上笼应激对绍兴鸭十二指肠组织结构、抗氧化能力及基因mRNA表达量的影响

doi:10.3785/j.issn.1008-9209.2019.06.181

浙江大学学报（农业与生命科学版）

2020, Vol. 46

Issue (2): 234-242 DOI: 10.3785/j.issn.1008-9209.2019.06.181

动物科学与动物医学

上笼应激对绍兴鸭十二指肠组织结构、抗氧化能力及基因mRNA表达量的影响

顾天天^1,²(

),田勇¹,周玮³,刘国发³,陈黎¹,曾涛¹,吴信生²,徐琪²,陈国宏²(

),卢立志¹(

)

^1.浙江省农业科学院畜牧兽医研究所，杭州 310021
^2.扬州大学动物科学与技术学院，江苏扬州 225009
^3.周口桂柳种鸭育种有限公司，河南周口 461300

Effects of caged stress on the duodenal tissue structure, antioxidant capacity and gene mRNA expression level of Shaoxing duck

Tiantian GU^1,²(

),Yong TIAN¹,Wei ZHOU³,Guofa LIU³,Li CHEN¹,Tao ZENG¹,Xinsheng WU²,Qi XU²,Guohong CHEN²(

),Lizhi LU¹(

)

^1.Institute of Animal Husbandry and Veterinary Science, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
^2.College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, Jiangsu, China
^3.Zhoukou Guiliu Duck Breeding Co. , Ltd. , Zhoukou 461300, Henan, China

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摘要：

将80只200日龄的绍兴蛋鸭随机分为2组，即地面平养组和上笼应激组，在上笼后第1、2、4、7和10天对十二指肠的组织损伤程度以及抗氧化等指标进行测定，并通过实时荧光聚合酶链式反应（real-time fluorescence quantitative polymerase chain reaction, RT-PCR）检测内质网应激、炎症和凋亡相关基因mRNA的相对表达量，为蛋鸭的规模化养殖和上笼应激诱发症的科学防预提供理论依据。结果发现：1）随着上笼应激时间的增加，试验组十二指肠的损伤程度逐渐增加，主要表现为炎症细胞浸润和肠腺上皮细胞脱落。 2）上笼应激过程中，试验组十二指肠丙二醛（malondialdehyde, MDA）含量,总抗氧能力（total antioxidant capacity, T-AOC）、超氧化物歧化酶（superoxide dismutase, SOD）和谷胱甘肽过氧化物酶（glutathione peroxidase, GSH-Px）活性呈现波动性变化，而过氧化氢酶（catalase, CAT）活性在上笼应激后第7天显著增加（P＜0.05）。3）RT-PCR检测结果显示：与对照组相比，内质网应激相关基因葡萄糖调节蛋白78（glucose regulated protein 78, GRP78）和CCAAT/增强子结合蛋白同源蛋白（CCAAT/enhancer-binding protein-homologous protein, CHOP） mRNA表达水平呈现波动性变化，且在上笼后第2和10天显著增加（P＜0.05）;凋亡相关基因Bcl-2相关X蛋白（Bcl-2 associated X protein, Bax） mRNA表达水平在上笼后第7和10天显著增加（P＜0.05），而半胱氨酸天冬氨酸特异性蛋白酶3（cysteinecontaining aspartate-specific proteases-3, Caspase3） mRNA表达量较对照组差异不显著（P＞0.05）；炎症相关基因环氧合酶-2（cyclooxygenase-2, COX-2） mRNA水平显著上调，且在上笼后第10天显著增加（P＜0.05），而诱导型一氧化氮合酶（inducible nitric oxide, iNOS）表达水平则呈现先下降后上升的趋势。综上表明，上笼应激过程引起绍兴鸭十二指肠不同程度的组织损伤，同时内质网应激强度上调，最终导致细胞凋亡和炎症反应的产生。

关键词： 上笼应激; 绍兴鸭; 十二指肠; 抗氧化能力; 基因表达

Abstract:

