Zika Virus Inhibits Interferon Production By Phosphorylating AMPK

Background:zika virus(Zika virus,ZIKV)belongs to flavivirus,caused by infectious diseases mainly through Aedes mosquitoes,caused a huge burden to the world public health security,caused by ZIKV infection for unspecific clinical manifestations,can not cause people attention,but also can cause serious neurological diseases such as microcephaly,gill-barre syndrome,become a global concern of public health issues.Interferon(IFN)plays a key role in resisting viral infection.Host cells after from ZIKV infection,this Retinoic acid-inducible gene I-induced gene protein Ⅰ(RIG-Ⅰ)recognizes infected viral RNA,then binds mitochondrial Mitoehondrial antiviral signaling protein(MAVS)and signals to downstream IFN,but lactate disrupts RIG-I binding to MAVS,leading to RIG-I-like receptors(RIG-I-like receptors,RLRs)Mediated type I interferon production is inhibited.West Nile virus(WNV),Hepatitis C virus(HCV)and Dengue virus(DENV)can upregulate glycolysis in host cells to facilitate the replication of the virus itself,but it is not clear the relationship between ZIKV infection and glycolysis in host cells.So far,the epidemic caused by ZIKV infection is still posing a serious threat to people’s health and safety,and people have not yet been developed to combat ZI The specific agent of KV infection,so studying this drug becomes an urgent task.Our study illustrates the relationship between the glycolytic pathway and the host cell native immunity and provides new ideas for investigating the therapeutic targets of ZIKV infection.Methods:First,measuring the relative lactate yield of ZIKV-infected cells at different time periods and the relative intracellular lactate yield of ZIKV infection under AMPK agonist/inhibitor treatment at a specific time.Second,two sets of cell culture media with different glucose concentrations were set,and the relative yield of IFN-under ZIKV infection was determined by fluorescence quantitative PCR.Third,three treatment groups were set when infected with or without ZIKV:blank control,glycolysis inhibitor group,and lactate complement group,and the relative transcript level of IFN-was measured by qRT-PCR.Fourth,to determine the sugar For the effect of changes in metabolism on IFN-production,siRNA was used to reduce the expression of key enzymes in glucose metabolism,IFN-3-glyceraldehyde phosphate dehydrogenase(GAPDH)transcript level was measured by qRT-PCR,and HK2 and GLUT1 and phosphorylation of AMPK were measured by western-blot.Results:After ZIKV infection,intracellular lactate production increased with infection.Inhibition of the glycolytic pathway,reducing lactate production,promotes cells to produce more IFN-after infection with ZIKV.After infection with host cell HEK293T,Zika virus promotes phosphorylation of intracellular AMPK and exerts protein kinase activity,resulting in upregulation of intracellular glycolysis levels to produce more lactate,which ultimately suppresses IFN-production mediated by intracellular RLR signaling.Conclusions:We demonstrated that ZIKV activates the Adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK)after infection with HEK293T cells.Then,the adoption of AICAR(Acadesine)activated AMPK,resulting in increased ZIKV replication,and dihydrodeoxymorphine,also known as compound C(Compound C),inhibited AMPK activation,resulting in reduced ZIKV replication.Second,AMPK activation also enhances the expression of Hexokinase2(HK2)and Glucose transporter-1(GLUT1),which both induce the upregulation of intracellular glycolysis.In addition,we used various methods to alter the activity of glucose metabolism,and we found that upregulation of the Tricarboxylic acid cycle(TCA)pathway or inhibiting the glycolytic pathway led to a reduction in lactate production,ultimately reducing IFN production and promoting ZIK Replication of V.