Drebrin蛋白抑制剂BTP-2对猪伪狂犬病病毒增殖的影响  

Effects of Drebrin protein inhibitor BTP-2on proliferation of porcine pseudorabies virus

作　　者：王世平 王金圆 李昕蔓 唐婷 杨国宇[1] 潘佳佳[1] WANG Shiping;WANG Jinyuan;LI Xinman;TANG Ting;YANG Guoyu;PAN Jiajia(Key Laboratory for Animal Growth and Development of Henan Province,Key Laboratory for Animal Biochemistry and Nutrition,Ministry of Agriculture and Rural Affairs,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]河南农业大学农业农村部动物生化与营养重点实验室河南省动物生长发育调控重点实验室,河南郑州450002

出　　处：《中国兽医学报》2023年第9期1807-1813,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31902230);河南省重点研发与推广专项基金资助项目(222102110241)。

摘　　要：为研究肌动蛋白结合蛋白Drebrin在PRV感染过程的作用,采用化学药物BTP-2阻断Drebrin蛋白功能。首先,利用CCK-8检测BTP-2对PK-15细胞活力的影响。其次,利用荧光显微镜进行细胞形态观察,并利用流式细胞术、实时荧光定量PCR和Western blot等方法分别从细胞水平、基因水平和蛋白水平检测细胞内病毒感染量。此外,利用空斑形成单位法测定子代病毒滴度。结果显示,利用化学药物BTP-2阻断Drebrin蛋白功能,PRV子代病毒滴度显著降低,PRV-gB基因和蛋白表达量均下降,PRV-GFP细胞数量也明显降低,并呈剂量依赖性。结果表明,BTP-2抑制PRV增殖,说明Drebrin参与PRV增殖过程。该结果为进一步研究Drebrin参与PRV感染的分子机制奠定基础。Actin,the most abundant cytoskeletal protein in eukaryotic cells,is involved not only in fundamental cellular events such as cell movement and morphological maintenance,but also in the intracellular viral life cycle.Microfilaments are polymers formed by actin monomers.Actin polymerization,microfilament depolymerization,and the formation of more functional network structures are regulated by multiple actin-binding proteins.Drebrin is an actin-binding protein that induces actin filament turnover.In this study,the effect of Drebrin on PRV infection was analyzed by disrupting the function of Drebrin with BTP-2.First,the viability of PK-15cells was measured with the Cell Counting Kit-8.Then,the effect of BTP-2on PRV replication was analyzed by fluorescence microscopy,flow cytometry,real-time PCR and Western blot at cellular,gene and protein levels,respectively.In addition,the progeny virus titer was analyzed by plaque-forming unit.The results indicated that BTP-2treatment reduced the expression of the PRV-gB.It was further observed that the number of PRV-GFP positive cells in BTP-2treated cells also decreased in a dosedependent manner.The titer assay showed that BTP-2treatment restrained proliferation of the PRV-HN1201progeny virus.Taken together,these data illustrated that BTP-2inhibits PRV replication,suggesting that Drebrin participates in PRV proliferation,and provide a basis for further study of the molecular mechanism of Drebrin in PRV infection.

关 键 词：肌动蛋白 Drebrin蛋白 BTP-2 猪肾上皮细胞PK-15 猪伪狂犬病病毒 

分 类 号：S852.65[农业科学—基础兽医学]