Effect Of Acanthopanax Senticosus Extract On Porcine Pseudorabies Virus And Pig Immune Function

This study systematically explored the anti-PRV virus activity and anti-viral mechanism of Acanthopanax senticosus extract through in vitro cell experiments and in vivo animal experiments.In order to study the effect of Acanthopanax senticosus extract on the proliferation of porcine pseudorabies virus(PRV)on PK-15 cells,PK-15 cells were infected with PRV,and different concentration gradients of 1.5 mg/m L,1 mg/m L,0.75 mg/m L,0.5 mg/m L,0.25mg/m L,results show that:1)Acanthopanax senticosus extract has a certain inhibitory effect on PRV virus in a dose and time-dependent manner.When the concentration range of Acanthopanax senticosus extract is 1.5-2.0 mg/m L,the antiviral effect of Acanthopanax senticosus extract is optimal with highest safety and antiviral effect;2)After PRV was applied to PK-15 cells for 3 hours,the inhibitory effect of Acanthopanax senticosus extract at 1 mg/m L on PRV was the best.In summary,our results demonstrated that Acanthopanax senticosus extract can inhibit PRV proliferation in vitro.In order to study the effect of Acanthopanax senticosus extract on the virus proliferation,histopathological effect and inflammatory factor levels in artificially infected PRV mice.Forty female ICR mice were randomly divided into 4 experimental groups(I,?,III,and IV);and 10 mice/group.Group?was a blank control group(fed with common feed,not challenged),group?was challenged with a control group(fed with ordinary feed,challenged),and group III was a control group(fed with acanthopanax extract,Not challenged),group IV was the experimental group(fed with Acanthopanax senticosus extract feed,challenged),the results showed:1)4mg/g before challenge for 5 days,when developed symptoms the mice were sacrificed,and it was found that the administration group could significantly inhibit the proliferation of PRV in inguinal lymph nodes and lung tissues(P<0.05);2)Pathological sections and immunohistochemical results.Acanthopanax senticosus extract can reduce the pathological damage of brain and lung tissue caused by PRV;3)Cytokine test showed that there were significant differences in MCP 1,MIP-1?,IL1-?,IL1-?between the drug-free group and the drug-infected group in brain tissue(P<0.05).The expression of inflammatory cytokines was significantly up-regulated compared with the untreated group(P<0.05);MCP-1,MIP-1,G-CSF are important cytokines for monocyte aggregation and maturation,and are the key to promote non-specific immune activation;IL1-?,IL1-?and IL2 are important cytokines that promote Th1-type cellular immunity;the up-regulation of these cytokines indicates that the mice fed with Acanthopanax senticosus extract showed a more rapid and effective immune response to PRV infection.The above results indicate that the extract of Acanthopanax senticosus can significantly improve the immune function of mice,inhibit the colonization and pathogenicity of PRV in mice.Another 10 female ICR mice were randomly divided into 2 groups,the experimental group(fed with Acanthopanax senticosus feed)and the control group(fed with ordinary feed).After continuous feeding for 10 days,80 and Transcription levels of innate and adaptive immune-related genes were detected.The results showed that there were no significant differences in the transcription levels of the 52 genes between the medicated and non-medicated groups.The 7 genes were significantly higher in the medicated group than in the unmedicated group,and the transcription levels of 21 genes were significantly lower in the medicated group.In order to study the effect of Acanthopanax senticosus extract on growth performance and immune regulation of piglets,36 piglets(12 sows each)of 3 sows were grouped.The 12piglets of each sow were divided into 4 groups and fed with 0g/pig,2g/pig,4g/pig,6g/pig Acanthopanax extract diet.The piglets were weighed to determine growth indicators before and after the experiment,and blood samples were taken to determine blood indicators and cytokines.The experimental results show that:1)Adding 2g/pig and 4g/pig Acanthopanax extract to the diet can significantly increase the growth rate of piglets and increase the feed utilization rate(P<0.05);2)Differential counts of peripheral blood showed that the addition of 2g/pig,4g/pig and 6g/pig to diet can significantly increase the number of white blood cells and lymphocytes in piglets(P<0.05);3)Cytokine ELISA test.Adding 6g/pig of Acanthopanax senticosus extract to the diet significantly increased the IFN-?content in the serum of piglets(P<0.05).One piglet from each group was selected.A total of 12 piglets were slaughtered andspleen was extracted.After RNA was extracted,the transcriptome was sequenced and analyzed,and the sequencing results were verified by q RT-PCR.Western blot was used to verify the protein expression levels of immune-related pathways.The results showed that:1)in the 6g group and the control group,the transcripts in the group were well correlated(R~2>0.8);2)the accuracy of the sequencing results was confirmed by q RT-PCR;3)the Western blot results showed that the relative protein expression levels were consistent with the relative transcription levels of genes in the transcriptome data,that is,compared with the control group(0 g),the transcription and expression of NF?B1 were slightly up-regulated in the experimental group(2,4,6 g),and the transcription and expression of TBX21 and ZAP70 were significantly up-regulated.In summary,this study conducted a study and analysis of the effects and effects of Acanthopanax senticosus extract on PRV infection in vitro and in vivo.The results showed that Acanthopanax senticosus extract had a good inhibitory effect on the proliferation of PRV in cells and mice,and up-regulated growth performance and immune-related factors in piglets.The research results provide experimental basis for further digging the antiviral mechanism of Acanthopanax senticosus extract and clinical prevention and control of PRV.