The Effect Of REV Infection On The Expression Of Apoptosis-related Factors In HD11 And DT40 Cells

With the rapid development of animal breeding industry,disease prevention（control）has become particularly important in recent years.Animals infected with REV happen immunosuppression,it means easy to cause secondary and mixed infections of other pathogens,which makes the diagnosis and prevention of the disease more difficult.The study of the interaction between REV infection and immune cells is helpful to understand the causes of its immunosuppression and tumor formation,clarify its pathogenic mechanism,and provide a theoretical basis for the prevention and treatment of RE.Therefore,according to the characteristics of REV immunosuppression,selected HD11（chicken macrophage cell line）and DT40（chicken lymphoma cells）as the research objects,applied molecular biology technology and immunolog ical methods combined with detection kits were used.mitochondrial membrane potential,Ca²⁺content and apoptosis rate were detected.At the same time,the mitochondrial pathway-related apoptosis factors Caspase-3,Caspase-9,Bax,Bcl-2 expression of mRNA and content of protein were detected.Changes in the main research results are as follows:（1）The virus was detected in HD11 and DT40 cells infected with REV,indicating that REV can replicate and proliferate in HD11 and DT40 cells.And 48 hpi of HD11 cel ls,the viral load reached the peak;REV infection of DT40 cells 12-72hpi load showed an upward trend.（2）The viability of HD11 cells was significantly（P<0.01）lower than the control at 24-36 hpi.The viability of DT40 cells was significantly（P<0.05）lower than the control at 12 hpi.and extremely significantly（P<0.01）lower than control cells at 48 hpi.（3）The mitochondrial membrane potential was significantly（P<0.05）or significantly（P<0.01）lower than that of control cells 36-48 hpi and 36-72 hpi in HD11 and DT40 cells respectively.（4）The Ca²⁺content was significantly（P<0.05）higher at 36 hpi and 48 hpi than the control cells in HD11 and DT40 cells both;although they were also higher than the control cells at other time points,they were not statistically significant（P>0.05）.（5）In HD11 cells,the apoptosis rate was extremely significant（P<0.01）higher than that of control cells without virus infection24-36 hpi and 72 hpi,but the apoptosis rate was higher than that of control cells at 12 hpi,but there was no statistical difference（P>0.05）;In DT40 cells of REV,the apoptosis rate of was extremely significant（P<0.01）than that of control cells at 12、48and 72 hpi.（6）In HD11 cells,Bcl-2 mRNA expression was extremely significantly（P<0.01）lower than control cells in 24-36 hpi,while significantly（P<0.05）or extremely significant（P<0.01）higher than control cells in 48-72 hpi;Bax mRNA expression was significantly（P<0.05）lower than control cells at 24 hpi,and significantly（P<0.05）or very significantly（P<0.01）higher than control cells at 48 hpi and 72 hpi;caspase-3 mRNA expression was extremely significant（P<0.01）higher than control cells at 36 hpi and 72 hpi;caspase-9 mRNA expression was Significantly（P<0.05）or extremely significant（P<0.01）higher than control cells.at 12、36 and 72 hpi;In DT40 cells,Bcl-2 mRNA expression was significantly（P<0.01）higher than control cells at 12hpi and 48-72 hpi;Bax mRNA was not detected during 12-72 hpi（Ct value≥35）;caspase-3 mRNA expression is extremely significantly（P<0.01）higher than control cells at 24 hpi,and significantly（P<0.05）higher than control cells at 36 hpi and 72 hpi;caspase-9 mRNA expression was extremely significant（P<0.01）higher than control cells is at 24-36 hpi,and significantly higher（P<0.05）than control cells at 72 hpi;there was no statistical difference among others（P>0.05）.（7）In HD11 cells,the caspase-3 enzyme activity was extremely significant（P<0.01）higher than control cells at 12-72 hpi;Bcl-2 protein content was extremely significant（P<0.01）Compared with control cells at 24 and 72 hpi,the content of caspase-9 protein was significantly（P<0.01）higher at 12-48 hpi.In DT40 cells,its caspase-3 enzyme activity was significantly（P<0.01 or P<0.05）higher than control cells at 36-48 hpi and 72 hpi;Bcl-2 protein content was extremely significant（P<0.01）higher than control cells at 12 and 48 hpi,but significantly（P<0.01）lower than control cells at 72 hpi;caspase-9 protein content was extremely significant（P<0.01）higher than control cells at 12-24 hpi,while significantly（P<0.01）lower than control cells at 48 hpi.no statistical difference was found in the rest（P>0.05）.Through the detection of the above indicators,the purpose is to explore the function a nd role of the mitochondrial apoptosis pathway in the process of REV infection of cells,to provide an in-depth explanation of the immune suppression mechanism of REV and the role of mitochondrial apoptosis pathway in the apoptosis caused by viral infectio n New theoretical basis.