Transcriptome Analysis Of PRRSV-Infected MARC-145 And The Inhibitory Effect Of Mg132 On PRRSV Replication

Porcine reproductive and respiratory syndrome(PRRS)caused by porcine reproductive and respiratory syndrome virus(PRRSV)is an swine acute infectious disease.This disease is characterized by reproductive disorders in pregnant sows and respiratory distress in piglets,resulting in significant economic losses in the pig industry worldwide.So far,there is no very effective method of prevention and control.In this study,the transcriptome of PRRSVinfected MARC-145 was analysized.Inhibitroy effect of proteasome inhibitors on PRRSV replication was investigated.It contributes to find a new and more effective method to prevent PRRS.1.The transcriptome analysis PRRSV-infected MARC-145 In this study,the Illumina Nova Seq platform and bioinformatics was employed to analyze a differentially expressed genes(DEGs)of PRRSV-infected MARC-145.The results showed that there were 191 differential expression genes in PRRSV-infected MARC-145.Of them,119 genes demonstrated significantly up-regulated and 72 genes demonstrated significantly downregulated.KEGG analyses indicated that these DEGs enriched in multiple signaling pathways,such as cytosolicDNA sensing pathway,RIG-I-like receptor signaling pathway,TNF signaling pathway,NOD-like receptor signaling pathway,chemokine signaling pathway,Toll-like receptor signaling pathway,Jak-STAT signaling pathway,NF-kappa B signaling pathway,apoptosis,AMPK signaling pathway,cytokine-cytokine receptor interaction and others.Some immune-related genes such as IL-6,IL8/cxcl8,IL20 rb,inhbe,pyrin/mefv,cd30/tnfrsf8,gpcr/chrm5 showed down-regulation,which the immunosuppression was suggested after PRRSV infection in MARC-145.Meanwhile ube2 c gene(in the ubiquitin-proteasome pathway)was significantly up-regulated.Taken together,it indicated that DEGs of the PRRSV-infected MARC-145 involved multiple signaling pathways,the ubiquitin-proteasome pathway and immunosuppression caused by PRRSV infection.2.Inhibitory effect of Mg132 on PRRSV replication In this study,based on evaluation of the cell cytotoxic effect of Mg132 and lactacystin toward MARC-145 by MTT assay firstly,the inhibitory effects of Mg132 and lactacystin on PRRSV replication in vitro using qRT-PCR.The transcriptome of PRRSV-infected MARC-145 after treatment of Mg132 was analyzed using the Illumina Nova Seq platform and bioinformatics.The results showed that Mg132 and lactacystin displayed the inhibitory effect on the replication of PRRSV.And Mg132 was stronger the inhibitory effect than lactacystin.The results of transcriptome analysis showed that there 1691 DEGs in PRRSV-infected MARC-145 after teatment of Mg132.Of them,3114 genes were significantly up-regulated and 3077 genes were significantly down-regulated.KEGG analyses showed that multiple signaling pathways involved this pharmacological process,including: cytosolicDNA sensing pathway(14 DEGs),RIG-I-like receptor signaling pathway(18 DEGs),TNF signaling pathway(47 DEGs),NOD-like receptor signaling pathway(57 DEGs),Chemokine signaling pathway(45 DEGs),Toll-like receptor signaling pathway(39 DEGs),Jak-STAT signaling pathway(39 DEGs),NF-kappa B signaling pathway(28 DEGs),apoptosis(57 DEGs),AMPK signaling pathway(35 DEGs)and Cytokinecytokine receptor interaction(63 DEGs).Many antiviral genes(usp18,oas1,oas3,isg15,isg20,ddit3,sumo3)showed up-regulation.There were 37 DEGs about ubiquitin-proteasome system.Of them,15 DEGs(rhobtb1,map3k1,brca1,cdc20,ubr5,trim37,ube2 q,birc3,skp2,ube2q2,cul1,cul4 b,fbxw7,klhl13,ube2c)showed down-regulation.From above,the pharmacological process on PRRSV-infected MARC-145 after Mg132 treatment involved multiple signaling pathways,up-regulation of some antiviral gene expression,and downregulation of some gene expression about ubiquitin-proteasomesystem.The mechanisms on antivirus of Mg132 was complex.In summary,it was found that pathological mechanisms of PRRSV-infected MARC-145 involved many signaling pathways,antivirus gene and genes about ubiquitin-proteasome pathway.And slso antivirus of Mg132 involved many signaling pathways,antiviral genes,and ubiquitin-proteasome pathway.