IL-8和NF-κB p65的mRNA表达水平,提示慢性应激可通过促进炎症相关基因的表达来加剧小肠的炎症反应。因此,抑制小肠炎症相关基因的表达对于缓解应激诱导的小肠炎症反应具有重要的调控作用。然而,在本研究中,不同小肠段中炎症相关因子的mRNA表达水平变化并不一致,可能与不同小肠段对应激反应的敏感性不同有关。
在哺乳动物和鸡上的研究均发现,甜菜碱可以通过抑制炎症相关基因的表达来缓解动物机体的炎症反应[5,20]。Zhang等[21]研究发现,甜菜碱可有效缓解β淀粉样蛋白1-42寡聚体诱导的小胶质细胞中IL-1β、IL-8、核因子-κB(NF-κB)和TNF-α的mRNA表达水平的升高,进而缓解了β淀粉样蛋白1-42寡聚体诱导的小胶质细胞炎症反应。阳金金等[22]研究表明,饲粮添加甜菜碱可通过降低脾脏中IL-1β的mRNA表达水平缓解LPS诱导的鹅脾脏炎症损伤。Zhang等[23]研究表明,甜菜碱可通过降低雏鸡法氏囊中IL-6的mRNA表达水平缓解LPS诱导的法氏囊炎症反应。本研究结果表明,甜菜碱可显著缓解皮质酮诱导的产蛋鸡十二指肠中TLR4、IL-8和NF-κB p65,空肠中IL-1β和NF-κB p65以及回肠中IL-1β、IL-8和NF-κB p65的mRNA表达水平的升高,进而缓解了皮质酮诱导的小肠炎症反应。此外,本研究还发现,与皮质酮组相比,皮质酮+甜菜碱组的空肠中IL-10的mRNA表达水平显著提高。IL-10是一种抗炎细胞因子,能有效地抑制促炎细胞因子的活性,改善机体的炎症反应。上述研究结果表明,甜菜碱可通过降低小肠中促炎相关基因的表达和提高抗炎相关基因的表达,进而缓解皮质酮诱导的产蛋鸡小肠炎症反应。

4 结论

饲粮中添加0.1%甜菜碱通过降低产蛋鸡十二指肠、空肠和回肠中IL-1β含量及促炎相关基因的mRNA表达水平,缓解皮质酮诱导的产蛋鸡小肠炎症反应。

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