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

作　　者：任莉鑫 张静怡[1] 徐沙沙 杨柳[2] 张兴翠[1] 宋振辉 REN Lixin;ZHANG Jingyi;XU Shasha;YANG Liu;ZHANG Xingcui;SONG Zhenhui(College of Veterinary Medicine,Southwest University,Chongqing 402460,China;Chongqing Academy of Animal Husbandry,Chongqing 402460,China;Immunology Research Center,Institute of Medical Research,Southwest University,Chongqing 402460,China)

机构地区：[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

出　　处：《畜牧兽医学报》2024年第3期1238-1248,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：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)。

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。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 TCID 50 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.

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 

分 类 号：S855.3[农业科学—临床兽医学]