猪流行性腹泻病毒感染对仔猪肠道损伤及肠道细胞巨泡式死亡的影响

doi:10.11843/j.issn.0366-6964.2025.12.028

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (12): 6271-6280.doi: 10.11843/j.issn.0366-6964.2025.12.028

猪流行性腹泻病毒感染对仔猪肠道损伤及肠道细胞巨泡式死亡的影响

陈琳琳¹^,²(), 著龙祥¹^,²(), 祖少坡¹^,², 陈红英¹^,², 李庆豪¹^,², 金鑫¹^,², 李艺雷¹^,²^,*(), 马世杰¹^,²^,*(), 魏战勇¹^,²

1. 河南农业大学动物医学院, 郑州 450046
2. 动物病原与生物安全教育部重点实验室, 郑州 450046

收稿日期:2025-03-05 出版日期:2025-12-23 发布日期:2025-12-24
通讯作者: 李艺雷,马世杰 E-mail:ch748250@126.com;1197842170@qq.com;liyilei@henau.edu.cn;participate2006@126.com
作者简介:陈琳琳(2000-)，女，河南许昌人，硕士，主要从事猪冠状病毒致病机制研究，E-mail：ch748250@126.com
著龙祥(2001-)，河南平顶山人，硕士，主要从事猪冠状病毒致病机制研究，E-mail：1197842170@qq.com
第一联系人：
陈琳琳和著龙祥为同等贡献作者
基金资助:
国家自然科学基金重点项目(32130106)；国家自然科学基金区域创新发展联合基金重点项目(U22A20520)；河南省自然科学基金面上项目(242300421324)；河南农业大学科技创新基金项目(30501649)

Effects of Porcine Epidemic Diarrhea Virus Infection on Intestinal Injury and Intestinal Cell Methuosis in Piglet

CHEN Linlin¹^,²(), ZHU Longxiang¹^,²(), ZU Shaopo¹^,², CHEN Hongying¹^,², LI Qinghao¹^,², JIN Xin¹^,², LI Yilei¹^,²^,*(), MA Shijie¹^,²^,*(), WEI Zhanyong¹^,²

1. College of Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China
2. Ministry of Education Key Laboratory for Animal Pathogens and Biosafety, Zhengzhou 450046, China

Received:2025-03-05 Online:2025-12-23 Published:2025-12-24
Contact: LI Yilei, MA Shijie E-mail:ch748250@126.com;1197842170@qq.com;liyilei@henau.edu.cn;participate2006@126.com

摘要/Abstract

摘要：

为了探讨猪流行性腹泻病毒(porcine epidemic diarrhea virus，PEDV)新型野毒株对初生仔猪肠道损伤及肠道细胞巨泡式死亡的影响，选取8头5日龄的健康仔猪，随机分为空白对照组(Mock组)和PEDV感染组，病毒感染后记录临床症状并在2 d后实施安乐死，之后分别采集十二指肠、空肠、回肠、盲肠、结肠和直肠的组织样品。采用苏木精-伊红(HE)染色、免疫组织化学(IHC)染色、RT-qPCR和Western blot技术检测PEDV感染仔猪后各肠段的组织病理学变化、病毒在肠道中的定位及对巨泡式死亡的影响。HE染色结果显示，PEDV对仔猪各肠段黏膜结构均有不同程度的损伤，其中对空肠的损伤最为严重。RT-qPCR结果证实PEDV感染仔猪后，空肠中病毒载量最高。IHC染色发现PEDV主要分布在小肠黏膜的上皮层中，其中在空肠中阳性信号最多，与RT-qPCR结果一致。深入研究揭示PEDV感染组的空肠组织中巨泡式死亡相关基因Rac1(Ras相关C3肉毒菌毒素底物1)、Rab7(Ras相关蛋白7)和LAMP1(溶酶体相关膜蛋白1)的表达量显著上调，Western blot试验也进一步证实了这一发现。以上结果表明，PEDV新型野毒株感染仔猪后主要定位于小肠，对空肠的损伤最严重，并使空肠发生巨泡式死亡。本研究可为深入理解PEDV的感染发病机制提供可靠的参考依据。

关键词: 猪流行性腹泻病毒, 空肠, 病理损伤, 巨泡式死亡

Abstract:

To investigate the effects of new wild-type PEDV infection on the intestinal mucosal injury and intestinal cell methuosis of piglets, 8 healthy piglets aged 5 days were randomly divided into Mock and PEDV-infected groups. The clinical symptoms were monitored. After 2 days of virus infection, the piglets were euthanized and the duodenum, jejunum, ileum, cecum, colon, and rectum were collected. Hematoxylin-eosin (HE) staining, immunohistochemistry (IHC) staining, RT-qPCR and Western blot were used to detect the pathological changes of the intestinal tissues, the viral location in the intestines, and its effect on methuosis. HE staining revealed that PEDV-infected group had different degree of damage to the intestinal mucosa, and the jejunum showed more obvious pathological damage. Compared with other intestinal segments, the viral load was the highest in jejunum after PEDV infection using RT-qPCR. IHC staining showed that PEDV mainly distributed in the upper cortex of small intestinal mucosa, and the most positive signals were observed in jejunum segment, which was consistent with the result of RT-qPCR. Further study revealed that the expression levels of megavesicular death-related genes Rac1(Ras-related C3 sarcoma virus oncogene 1), Rab7(Ras-related protein 7) and LAMP1(lysosomal-related membrane protein 1) in PEDV infection group increased significantly in jejunum, which was further confirmed by Western blot. The above results indicate that the new wild-type PEDV mainly located in the small intestine, caused the most serious pathological damage to jejunum and induced methuosis of the jejunal cells. This study could provide a reliable basis for further understanding of the pathogenesis of PEDV infection.

Key words: porcine epidemic diarrhea virus, jejunum, pathological injury, methuosis

中图分类号:

S852.659.6

陈琳琳, 著龙祥, 祖少坡, 陈红英, 李庆豪, 金鑫, 李艺雷, 马世杰, 魏战勇. 猪流行性腹泻病毒感染对仔猪肠道损伤及肠道细胞巨泡式死亡的影响[J]. 畜牧兽医学报, 2025, 56(12): 6271-6280.

CHEN Linlin, ZHU Longxiang, ZU Shaopo, CHEN Hongying, LI Qinghao, JIN Xin, LI Yilei, MA Shijie, WEI Zhanyong. Effects of Porcine Epidemic Diarrhea Virus Infection on Intestinal Injury and Intestinal Cell Methuosis in Piglet[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(12): 6271-6280.

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引物名称 Primer name	引物序列(5′→3′) Primer sequence	产物大小/bp Product size
PEDV-M-F	CTCGGCTTGCATCACTCT	216
PEDV-M-R	GACCCAGTAGCAACCTTAT
β-actin-F	CCCTGGAGAAGAGCTACGAG	175
β-actin-R	AGGTCCTTCCTGATGTCCAC
Rac1-F	AGGGATGATAAAGACACGAT	111
Rac1-R	CTCCAGGTATTTCACAGCA
Rab7-F	CAAAGCGACAATAGGAGC	151
Rab7-R	CACCAGAACGCAGCAGT
LAMP1-F	GGGACCGCCTGCATCAT	215
LAMP1-R	CGGGTGGCATTTCTTGTG

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猪流行性腹泻病毒感染对仔猪肠道损伤及肠道细胞巨泡式死亡的影响

Effects of Porcine Epidemic Diarrhea Virus Infection on Intestinal Injury and Intestinal Cell Methuosis in Piglet

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