JEV入侵BHK-21细胞内吞途径的初步研究  

Preliminary research on the endocytic pathway of JEV entry into BHK-21 cells

作　　者：曹三杰[1] 黄恒[1] 刘凯[1] 石双艳 黄小波[1,2,3] 文心田[1,2,3] 

机构地区：[1]四川农业大学预防兽医研究所猪病研究中心,雅安625014 [2]四川农业大学动物医学院人兽共患病研究室,成都611130 [3]农业部兽用药物与兽医生物技术四川科学观测实验站,雅安625014

出　　处：《中国人兽共患病学报》2013年第8期766-770,共5页Chinese Journal of Zoonoses

基　　金：公益性行业(农业)科研专项经费项目(No.201203082)资助~~

摘　　要：目的利用Real-time RT-PCR技术研究,对乙型脑炎病毒入侵细胞的内吞途径进行初步研究。方法选用氯丙嗪、制霉菌素和细胞松弛素D3种药物分别处理细胞,以分别阻滞网格蛋白介导内吞途径、细胞膜穴样内陷途径和巨胞饮途径,然后进行JEV感染实验,通过Real-time RT-PCR方法测定感染后1h、12h、24h细胞内的病毒拷贝数。结果与仅实施感染实验、未进行药物处理的对照组比较,3种药物处理组的细胞内病毒拷贝数在感染后各时相点均呈现不同程度的下降,在感染1h、12h和24h后,氯丙嗪处理组细胞内病毒拷贝数分别下降了59.58%,53.34%和61.35%;细胞松弛素D组细胞内病毒拷贝数分别下降了23.59%,30.77%和30.14%;制霉菌素组细胞内病毒拷贝数分别下降了35.56%,34.40%和34.40%。结论乙型脑炎病毒主要利用网格蛋白介导内吞途径进入BHK-21细胞。We applied real-time PCR to study the endocytic pathway of JEV entry into BHK-21 cells. BHK-21 ceils were treated with Chlorpromazine, Nystatin and Cytochalasin D to block clathrin-mediated endocytosis, caveolae-mediated endocyto sis, and phagocytosis, respectively. Then those treated cells were infected with JEV. Real-time RT-PCR assay was used in this study to estimate the intracellular virus copy-number at 1 hour, 12 hours, and 24 hours after infection, respectively. Compared with control group, the intracellular virus copy numbers of three groups treated with drug were in different degrees of decline. The change showed in the group treated with Chlorpromazine was the most obvious, and the intracellular virus copy number decreased by 59.58%, 70.20% and 61.35%, respectively; the intracellular virus copy number in the cytochalasin D group de creased by 23.59%, 55.79% and 30.14%, and in the nystatin group decreased by 35.56%, 58.10% and 34.40%, respective ly. Results indicate that JEV infect BHK-21 cells by clathrin-mediated endocytosis.

关 键 词：乙型脑炎病毒 实时荧光定量RT—PCR 内吞途径 

分 类 号：R373[医药卫生—病原生物学]