Dynamic Change Analysis Of P53-related LncRNAs Levels In Spleen And Feather Pulp Of MDV-infected Chickens

Marek’s Disease virus(MDV),a highly oncogenic herpesvirus associated with cells,is the pathogen of Marek’s disease(MD).MDV infection can lead to a lymphoproliferative disease in poultry and lead to immunosuppression in chickens.Although the use of MD vaccine can effectively prevent and control the occurrence and spread of the disease,when the prevention and control measures are not in place or the virulence of MD virus changes,the disease still causes serious harm to the poultry industry and brings huge economic losses.Therefore,the prevention and control of the spread of MD remains a challenge for the poultry industry.Surveillance of chicken virus infection is an important link in MD prevention and control,and screening of markers for MDV infection surveillance is one of the key issues.In recent years,the RNA strand length is more than 200 nucleotides noncoding RNA(lnc RNA),participate in a variety of disease control process,and the course of disease development,through the way such as regulating the disease related gene expression level,in the process of virus infection disease occurrence and development plays an important role,at the same time,as a new disease marker for diagnosis and prevention and control targets,also attracted much attention.In our previous research,five differentially expressed lnc RNAs related to the tumor-related protein p53 were obtained through RNA-seq prediction analysis.They are respectively p53 binding protein 1lnc RNA-1(TP53BP1lnc RNA-1),p53 binding protein 1lnc RNA-3(TP53BP2Lnc RNA),p53 target gene 5Lnc RNA(TP53TG5Lnc RNA),and p53 inducing gene 3Lnc RNA(TP53I3Lnc RNA).In this study,the above five p53-related lnc RNAs were dynamically analyzed in the spleen and feather pulp tissues of chickens infected with MDV at different stages,aiming to provide reference for the screening of potential markers for early monitoring of MDV infection in chickens and the development of targeted drugs.In this study,SPF chickens were randomly divided into two groups,which were isolated and raised under the same conditions.The experimental group was intraperitoneally injected with 0.5 m L BJ-1MDV0.5m L/chicken(1000PFU/per individual),while the control group was injected with the same amount of PBS(p H=7.4),and spleen tissues and feather pulp medulla tissues of chicken were collected at 1,7,14,21,35 and 42 d after infection.The transcriptional levels and protein expression levels of five p53-related lnc RNAs in chicken spleen and feather pulp tissues were detected and analyzed by q PCR and Western Blot.The results showed that :(1)compared with the control group,the transcription levels of four lnc RNAs(TP53BP1lnc RNA-1,TP53BP1 lnc RNA-3,TP53BP2 Lnc RNA and TP53TG5 Lnc RNA)in the spleen tissues of chickens infected with MDV at different stages showed trend of decreasing first and then increasing trend with the increase of age,and the differences between groups were significant(P < 0.05).The transcriptional level of 1 lnc RNA,namely TP53I3-lnc RNA,showed a gradually decreasing trend with the increase of age,and the difference between groups was significant(P < 0.05).(2)Compared with the control group,the transcription levels of four lnc RNAs(TP53BP1lnc RNA-1,TP53BP1 lnc RNA-3,TP53BP2 Lnc RNA and TP53TG5 Lnc RNA)in chicken pulp tissues at different stages of MDV infection presented trend of decreasing first and then increasing trend with the increase of age,and the differences between groups were significant(P < 0.05).The transcriptional level of 1 lnc RNA,namely TP53I3-lnc RNA,showed a gradually decreasing trend with the increase of age,and the difference between groups was significant(P < 0.05).The changes of TP53BP1 lnc RNA-1,TP53BP1 lnc RNA-3,TP53BP2 Lnc RNA and TP53TG5 Lnc RNA in the spleen and feather pulp tissues were the same,and showed a gradually increasing trend with the increase of daily age,and the differences between groups were significant(P < 0.05).The variation trend of TP53I3-lnc RNA in spleen and feather pulp was the same,which showed a gradually decreasing trend with the increase of age,and the difference between groups was significant(P < 0.05).The results showed that :(1)compared with the control group,the expression level of p53 protein in MDV-infected chicken spleen tissues showed an upward trend with the increase of infection time,and the difference between the two groups was extremely significant(P < 0.01).(2)Compared with the control group,the expression level of p53 protein in the feather pulp tissue of MDV infected chickens showed an upward trend with the increase of infection time,and the difference between the two groups was extremely significant(P < 0.01).The results of this study showed that after MDV infection,p53-related lnc RNAs showed a consistent up-regulation or down-regulation trend in chicken spleen tissues and feather pulp tissues with the increase of infection time,and p53 protein showed an extremely significant increase trend in spleen tissues and feather pulp tissues.The results showed that the p53-related lnc RNA may have a certain effect on the expression and function of the tumor-related protein p53 during the occurrence and development of MD,and thus promote the pathogenesis of MD,providing a reference for the screening of markers for MD detection.The above-mentioned lncrnas in spleen tissue and feather pulp tissue showed consistent changes during the course of MDV infection.Therefore,the non-invasive samples of chicken feather pulp could be used as detection samples for monitoring MDV infection in chickens,thus reducing the damage to chickens and providing convenience for monitoring,detection and diagnosis of MDV infection in poultry flocks.