Study On Porcine Parvovirus Regulate Expression Of Inflammatory Factor Through Activation Of NF-?B Signal Pathway

Porcine parvovirus(PPV)is one of the main pathogens of reproductive barriers;which have caused serious damage in China even all over the world to the breeding of pigs.PPV is mainly transmitted through respiratory tract,the digestive tract and the placenta.Sows,boars and persistent infection health pig all can spread the disease.PPV also co-infected with other virus,such as Japanese encephalitis virus(HEV)and Porcine circovirus(PCV),and this makes things worse.Due to the widespread popularity and harm of this disease,it has been one of the research focuses.Today,the research about PPV often focuses on the method of PPV detection and the vaccine,but mechanism of how PPV infected cells is not clear.The NF-?B signal pathway is one important signal pathway.It has been confirmed that NF-?B participates in the process of many kinds of virus infection,such as H-1 small virus,parvovirus B19.But if the Porcine parvovirus infection cab activate the NF-?B pathway?And through the pathway to regulate the body's natural immune response.Therefore,further study on the mechanism of how PPV activate the NF-?B signaling pathway is important.It not only has important significance to clarify PPV infection mechanism,but also can enrich the NF-?B signaling pathway to virus activation mechanism.According to the results of our previous studies,ater infected PK-15 cell,PPV can cause a series of changes in cell inflammatory factor in the m RNA level.We conclude that PPV can be through the NF-?B signaling pathway regulate the expression of inflammatory cell factors.In our experiments,combining the role of NF-?B pathway in viral infection,we are gonging to research the relationship between PPV and NF-?B pathway.Our purpose is to clarify the relationship between PPV and NF-?B pathway,as well as the change of inflammatory expression witch mediated by NF-?B signal pathway.Initially,after PPV infected PK-15 cells,the transcription phases about 40 kinds of proteins related to NF-?B signal pathway were detected by real time PCR.And the results showed that there is a number of transcription phases about proteins changed obviously,and the same to TLR7/9.These results support our inference,in a certain extent.PPV can regulate expression of inflammatory through the NF-?B signal pathway.Then,we use different experiments to study the influence on NF-?B signal pathway activation after PK-15 cells infected with PPV.We use the luciferase dual reporting system to detect the ability of NF-?B binding to DNA in PK-15 cells infected with PPV.The results showed that PPV can obviously enhance the binding ability of NF-?B,and it is a dose dependent.But the UV inactivated PPV don't have this ability.We detect the influence on p65 nuclear transfer situation after PPV inected PK-15 cells,and the influence on the degradation of I kappa alpha B situation.All of the results showed that PPV can acculate activation of NF-?B signal pathway.Lastly,we detect the change of inflammatory cytokines expression by ELISA,IL-6/IL-12/TNF-?,after PPV infection PK-15 cells,as well as the relationship between the change and NF-?B signal pathway.The ELISA results shoed that the expression level of IL-6/IL-12/TNF-? changed obviously,after PPV infection PK-15 cells.We also use NF-?B signal pathway inhibitors BAY11-7082 to stimulate PK-15 cells,after PK-15 infected with PPV,then detecte the express level of these cytokines again by ELISA.The results showed that the expression level of these cytokines change in the opposite compared with the group there is no BAY11-7082.From what has been discussed above,we get such a conclusion,PPV infecting PK-15 cells can activate the NF-?B pathway,and can regulate the expression of downstream immune factors through this channel.