Eighty 200-day-old Shaoxing ducks were randomly divided into two groups, including the ground group (control group) and the caged stress group (treatment group). Then the tissue damage degree of the duodenum and antioxidant indicators were determinated, and the relative expression levels of endoplasmic reticulum stress gene, inflammation-related gene and apoptotic gene mRNA were detected by real-time fluorescence polymerase chain reaction (RT-PCR) at 1, 2, 4, 7 and 10 d after cage rearing, which providing theoretical basis for the large-scale breeding of laying ducks and the scientific prevention for caged stress disease. The results showed that: 1) With the increase of caged stress time, the damage degree of the duodenum in the treatment group gradually increased, mainly manifested by inflammatory cell infiltration and intestinal gland epithelial cell shedding. 2) During the progress of caged stress, the duodenal malondialdehyde (MDA) content, total antioxidant capacity (T-AOC), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), activities in the treatment group showed fluctuating changes, while the catalase (CAT) activity increased significantly at the 7th day after cage rearing (P＜0.05). 3) RT-PCR results showed that compared with the control group, the endoplasmic reticulum stress-related genes glucose regulated protein 78 (GRP78)and CCAAT/enhancer-binding protein-homologous protein (CHOP) mRNA expression levels showed fluctuating changes, and increased significantly in the Shaoxing ducks at the 2nd and 10th day after cage rearing (P＜0.05). In addition, the expression level of apoptosis-related gene Bcl-2 associated X protein (Bax) mRNA increased significantly at the 7th and 10th day after cage rearing (P＜0.05), while the expression of cysteinecontaining aspartate-specific proteases-3 (Caspase3) mRNA expression level was not significantly different from the control group (P＞0.05). At the same time, the cyclooxygenase-2 (COX-2) mRNA expression level was significantly up-regulated and increased significantly at the 10th day after cage rearing (P＜0.05), while the expression level of inducible nitric oxide (iNOS) showed a trend of decreasing first and then increasing. The above results indicate that the stress of caged rearing causes different degrees of tissue damage on Shaoxing duck duodenum, which enhances the body’s antioxidant capacity. At the same time, the up-regulation of endoplasmic reticulum stress intensity eventually leads to apoptosis and inflammation.

Key words: caged stress Shaoxing duck duodenum antioxidant capacity gene expression

收稿日期: 2019-06-18 出版日期: 2020-05-22

CLC:

S 834

基金资助: 国家重点研发计划(2018YFD0501504);国家自然科学基金(31702106);浙江省自然科学基金(LQ17C170003);国家水禽产业技术体系专项(CARS-43-6);浙江省基础公益研究计划(LGN18C170003)

通讯作者: 陈国宏,卢立志 E-mail: gtt19931029@126.com;ghchen@yzu.edu.cn;lulizhibox@ 163.com

作者简介: 顾天天（https://orcid.org/0000-0002-4430-9320），E-mail：gtt19931029@126.com

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引用本文:

顾天天,田勇,周玮,刘国发,陈黎,曾涛,吴信生,徐琪,陈国宏,卢立志. 上笼应激对绍兴鸭十二指肠组织结构、抗氧化能力及基因mRNA表达量的影响[J]. 浙江大学学报（农业与生命科学版）, 2020, 46(2): 234-242.

Tiantian GU,Yong TIAN,Wei ZHOU,Guofa LIU,Li CHEN,Tao ZENG,Xinsheng WU,Qi XU,Guohong CHEN,Lizhi LU. Effects of caged stress on the duodenal tissue structure, antioxidant capacity and gene mRNA expression level of Shaoxing duck. Journal of Zhejiang University (Agriculture and Life Sciences), 2020, 46(2): 234-242.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2019.06.181 或 http://www.zjujournals.com/agr/CN/Y2020/V46/I2/234

表1 RT-PCR引物信息

图1 上笼应激对绍兴鸭十二指肠组织结构的影响A～E.对照组；F～J.处理组。黑色箭头表示脱落的肠腺上皮细胞。

图2 上笼应激对绍兴鸭十二指肠抗氧化能力的影响

图3 上笼应激对绍兴鸭十二指肠应激相关基因mRNA表达量的影响*表示同一应激时间不同处理间在P＜0.05水平差异有统计学意义。

图4 上笼应激对绍兴鸭十二指肠凋亡相关基因mRNA表达量的影响*表示同一应激时间不同处理间在P＜0.05水平差异有统计学意义。

图5 上笼应激对绍兴鸭十二指肠炎症相关基因mRNA表达量的影响*表示同一应激时间不同处理间在P＜0.05水平差异有统计学意义。

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