| Japanese encephalitis virus(JEV)infection induces brain tissue diseasecharac-terized by neuron damage and gliosis.Outbreak of Japanese encephalitis comes with high mortality rate and serious sequela.However,the potential mechanism remains largelyunknown and there still has no specific therapeutic drug toward Japanese encephalitis.Therefore,research on molecular mechanism and pathogenicity of apoptosis induced by JEVwillprovide new insights and targets for the development of special anti-JE drugs.Endoplasmic reticulun stress response(ERS)is a signal transduction pathway that triggered by cell-inside or outside factors or overload of misfolded or unfolded proteins.ERS is involved in a series of diseases,such as nervous system diseases,hepatic diseases and diabetes.Early study of our lab indicated that JEV could trigger BHK-21 cell death via inducing IRE1/JNK signaling pathway of ERS.To reveal the mechanism of JE in protein level,we had profiled gene expression of Mouse Brain and Neuro-2a infected with JEV.Integration analysis of in vitro and in vivo mRNA transcriptome revealed that ERS was both induced and HSP70 was both significantly up-regulated.Furrhermore,RNA-sequence indicated that JEV infection regulated multiple signaling pathways such as steroid biosynthesis,p53 signaling pathway,TNF signaling pathway,Foxo signaling pathway,MAPK signaling pathway and ECM-receptor interaction.Among all biological functions that regulated by Foxo protein,cell cycle,aging and apoptosis rank the most important ones.JEV infection induce apoptosis of Neuro-2a cells.Interestingly,after 48 h treatment in Neuro-2a,transciptome analysis,real time PCR and immunoblot analysis found out that JEV infection decreased the expression and activity of Foxo and it’s downstream gene Bim which indicated that Foxo1 induce cell apoptosis may not through regulating its downstream gene Bim.In order to further identify the role of Foxo1 playing in regulating apoptosis of Neuro-2a infected with JEV,we carried out the research via RNA silence and over expression technique.We found out that RNA silence of Foxo1 significantly decrease translation level of Foxo1 and Bim which resulted in higher apoptosis rate of Neuro-2a.On the contrary,over expression of Foxo1 decreased apoptosis rate of Neuro-2a via increasing expression of Foxo1 and Bim.Therefore,we identified for the first time that Foxo1 could inhibit apoptosis of Neuro-2a in response to JEV infection but not through regulating its downstream gene Bim. |
