ACE2对猪流行性腹泻病毒体外感染传代猪小肠上皮细胞的影响

doi:10.11843/j.issn.0366-6964.2024.03.035

畜牧兽医学报 ›› 2024, Vol. 55 ›› Issue (3): 1238-1248.doi: 10.11843/j.issn.0366-6964.2024.03.035

ACE2对猪流行性腹泻病毒体外感染传代猪小肠上皮细胞的影响

任莉鑫¹, 张静怡¹, 徐沙沙¹, 杨柳², 张兴翠^1*, 宋振辉^1,3*

1. 西南大学动物医学院, 重庆 402460;
2. 重庆市畜牧科学院, 重庆 402460;
3. 西南大学医学研究院免疫学研究中心, 重庆 402460

收稿日期:2023-04-13 出版日期:2024-03-23 发布日期:2024-03-27
通讯作者: 张兴翠,主要从事动物微生物学与免疫学研究,E-mail:zhangxc923@163.com;宋振辉,主要从事动物微生物学与免疫学研究,E-mail:szh7678@126.com
作者简介:任莉鑫(1999-),女,河北秦皇岛人,硕士生,主要从事动物病原微生物与免疫研究,E-mail:525104823@qq.com;张静怡(1998-),女,山东淄博人,硕士,主要从事动物病原微生物与免疫研究,E-mail:564461985@qq.com。
基金资助:
中央高校基本业务基金资助项目（XDJK2020RC001）；重庆市研究生科研创新项目（CYS19137）；国家生猪技术创新中心项目（NCTIP-XD/Bn）

Effect of ACE2 on Porcine Intestinal Epithelial Cells Infected with Porcine Epidemic Diarrhea Virus in vitro

REN Lixin¹, ZHANG Jingyi¹, XU Shasha¹, YANG Liu², ZHANG Xingcui^1*, SONG Zhenhui^1,3*

1. College of Veterinary Medicine, Southwest University, Chongqing 402460, China;
2. Chongqing Academy of Animal Husbandry, Chongqing 402460, China;
3. Immunology Research Center, Institute of Medical Research, Southwest University, Chongqing 402460, China

Received:2023-04-13 Online:2024-03-23 Published:2024-03-27

摘要/Abstract

摘要： 为探究血管紧张素转换酶2（angiotensin converting enzyme 2，ACE2）在猪流行性腹泻病毒（porcine epidemic diarrhea virus，PEDV）感染仔猪过程中发挥的作用，本研究通过转录组学分析仔猪肠道ACE2在PEDV感染前后表达的变化情况，然后利用猪小肠上皮细胞（porcine intestinal epithelial cells，IPEC-J2细胞）模型，通过RT-qPCR、Western blot检测PEDV感染后ACE2的mRNA、蛋白表达水平的变化。过表达与抑制表达ACE2后通过RT-qPCR、Western blot、TCID₅₀检测PEDV复制水平。结果显示，PEDV感染IPEC-J2后ACE2的mRNA及蛋白表达水平均显著下调，与转录组学结果一致。过表达ACE2组PEDV感染量显著上升，抑制表达ACE2组PEDV感染量显著下降。本研究在细胞水平上验证了ACE2在PEDV感染过程中的作用，即PEDV感染能够使ACE2表达量下降，在IPEC-J2细胞中过表达ACE2能提高PEDV复制水平，抑制ACE2的表达可降低PEDV复制水平。

关键词: 血管紧张素转换酶2, ACE2, 猪流行性腹泻病毒, 猪小肠上皮细胞

Abstract: The present study aimed to investigate the role of angiotensin converting enzyme 2 (ACE2) in the infection of piglets with porcine epidemic diarrhea virus (PEDV). We investigated the expression of ACE2 before and after PEDV infection in piglets by transcriptomic analysis. Then the changes of ACE2 expression before and after PEDV infection were verified by using Porcine Intestinal Epithelial Cells (IPEC-J2) model. The changes of ACE2 mRNA and protein expression levels after PEDV infection in IPEC-J2 were detected by RT-qPCR and Western blot. And the PEDV replication levels were detected by RT-qPCR, Western blot, and TCID₅₀ after overexpression and inhibition of ACE2 expression. The results showed that the mRNA and protein expression levels of ACE2 were significantly downregulated after PEDV infection with IPEC-J2, which was consistent with the transcriptomic results. PEDV infection significantly increased in the ACE2 overexpression group and significantly decreased in the PEDV infection in the ACE2 suppression group. This study validated the role of ACE2 in the process of PEDV infection at the cellular level. PEDV infection was able to decrease ACE2 expression, overexpression of ACE2 in IPEC-J2 cells increased PEDV replication levels, and inhibition of ACE2 expression decreased PEDV replication levels.

Key words: angiotensin converting enzyme 2, ACE2, porcine epidemic diarrhea virus, IPEC-J2

中图分类号:

S855.3

任莉鑫, 张静怡, 徐沙沙, 杨柳, 张兴翠, 宋振辉. ACE2对猪流行性腹泻病毒体外感染传代猪小肠上皮细胞的影响[J]. 畜牧兽医学报, 2024, 55(3): 1238-1248.

REN Lixin, ZHANG Jingyi, XU Shasha, YANG Liu, ZHANG Xingcui, SONG Zhenhui. Effect of ACE2 on Porcine Intestinal Epithelial Cells Infected with Porcine Epidemic Diarrhea Virus in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(3): 1238-1248.

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ACE2对猪流行性腹泻病毒体外感染传代猪小肠上皮细胞的影响

Effect of ACE2 on Porcine Intestinal Epithelial Cells Infected with Porcine Epidemic Diarrhea Virus in vitro

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