塞内卡病毒A结构蛋白VP2诱导PK-15细胞自噬的研究  

Autophagy in PK-15 Cells Induced by the Structural Protein of Senecavirus A VP2

作　　者：王晶[1] 郭禹 赵云环 顾文源[1,3] 刘涛[1,4] 翟刚 张帅[1] 王丙雷 左玉柱[1,2] 范京惠 WANG Jing;GUO Yu;ZHAO Yunhuan;GU Wenyuan;LIU Tao;ZHAI Gang;ZHANG Shuai;WANG Binglei;ZUO Yuzhu;FAN Jinghui(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China;Hebei Veterinary Biotechnology Innovation Center,Hebei Agricultural University,Baoding 071001,China;Animal Diseases Control Center of Hebei,Shijiazhuang,050053,China;Ringpu(Baoding)Biological Pharmaceutical Co.,Ltd.,Baoding 071000,China)

机构地区：[1]河北农业大学动物医学院,保定071001 [2]河北省兽医生物技术创新中心,保定071001 [3]河北省动物疫病预防控制中心,石家庄050053 [4]瑞普(保定)生物药业有限公司,保定071000

出　　处：《病毒学报》2022年第2期431-438,共8页Chinese Journal of Virology

基　　金：河北省重点研发计划项目(项目号:19226622D),题目:猪重要新发疫病病原和抗体关键检测技术研究;河北省农业产业技术体系生猪创新团队(项目号:HBCT2018110207),题目:河北省农业产业技术体系生猪创新团队疫病防控岗。

摘　　要：本研究在塞内卡病毒A(Senecavirus A,SVA)诱导自噬的基础上,着重探究VP2蛋白在细胞自噬过程中的作用。构建VP2基因真核表达载体pcDNA3.1-VP2,将其转染至PK-15细胞,通过检测自噬蛋白和相关基因的表达情况,明确VP2蛋白对细胞自噬的影响。结果显示,本研究成功构建了pcDNA3.1-VP2真核表达载体,且SVA VP2基因在PK-15细胞中正常表达;与对照组相比,VP2蛋白显著上调LC3蛋白的表达水平(P<0.01);同时,自噬基因LC3、Beclin-1和ATG5转录水平均显著提高(P<0.01)。综上所述,本研究证实SVA VP2蛋白可诱导PK-15细胞自噬,且VP2蛋白与自噬蛋白和基因表达水平呈正相关,为进一步研究病毒感染与致病机制打下基础。Based on the induction of autophagy by Senecavirus A(SVA),we explored the role of VP2 protein in autophagy. A eukaryotic expression vector of VP2,pcDNA3.1-VP2 was constructed,and transfected into PK-15 cells. The effect of VP2 protein on autophagy was evaluated by measuring expression of autophagy-related proteins and related genes. Results showed that the pcDNA3.1-VP2 eukaryotic expression vector was constructed,and VP2 gene of SVA was expressed in PK-15 cells. Compared with the control group,VP2 protein increased expression of LC3 protein significantly(P<0.01). Simultaneously,transcription of the autophagy-related genes LC3,Beclin-1 and ATG5 was increased significantly(P<0.01). In summary,we demonstrated that the VP2 protein of SVA could induce autophagy in PK-15 cells,and the expression of VP2 protein was positively correlated with expression of autophagy-related proteins and genes. These data lay the foundation for further research on SVA infection and pathogenic mechanisms.

关 键 词：塞内卡病毒A(SVA) 结构蛋白VP2 PK-15细胞 自噬 

分 类 号：S852.65[农业科学—基础兽医学]