The Effect Of ATF3 On Cell Cycle And Apoptosis In The N2a Cells Infected With Porcine Hemagglutinating Encephalomyelitis Virus

Porcine hemagglutinating encephalomyelitis is caused by porcine hemagglutinating encephalomyelitis virus,mainly infected 3 weeks old piglets,clinical manifestations of vomiting failure and obvious neurological symptoms.In order to find out whether ATF3 has high expression of ATF3 and its high expression mechanism after ATV3 infection in human neurons,the effect of ATF3 on the expression of ATF3 before and after PHEV was observed in this study.The results showed that ATF3 was highly expressed in mice after PHEV infection.And to explore the effect of JNK signaling pathway on ATF3 expression in porcine hemagglutinating encephalomyelitis virus infected neurons.N2a cells were used as the study model.After culturing N2a cells into the monolayer in 6-well plates,the total RNA and total protein were extracted at 2h,12h and 24h after inoculation.PCR and Western blot were used to detect the expression of ATF3 mRNA and protein.According to the test requirements,the blank control group,the poisoning group,the DMSO group,the DMSO poisoning group,the JNK inhibitor group and the JNK inhibitor poisoning group.The total RNA and total protein were extracted at 2h,12h and 24h,respectively.The total RNA was extracted by fluorescence quantitative PCR and Western blot(Western Blot),and the total RNA was extracted from the two groups.The mRNA and protein expression of ATF3 were measured respectively.After the same grouping,the apoptosis and cell cycle of 12h and 24h were detected by Annexin V-FITC/PI double staining and flow cytometry.The expression of ATF3 gene in normal cells was at steady and low level,and the expression of ATF3 gene was decreased or increased with the decrease of PHEV gene expression.The trend of ATF3 gene was consistent and positively correlated with PHEV.The expression of ATF3 was significantly decreased after treated with JNK inhibitor to N2a cells.By analyzing the results of flow cytometry,we found that the apoptosis rate of JNK pathway was significantly higher than that of the control group(p<0.01),which indicated that ATF3 could inhibit the apoptosis in JNK pathway,And the cell cycle of J-NK inhibitor group was blocked,DNA replication and synthesis were affected,resulting in decreased cell viability,decreased antiviral ability,increased apoptosis rate.The expression of ATF3 was increased after N2a cells infected with PHEV.After JNK inhibitor on N2a cells,ATF3 expression decreased,the apoptosis rate increased,cell cycle arrest,DNA synthesis blocked,decreased cell viability,anti-virus ability decreased.Indicating that JNK signaling pathway can regulate the expression of ATF3 after neurotransmitter infection in PHEV,which in turn affects neuronal apoptosis and cell cycle.