白藜芦醇对轮状病毒感染猪肠上皮细胞IPEC-J2的抑制效应  

Inhibitory Effect of Resveratrol on Rotavirus-infected Porcine Intestinal Epithelial Cells IPEC-J2

作　　者：彭宁 梁雅旭 龙菲 余东明 钟翔[1] PENG Ning;LIANG Yaxu;LONG Fei;YU Dongming;ZHONG Xiang(College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物科技学院,南京210095

出　　处：《畜牧兽医学报》2024年第9期4213-4225,共13页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家重点研发计划(2022YFD130040307);中央高校基本科研业务费专项资金——中药全产业链创新研究(KYCXJC2023003)。

摘　　要：旨在研究白藜芦醇对轮状病毒(porcine rotavirus,PoRV)感染猪肠上皮细胞的抗病毒作用。本试验以猪肠上皮细胞(IPEC-J2)为研究对象,先测定不同白藜芦醇浓度对IPEC-J2细胞活力的影响,再根据是否用PoRV或白藜芦醇(100μmol·L^(-1))处理IPEC-J2细胞,分别设置阴性对照组、PoRV感染组和不同时间段添加白藜芦醇+PoRV感染组。通过实时荧光定量PCR技术、病毒滴度测定法和蛋白免疫印迹法检测PoRV在IPEC-J2细胞的复制与增殖情况。结果表明:1)在PoRV感染IPEC-J2的后期和全期添加白藜芦醇(100μmol·L^(-1))可显著抑制PoRV的感染和复制(P<0.05);2)与PoRV感染组相比,在PoRV感染IPEC-J2的全期添加白藜芦醇(100μmol·L^(-1))可极显著抑制IPEC-J2细胞上清液中炎症细胞因子IL-1β、IL-10、TNF-α和IFN-β的含量(P<0.01),后期添加白藜芦醇可显著抑制IL-10和IFN-β的含量(P<0.05);后期和全期添加白藜芦醇极显著抑制了IPEC-J2细胞中的免疫相关因子MDA5、RIG-I、TRIF、MAVS、LGALS9、EIF2 AK 2、IRF 9和IFI44L的mRNA相对表达量(P<0.01),极显著抑制了IPEC-J2细胞中可变剪接因子SRPK1、SRPK2、HNRNPC和HNRNPR的mRNA相对表达量(P<0.01);3)与阴性对照组相比,PoRV感染显著促进了IPEC-J2细胞中LGALS 9的5号外显子和EF2AK 2的2号外显子发生外显子跳跃,而在PoRV感染IPEC-J2的后期和全期添加白藜芦醇(100μmol·L^(-1))可显著抑制靶基因发生外显子跳跃。综上证实,本研究添加100μmol·L^(-1)白藜芦醇可抑制PoRV对IPEC-J2细胞的感染和复制,且在PoRV感染后的维持阶段发挥作用,可为预防和治疗仔猪病毒性腹泻提供新的依据和参考。The aim of this study was to investigate the antiviral effect of resveratrol on porcine rotavirus(PoRV)infected porcine intestinal epithelial cells(IPEC-J2).In this experiment,IPEC-J2 cells were used as research object.Firstly,the effect of different concentration of resveratrol on the cell viability of IPEC-J2 was determined.Then experimental groups including negative control group,PoRV-infected group and PoRV-infected group with the addition of resveratrol at different time points were set up depending on whether the IPEC-J2 cells were treated with PoRV or resveratrol(100μmol·L^(-1)).The replication and proliferation of PoRV in IPEC-J2 cells were detected by real-time fluorescence quantitative PCR,viral titer assay and Western blot.The results showed that:1)The addition of resveratrol(100μmol·L^(-1))significantly inhibited PoRV replication and infection at the late and full phases of viral infection of PoRV infection with IPEC-J2(P<0.05);2)Compared with the PoRV-infected group,the addition of resveratrol(100μmol·L^(-1))during the full phase of PoRV-infected IPEC-J2 extremely significantly suppressed the levels of inflammatory cytokines IL-1β,IL-10,TNF-αand IFN-βin the supernatant fluid of IPEC-J2 cells(P<0.01),while significantly suppressed the IL-10 and IFN-βlevels during the late phase(P<0.05);Extremely significantly suppressed the relative expression of immune-related factors MDA5,RIG-I,TRIF,MAVS,LGALS9,EIF2 AK 2,IRF 9 and IFI44 L in IPEC-J2 cells(P<0.01)during late and full phases;Extremely significantly suppressed the relative mRNA expression of variable splicing factors SRPK1,SRPK2,HNRNPC and HNRNPR in IPEC-J2 cells(P<0.01)during late and full phases;3)Compared with the negative control group,PoRV infection significantly promoted exon jumps in exon 5 of LGALS 9 and exon 2 of EF2 AK2 in IPEC-J2 cells,while the addition of resveratrol(100μmol·L^(-1))significantly inhibited the exon skipping of target genes in the late and full phases of PoRV infection IPEC-J2 cells.In conclusion,this study confi

关 键 词：白藜芦醇 猪轮状病毒 抗病毒 可变剪接 

分 类 号：S828.9[农业科学—畜牧学]