小分子药物索非布韦对牛病毒性腹泻病毒体外复制的影响

doi:10.16431/j.cnki.1671-7236.2021.07.042

中国畜牧兽医 ›› 2021, Vol. 48 ›› Issue (7): 2653-2660.doi: 10.16431/j.cnki.1671-7236.2021.07.042

小分子药物索非布韦对牛病毒性腹泻病毒体外复制的影响

付强, 贺渊秀, 李泽宇, 塞力克·杰恩斯, 周渝新, 杨莉, 冉多良, 史慧君

新疆农业大学动物医学学院, 乌鲁木齐 830052

收稿日期:2020-12-21 出版日期:2021-07-20 发布日期:2021-07-15
通讯作者: 史慧君 E-mail:shihuijunmm@163.com
作者简介:付强(1985-),男,新疆喀什人,博士,副教授,研究方向:病原微生物致病机制,E-mail:466183013@qq.com;贺渊秀(1997-),女,新疆伊犁人,硕士生,研究方向:病原微生物致病机制,E-mail:924708744@qq.com
基金资助:
自治区天山青年计划项目青年博士科技人才培养项目（2018Q069）；自治区高校科研计划项目（XJEDU2018Y019）；国家自然科学基金项目（31760742、31902271）；新疆农业大学畜牧学博士后流动站工作（168138、168134）

Effects of Small Molecular Drug Sofosbuvir on Bovine Viral Diarrhea Virus Replication in vitro

FU Qiang, HE Yuanxiu, LI Zeyu, JIEENSI Sailike, ZHOU Yuxin, YANG Li, RAN Duoliang, SHI Huijun

College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830052, China

Received:2020-12-21 Online:2021-07-20 Published:2021-07-15

摘要/Abstract

摘要： 试验旨在研究小分子药物索非布韦是否具有抑制牛病毒性腹泻病毒（Bovine viral diarrhea virus，BVDV）复制的作用。通过MTT法测定了不同时间和不同浓度索非布韦对牛肾细胞（MDBK）增殖的影响；采用免疫荧光染色试验筛选出能有效抑制BVDV复制的索非布韦最适浓度，用实时荧光定量PCR、致细胞病变作用（CPE）和病毒半数组织细胞感染量（TCID₅₀）测定的方法检测索非布韦对BVDV复制的影响。结果显示，索非布韦处理MDBK细胞24和48 h能极显著抑制细胞增殖（P<0.01）；与对照组相比，当索非布韦浓度为800 μmol/L时免疫荧光染色检测BVDV感染MDBK细胞的双链RNA（dsRNA）含量极显著降低（P<0.01）；实时荧光定量PCR检测发现，与DMSO处理组相比，BVDV感染索非布韦处理组MDBK细胞，BVDV 5'UTR mRNA含量在病毒感染24和48 h时极显著降低（P<0.01）；BVDV感染索非布韦处理组MDBK细胞病变现象明显减弱；索非布韦处理24 h后能极显著减弱BVDV感染MDBK细胞后子代病毒颗粒的形成组与释放（P<0.01），降低病毒滴度。综合上述结果表明，小分子药物索非布韦能有效抑制BVDV体外复制。

关键词: 索非布韦; 牛病毒性腹泻病毒(BVDV); 抗病毒药物

Abstract: The aim of this study was to investigate whether Sofosbuvir,a small molecule drug,could inhibit the replication of Bovine viral diarrhea virus (BVDV).The effects of Sofosbuvir on the proliferation of bovine kidney cells (MDBK) at different time and concentrations were determined by MTT method.The optimal concentration of Sofosbuvir which could effectively inhibit BVDV replication was screened by immunofluorescence staining test.The effects of Sofosbuvir on BVDV replication were detected by Real-time PCR,CPE and TCID₅₀.The results showed that Sofosbuvir treated MDBK cells for 24 and 48 h extremely significantly inhibited the proliferation of cells (P<0.01).When Sofosbuvir was 800 μmol/L,the content of double stranded RNA (dsRNA) in MDBK cells infected with BVDV was extremely significantly lower than that in the control group (P<0.01).Real-time PCR showed that compared with DMSO treatment group,the expression of BVDV 5'UTR mRNA in Sofosbuvir treated MDBK cells was extremely significantly decreased at 24 and 48 h after infection (P<0.01).The pathological changes of MDBK cells infected with Sofosbuvir were significantly reduced.Sofosbuvir treatment for 24 h extremely significantly reduced the formation and release of virus particles in the offspring of MDBK cells infected with BVDV (P<0.01).In conclusion,Sofosbuvir could effectively inhibit BVDV replication in vitro.

Key words: Sofosbuvir; Bovine viral diarrhea virus (BVDV); anti-viral drug

中图分类号:

S852.65

付强, 贺渊秀, 李泽宇, 塞力克·杰恩斯, 周渝新, 杨莉, 冉多良, 史慧君. 小分子药物索非布韦对牛病毒性腹泻病毒体外复制的影响[J]. 中国畜牧兽医, 2021, 48(7): 2653-2660.

FU Qiang, HE Yuanxiu, LI Zeyu, JIEENSI Sailike, ZHOU Yuxin, YANG Li, RAN Duoliang, SHI Huijun. Effects of Small Molecular Drug Sofosbuvir on Bovine Viral Diarrhea Virus Replication in vitro[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2653-2660.

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小分子药物索非布韦对牛病毒性腹泻病毒体外复制的影响

Effects of Small Molecular Drug Sofosbuvir on Bovine Viral Diarrhea Virus Replication in vitro

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