Study On The Role Of Poly(I:C) In The Regulation Of JAK2 And VEGF-A Responsing To CSFV Infection

Classical Swine fever is a kind of highly severe infectious disease which is characterized by high fever and diffuse hemorrhage.In this paper,bioinformatics and molecular biology methods were used to analyze and obtain differentially expressed signaling pathway-related genes in the transcriptome database of macrophages responding to different viral strains of CSFV infection.The macrophage model infected by CSFV was established to detect and analyze the response characteristics of specific cytokines such as JAK signaling pathway and VEGF-A.At the same time,A confirmatory study was carried out on the inhibitory effect of Poly(I:C)on the replication of CSFV,and the regulatory effect of Poly(I:C)on the response of JAK2 and VEGF-A to CSFV infection was further discussed.The results provide basic data for further understanding the infection mechanism and antiviral treatment mechanism of CSFV.The research results are as follows:(1)We construct the GO-TREE based on gene trend expression analysis using The RNAseq of macrophages infected by CSFV Shimen strain,C strain and the control group.Then,we find CSFV Shimen strain infection can not only affect cell migration,cell growth,cell proliferation and cell cycle arrest,also change the inflammatory response,NF kappaB regulation,apoptosis regulation and so on.In the GO-TREE,the JAK-STAT pathway was identified as a significant pathway in response to the infection of CSFV Shimen strain.Further the establishment of a macrophage infection model of CSFV Shimen strain was conducted to detect the post-infection response of members of the JAK-STAT by molecular biology.The quantitative PCR results showed that the mRNA expression of JAK2 and STAT4 is significant inhibited in macrophages after the post-infection of CSFV.(2)Differentially expressed gene analysis also showed that VEGF-A was highly expressed in macrophages infected by CSFV.Further molecular biological was carried out.The quantitative PCR,Western blot and confocal analysis results showed that the transcription and protein expression levels of VEGF-A in macrophages were significantly up-regulated after the infection by CSFV Shimen strain.(3)Conduct quantitative PCR detection to analysis the influence of Poly(I:C)in different concentrations and different time on CSFV Shimen straininfection.The results showed Poly(I:C)has significant inhibitory effect on CSFV replication in macrophages.Immunity fluorescence detection further support this conclusion.GO-TREE analysis suggested that JAK-STAT might be used as an antiviral target of myoblast,and quantitative PCR analysis showed that Poly(I:C)could reverse the immune inhibition of JAK2 and STAT4 induced by CSFV Shimen strain in macrophages,suggesting that Poly(I:C)might play an antiviral role by activating the JAK-STAT pathway.At the same time,the results showed that Poly(I:C)could inhibit the up-regulated expression of VEGF-A in macrophages induced by CSFV Shimen strain,suggesting that Poly(I:C)might alleviate the inflammatory response induced by CSFV by inhibiting virus replication.