ZnSO_(4)通过干扰PRRSV的生命周期阶段抑制PRRSV的增殖  

Zinc sulfate inhibits PRRSV proliferation by interfering with different stages of PRRSV lifecycle

作　　者：杨惠珍 李晓姣 孙家振 吴子璇 范阔海[1] 尹伟[1] 孙娜[1] 张振彪 李宏全[1] Yang Huizhen;Li Xiaojiao;Sun Jiazhen;Wu Zixuan;Fan Kuohai;Yin Wei;Sun Na;Zhang Zhenbiao;Li Hongquan(Shanxi Key Laboratory for Modernization of TCVM/College of Veterinary Medicine,Shanxi Agricultural University,Jinzhong 030801,China)

机构地区：[1]山西农业大学动物医学学院/中兽医药现代化山西省重点实验室,山西晋中030801

出　　处：《山西农业大学学报（自然科学版）》2024年第4期51-59,共9页Journal of Shanxi Agricultural University(Natural Science Edition)

基　　金：山西省基础研究计划项目(20210302124138);山西省科技创新人才团队专项(202204051001021);山西农业大学科技创新基金博士科研启动项目(2020BQ66);山西农业大学大学生创新创业训练计划项目(S202410113049)。

摘　　要：[目的]本文旨在探究硫酸锌(Zinc sulfate,ZnSO_(4))抑制猪繁殖与呼吸综合征病毒(Porcine respiratory and re⁃productive syndrome virus,PRRSV)复制的作用,明确Zn^(2+)载体槲皮素(Quercetin,QR)增强Zn抗PRRSV感染的作用,为进一步将ZnSO_(4)联合QR作为潜在预防和治疗PRRSV感染的有效策略提供理论支持。[方法]利用细胞培养半数感染量(Cell culture infective dose 50%,CCID_(50))的方法检测ZnSO_(4)和ZnCl_(2)对PRRSV滴度的影响。利用实时荧光定量PCR(Quantitative Real-time PCR,qRT-PCR)和蛋白印迹(Western blot,WB)检测ZnSO_(4)处理Marc-145和PAMs细胞后,PRRSV N基因和N蛋白表达量的变化。利用qRT-PCR检测ZnSO_(4)预处理24 h后,在4℃和37℃处理下PRRSV N基因表达量的变化。同时,利用qRT-PCR检测ZnSO_(4)处理不同时间,培养基上清中PRRSV N基因表达量的变化。利用qRT-PCR和WB检测ZnSO_(4)、QR以及两者联合处理,PRRSV N基因和N蛋白表达量的变化。[结果]CCID_(50)结果表明:ZnSO_(4)和ZnCl2均可显著降低PRRSV滴度。在Marc-145和PAMs上,不同浓度ZnSO_(4)处理,均可显著抑制PRRSV N基因和N蛋白表达,且抑制作用呈明显的剂量-效应关系。不同浓度ZnSO_(4)预处理24 h,在4℃和37℃处理2 h后,均可显著降低PRRSV N基因表达,且呈明显的剂量-效应关系。100μmol·L^(–1)ZnSO_(4)处理,在不同时间段,可显著抑制上清中PRRSV N基因的表达。ZnSO_(4)联合QR处理结果表明,ZnSO_(4)联合QR可显著抑制PRRSV N基因和N蛋白的表达。[结论]ZnSO_(4)能显著抑制PRRSV的复制,它不仅可以阻止PRRSV的黏附和进入,而且可以抑制PRRSV的复制和干扰PRRSV的释放。此外,QR可以增强ZnSO_(4)抗PRRSV感染的作用。本研究为进一步明确ZnSO_(4)抗PRRSV感染的分子机制奠定了理论基础。[Objective]This study mainly aimed to investigate the role of ZnSO_(4)in inhibiting the replication of porcine respiratory and reproductive syndrome virus(PRRSV),clarify the enhancement of Zn’s antiviral effect against PRRSV by Zn^(2+)carrier quercetin(QR),and provide theoretical support for using ZnSO_(4)combined with QR as a potential effective strategy for preventing and treating PRRSV infection.[Methods]The effects of ZnSO_(4)and ZnCl_(2)on PRRSV titers were detected using the cell culture infective dose 50%(CCID 50)method.Quantitative Real-Time PCR(qRT-PCR)and Western Blot(WB)were used to determine changes in PRRSV N gene and N protein expression in ZnSO_(4)-treated Marc-145 and PAMs cells.The expression of the PRRSV N gene was measured using qRT-PCR after pretreatment with ZnSO_(4)for 24 hours at 4℃and 37℃.Additionally,the changes in PRRSV N gene expression in the culture supernatant at different treatment time points following ZnSO_(4)treatment were also measured using qRT-PCR.The expression changes of PRRSV N gene and N protein with ZnSO_(4),QR alone,and in combination were detected using qRT-PCR and WB.[Results]CCID 50 results indicated that both ZnSO_(4)and ZnCl_(2) significantly reduced PRRSV titers.In Marc-145 and PAMs cells,different concentrations of ZnSO_(4)significantly inhibit the expression of PRRSV N gene and N protein in a dose-dependent manner.After 24 hours of pretreatment with different concentrations of ZnSO_(4),the expression of PRRSV N gene was significantly reduced following two hours of treatment at 4℃and 37℃,also showing a dose-dependent relationship.Treatment with 100μmol·L^(–1)ZnSO_(4)significantly inhibited the expression of PRRSV N gene at different time points.Results from the combined treatment of ZnSO_(4)and QR indicated that the combination significantly inhibited the expression of the PRRSV N gene and N protein.[Conclusions]ZnSO_(4)significantly inhibited PRRSV replication by preventing the virus’s attachment and entry,inhibiting replication,and interfering wi

关 键 词：硫酸锌 猪繁殖与呼吸综合征病毒 抗病毒 槲皮素 

分 类 号：S852[农业科学—基础兽医学]