miR-21通过PI3K/Akt通路抑制猪瘟病毒复制的机制的研究  

miR-21 inhibition of classical swine fever virus replication through the PI3K/Akt signaling pathway

作　　者：董小英[1,2] 彭国良[1,2] 许崇波[2] 胡鸿惠[2] 南文金[2] 吴静波[2] 黄健强[2] 唐胜球[1,2] 

机构地区：[1]韶关学院英东农业科学与工程学院,广东韶关512005 [2]韶关学院粤北生猪生产及疾病防控协同创新发展中心,广东韶关512005

出　　处：《中国预防兽医学报》2016年第8期595-599,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然基金(31372394);广东省自然基金(2016A030307043;2015A030310094;2014A030313699)

摘　　要：为探索miR-21调控猪瘟病毒(CSFV)复制的分子机制,本实验采用猪瘟病毒感染体外培养的PK-15细胞,采用RT-PCR、western blot等方法检测miR-21表达、猪瘟病毒复制、PI3K/Akt通路变化及其三者之间的联系。结果表明,猪瘟病毒感染导致细胞miR-21表达水平下调,而PI3K/Akt通路相关因子磷酸化Akt蛋白表达显著上调。将miR-21类似物转染细胞后,猪瘟病毒滴度和RNA水平明显降低,而且PI3K/Akt通路受到阻遏。然而,采用阻断剂LY294002阻断细胞PI3K/Akt通路,结果显示,猪瘟病毒复制有所减弱,但作用不显著,而miR-21抑制猪瘟病毒复制的作用明显减弱。以上结果表明,miR-21可以通过PI3K/Akt通路抑制猪瘟病毒复制。To study the signaling pathway of the microRNA-21 （miR-21） impact on replication of classical swine fever virus （CSFV） in vitro, the miR-21 expression, CSFV replication, the activity of the PI3K/Akt signaling pathway and the the relationships among these factors were detected and analyzed in CSFV infected PK-15 cells. The results confirmed that CSFV infection reduced miR-21 expression, but significantly enhanced the protein expression of the PI3K/Akt pathway related factor of phosphorylated Akt. In addition, following the transfection of miR-21 analogs, the PK-15 cells were infected with CSFV, as the results, the viral RNA levels and viral titers were significantly decreased, and the PI3K/Akt signaling pathway was suppressed. To fiLrther determine the target of miR-21 in CSFV replication, cell PI3K./Akt signaling pathway was blocked using inhibitor LY294002. It was indicated that the block of PI3K/Akt signaling pathway reduced CSFV replication, but the effect was not significant. However, inhibition of CSFV replication induced by miR-21 was significantly weakened following the block of this signaling pathway. In conclusion, it was suggested that miR-21 might inhibit CSFV replication through suppression of the PI3K/Akt signaling pathway.

关 键 词：MIR-21 猪瘟病毒 复制 PI3K/AKT 

分 类 号：S852.65[农业科学—基础兽医学]