IBDV感染对法氏囊RNA m^6A和DNA 5mC修饰以及相关催化酶基因表达的影响

Effects of IBDV infection on bursal 5mC,m^6A and the gene expression of associated enzymes

传染性法氏囊病毒(IBDV)感染造成病鸡法氏囊严重损伤,引起免疫抑制,然其具体机制仍不明晰。本研究旨在探讨IBDV感染对法氏囊表遗传修饰及其相关酶表达的影响。试验采用21日龄商品肉鸡,给予IBDV处理,感染后3,5 d收集法氏囊组织。应用基于ELISA试剂盒检测总5mC和m^6A,Real-time PCR检测相关催化酶基因表达。结果显示,感染后5 d,总m^6A水平及其甲基转移酶METTL3和METTL14 mRNA表达均显著降低(P<0.01);去甲基化酶FTO的表达在感染后3,5 d均显著降低(P<0.05)。总5mC水平在感染后3 d明显升高(P<0.05),其去甲基化酶Tet1 mRNA表达明显降低(P<0.01);感染后5 d,甲基转移酶DNMT1 mRNA表达显著降低(P<0.01),而DNMT3a的表达明显升高(P<0.01);结果表明IBDV能够影响法氏囊5mC和m^6A谱及相关酶的表达,这可能是IBDV影响宿主免疫功能的途径之一。

Infectious bursal disease virus(IBDV)infection causes excessive bursal inflammation and severe injury,and further causes immunosuppression.However,the mechanism was not fully revealed until now.As key epigenetic modifications,5 mC and m^6A are wildly known to participate in the regulation of gene expression.The current study was to investigate the effect of IBDV infection on the bursal 5 mC and m^6A and the mRNA expressions of associated methylases and demethylases.Twenty-one-days-old commercial broilers were inoculated with IBDV,bursae were collected on day 3 and 5 post inoculation.The total bursal 5 mC and m^6A were determined using ELISA-based assays,and the mRNA expressions of the associated enzymes were detected by real-time PCR.The results showed that the bursal m^6A level and the mRNA expressions of METTL3 and METTL14 were significantly decreased on days 5 post infection;the mRNA expression of the demethylase of m^6A,FTO,was decreased on 3 and 5 days post infection(P<0.05).Bursal 5 mC level increased(P<0.05),and the mRNA level of Tet1 decreased significantly(P<0.01)at 3 days after IBDV infection;the 5 mC level at 5 days did not change,but the mRNA expressions of DNMT1 and Tet1 were remarkably decreased(P<0.01),the DNMT3 a mRNA was remarkably increased(P<0.01).The results indicated that IBDV may disturb the host immunity through dysregulating the expression of methylase and dymethylase and further modify the 5 mC and m6 A profiles.