REV对SPF雏鸡细胞凋亡相关miRNA及其靶基因表达的影响

Influence of Apoptosis-related miRNA and Its Target Gene Experssion in SPF Chicken Infected by REV

为探究禽网状内皮组织增生病病毒(Reticuloendotheliosis virus,REV)对SPF雏鸡细胞凋亡相关信号通路的影响,通过REV感染1日龄雏鸡后,对其临床症状、大体病理学变化进行观察,并应用高通量测序及实时荧光定量PCR方法,检测并验证其法氏囊中与细胞凋亡信号通路相关的miRNA及其靶基因表达情况。结果显示:SPF雏鸡感染REV后第21天法氏囊萎缩最显著,法氏囊中gga-mir-18a-5p、gga-mir-155和gga-mir-222b-5p表达量显著下调,其靶基因分别为Caspase-6、Caspase-7和p27,上述靶基因表达量显著上调;同时还发现gga-mir-29b-3p表达量上调,其靶基因Mcl-1表达量下调。证明REV通过下调gga-mir-18a-5p、gga-mir-155和gga-mir-222b-5p以及上调gga-mir-29b-3p的表达量,来降低或增加对其靶基因的抑制,从而使促凋亡因子Caspase-6、Caspase-7和p27的表达量显著增加,同时抑凋亡因子Mcl-1表达量显著减少来促进细胞凋亡的发生。研究结果为REV致病机理的相关研究提供了理论依据。

To investigate the effect of avian reticuloendotheliosis virus （REV）on apoptosis-related signal pathway in SPF chicken, high-throughput sequencing and real-time fluorescence quantitative PCR were used to detect and verify the expression of miRNAs and their target genes related to apoptosis signal pathway in bursa of Fabricius. The results showed that the atrophy of bursa of Fabricius in SPF chicken was the most significant on the 21st day after infection with REV. The expressions of gga-mir-18a-Sp, gga-mir-155 and gga-mir-222b-Sp in bursa of Fabricius were significantly down-regulated, and the target genes were Caspase-6, Caspase-7 and p27, respectively. The expression levels of these target genes were significantly up-regulated. At the same time, it was found that the expression of gga-mir-29b-3p was up-regulated. The Mcl-1 expression was down-regulated. It was suggested that REV could decrease or increase the inhibition of its target gene by down-regulating the expression of gga-mir-18a-5pngga-mir-155 and gga-mir-222b-5p and up-regulating the expression of gga-mir-29b-3p, thus increasing the expression of Caspase-6, Caspase-7 and p27. At the same time, the expression of anti-apoptotic factor Mcl-1 was significantly reduced. REV promoted apoptosis in these ways. This study provided a theoretical basis for the study of pathogenesis of REV in the future.