【作者】 吕晓玲； 贺文琦； 高丰； 赵魁； 李姿； 胡诗雨；

【Author】 LV Xiao-ling;HE Wen-qi;GAO Feng;ZHAO Kui;LI Zi;HU Shi-yu;College of Veterianry Medicine,Jilin University;

【机构】 吉林大学动物医学学院；

【摘要】 为研究PHEV感染过程中宿主mi R-21a-5p对PHEV复制的影响,本研究利用RT-PCR方法对PHEV感染过程中细胞内mi R-21a-5p的表达变化进行检测,发现mi R-21a-5p的表达量明显升高。之后,将mi R-21a-5p模拟体或抑制剂转染N2a细胞,接种PHEV后利用RT-PCR、Western blotting方法检测PHEV相对表达量,结果显示抑制mi R-21a-5p的表达会使PHEV复制增殖显著下降,反之使PHEV表达量升高。其次,通过生物信息学软件预测mi R-21a-5p的靶基因并进行功能分析,发现参与调控突触传递、轴突导向的支架蛋白Caskin1可能为其靶基因。双荧光素酶报告系统、RT-PCR、Western blotting结果均证实Caskin1为mi R-21a-5p的靶基因,并且当抑制细胞内源性Caskin1表达后,PHEV复制量明显升高。综上所述,在PHEV致病过程中mi R-21a-5p可能通过靶向Caskin1来调控病毒复制。研究结果不仅为mi RNA参与调控PHEV复制提供数据支持,也为深入研究PHEV的致病机制提供新的思路。更多还原

【Abstract】 The aim of this study was to investigate the effect of mi R-21 a-5 p on PHEV replication during PHEV infection In.The expression of mi R-21 a-5 p in PHEV-infected cells was examined by RT-PCR.The results showed that the expression of mi R-21 a-5 p was significantly increased in PHEV-infected N2 a cells.Next,the expression of PHEV was detected by RT-PCR,Western blotting after mi R-21 a-5 p mimic or the inhibitor was transfected into N2 a cells.The results showed that inhibitionof mi R-21 a-5 p expression resulted in a significant decrease in PHEV replication and proliferation.Furthermore,the target gene of mi R-21 a-5 p was predicted by bioinformatics software.We found that Caskin1 which involved in the regulation of synaptic transmission and axon-directed scaffold minght be the target gene.Double-luciferase reporter system,RT-PCR and Western blotting showed that Caskin1 was the target gene of mi R-21 a-5 p.And the expression of PHEV was significantly increased when the expression of endogenous Caskin1 was inhibited.In summary,mi R-21 a-5 p might be regulate viral replication during PHEV pathogenesisused by targeting Caskin1.These findings may represent new clues for mi RNA involved in the regulation of PHEV replication,and help the development of strategies to the further study of the pathogenesis of PHEV.更多还原