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当前位置: 首页 > 文章 > 自噬相关蛋白Beclin-1对PHEV复制增殖的影响 中国兽医学报 2018 (5) 956-961
Position: Home > Articles > Effect of autophagy-related protein Beclin-1 on PHEV proliferation Chinese Journal of Veterinary Science 2018 (5) 956-961

自噬相关蛋白Beclin-1对PHEV复制增殖的影响

作  者：
苏晶晶;李姿;兰云刚;吕晓玲;赵魁;刘丽颖;贺文琦;高丰
单  位：
吉林大学动物医学学院;辽宁省营口市动物疫病预防控制中心
关键词：
猪血凝性脑脊髓炎病毒;Beclin-1蛋白;自噬;复制增殖
摘  要：
首先利用荧光定量PCR和Western blot的方法分别从基因水平和蛋白表达水平检测猪血凝性脑脊髓炎病毒(porcine hemagglutinating encephalomgelitis virus,PHEV)感染细胞对Beclin-1的影响,结果发现随着PHEV感染时间的增加Beclin-1表达呈现明显的上调趋势。其次,利用间接免疫荧光技术检测Beclin-1在PHEV感染后的N2a细胞中的定位变化,发现Beclin-1和PHEV呈现明显的共定位关系。随后构建Beclin-1的真核表达载体pCMV-FlagBeclin-1,将其转染至N2a细胞后接种PHEV,发现PHEV复制增殖特性并未受到显著影响。利用siRNA技术特异性敲低N2a细胞中的Beclin1水平,结果显示PHEV的表达量明显下调。本试验结果揭示了Beclin-1参与调控PHEV复制增殖的重要作用,为深入阐明PHEV致病机制提供了新的思路。
译  名：
Effect of autophagy-related protein Beclin-1 on PHEV proliferation
作  者：
SU Jing-jing;LI Zi;LAN Yun-gang;LYU Xiao-ling;ZHAO Kui;LIU Li-ying;HE Wen-qi;GAO Feng;College of Animal Science and Veterinary Medicine,Jilin University;Animal Epidemic Prevention and Control Center of Yingkou City of Liaoning Province;
关键词：
PHEV;;protein Beclin-1;;autophagy;;proliferation
摘  要：
Previous study reported that porcine hemagglutinating encephalomyelitis virus(PHEV)is able to induce autophagy.As an important autophagy-related protein,Beclin-1 is involved in regulation of some virus-induced autophagy. However, whether Beclin-1 plays a key role in PHEV infection is not clear.In order to study the effect of Beclin-1 on PHEV proliferation, the expression level of Beclin-1 was detected by quantitative PCR and western blot analysis at indicated time points post PHEV infection.Compared with the control group,Beclinl showed a significant up-regulation trend after PHEV infection.To further clarify the role of Beclin-1 in PHEV replication,immunofluorescence analysis was used to detect the localization of Beclin-1 and PHEV,and we found that Beclin-1 and PHEV were obviously colocalized in N2 a cells. Subsequently, an eukaryotic expression vector of Beclin-1 was constructed as pCMV-Flag-Beclin-1. After transfection into N2 a cells, PHEV was inoculated and found that PHEV proliferation had no significant change compared with the control group. RNA interference was used to knock down of Beclin-1 in N2 a cells and then inoculated with PHEV.The expression of PHEV was significantly down-regulated.These data revealed that Beclin-1 plays a key role in the regulation of PHEV replication and prolifera-

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