Duck Enteritis Virus Induces Endoplasmic Reticulum Stress-Mediated Autophagy And Apoptosis Of Duck Embryo Fibroblasts And Its Related Mechanism

Duck virus enteritis?DVE?,also known as duck plague?DP?,is an acute,contagious disease caused by duck enteritis virus?DEV?.It is characterized by high mortality,and bring huge economic losses to the duck industry.DEV,also known as Duck plague virus?DPV?,belongs to the herpes virus family,the eighth virus classification report classified it as a herpesviridae unclassified virus.In recent years,some studies have shown that DEV can cause autophagy in duck embryo fibroblast?DEF?cells,and highly infectious viruses can cause DEF cell death by affecting intracellular Ca²⁺imbalance.In addition,DEV can induce autophagy by responding to endoplasmic reticulum stress related unfolded proteins.At present,researchers focus their research on DEV molecular biology,however,there are few studies on cell function and related mechanisms after infection of DEV in related cells.In this study,This study takes autophagy and endoplasmic reticulum stress as the research objects,to provide relevant theoretical basis for elucidating the mechanism of DEF infection DEF,and provide relevant ideas for the prevention and control of duck plague..The main research contents are as follows:1.The effect of DEVs with different multiplicity of infection on DEF proliferation and apoptosis:DEF was infected with 0.1 MOI,1 MOI and 10 MOI,CCK8 was used to detect cell proliferation,and flow cytometry was used to detect apoptosis.The results showed that the higher the DEV concentration,the lower the cell viability and the higher the apoptosis.1 MOI DEV infected DEF,the cell viability decreased significantly,and 10 MOI DEV infected DEF,the cell viability dropped to below 50%.2.The effect of DEV of different MOI on Ca²⁺level,autophagy and ER stress related protein levels in DEF:DEF was infected with 0.1MOI,1MOI and 10MOI DEV,fluorescent probe Fluo-4 was used to detect Ca²⁺levels,WB was used to detect autophagy-related proteins LC3-I,LC3-II,endoplasmic reticulum stress-related proteins CHOP,GRP78 and ATF6 expression levels.The results showed that DEV induces Ca²⁺levels and up-regulates the expression of LC3-II/LC3-I,while the expression levels of CHOP,GRP78 and ATF6 are up-regulated,and has a dose-response relationship with the concentration of DEV.3.Ca²⁺chelating agent reverses the effect of DEV on DEF infection:After infecting DEF with 10MOI of DEV,add 10m M Ca²⁺chelating agent?EDTA?,fluorescent probe Fluo-4 to detect Ca²⁺level,CCK8 to detect cell proliferation,flow cytometry to detect apoptosis,WB to detect autophagy-related protein LC3-I,LC3-II,expression levels of endoplasmic reticulum stress-related proteins CHOP,GRP78 and ATF6.The results showed that Ca²⁺chelating agent can significantly promote DEV-mediated DEF proliferation activity and reduce DEV-induced apoptosis.At the same time,Ca²⁺chelating agent can reduce the expression of LC3-II/LC3-I and the expression of CHOP,GRP78 and ATF6.Conclusion:DEV inhibits the proliferation of DEF,promotes the apoptosis of DEF,promotes the flow of extracellular calcium ions,and induces endoplasmic reticulum stress and autophagy in DEF in a concentration-dependent manner;while Ca²⁺chelator can reverse the effect of DEV on DEF infection.