纳米银体外抗猪繁殖与呼吸综合征病毒的作用研究

doi:10.16431/j.cnki.1671-7236.2022.11.026

中国畜牧兽医 ›› 2022, Vol. 49 ›› Issue (11): 4356-4363.doi: 10.16431/j.cnki.1671-7236.2022.11.026

纳米银体外抗猪繁殖与呼吸综合征病毒的作用研究

楚品品, 蒋智勇, 李春玲, 勾红潮, 李艳, 蔡汝健

广东省农业科学院动物卫生研究所, 广东省畜禽疫病防治研究重点实验室, 农业农村部兽用药物与诊断技术广东科学观测实验站, 广州 510640

收稿日期:2022-04-25 出版日期:2022-11-05 发布日期:2022-11-04
通讯作者: 蔡汝健 E-mail:cairujian@gdaas.cn
作者简介:楚品品,E-mail:cpp1900@163.com。
基金资助:
广东省科技计划项目（2016B020202004）；广州市农村科技特派员项目（20212100015）；2021年英德市科技计划项目

Antiviral Effect of Silver Nanoparticles on Porcine Reproductive and Respiratory Syndrome Virus in vitro

CHU Pinpin, JIANG Zhiyong, LI Chunling, GOU Hongchao, LI Yan, CAI Rujian

Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province of Ministry of Agriculture and Rural Affairs, Key Laboratory of Livestock Disease Prevention of Guangdong Province, Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

Received:2022-04-25 Online:2022-11-05 Published:2022-11-04

摘要/Abstract

摘要： 【目的】研究纳米银（silver nanoparticles，AgNPs）体外抗猪繁殖与呼吸综合征病毒（Porcine reproductive and respiratory syndrome virus，PRRSV）的作用，并初步分析其抗病毒作用机制，为PRRSV的防控提供新思路。【方法】使用0.1875、0.375、0.75、1.5、3、6、12 μg/mL AgNPs处理Marc145细胞，采用CCK-8试剂盒评估AgNPs对Marc145细胞毒性，确定其安全浓度。通过显微观察、间接免疫荧光试验、病毒滴度测定、实时荧光定量RT-PCR方法评估AgNPs体外抗PRRSV感染Marc145细胞的效果。通过间接免疫荧光试验和实时荧光定量RT-PCR方法评估AgNPs对PRRSV的直接灭活效果。通过实时荧光定量RT-PCR方法分析AgNPs对不同感染复数（multiplicity of infection，MOI） PRRSV （0.0001~0.1）黏附和入侵Marc145细胞的影响，以及PRRSV感染Marc145细胞3、6、12、18和24 h后加入AgNPs对Marc145细胞增殖的影响。【结果】 AgNPs对Marc145细胞的最大安全浓度为1.5 μg/mL，0.375、0.75、1.5 μg/mL AgNPs均具有良好的体外抗PRRSV活性，0.375、0.75、1.5 μg/mL AgNPs均对PRRSV起一定灭活作用。AgNPs对不同MOI的毒株黏附和入侵Marc145细胞均有一定抑制作用，且不同时间（3、6、12、18、24 h）加入AgNPs对PRRSV增殖均有一定抑制作用。【结论】 AgNPs具有良好的体外抗PRRSV活性，体外抗PRRSV的作用机制包括直接灭活以及抑制病毒的黏附、入侵和增殖过程。

关键词: 纳米银(AgNPs); 猪繁殖与呼吸综合征病毒(PRRSV); 抗病毒活性; 作用机制

Abstract: 【Objective】 This study was aimed to study the antiviral effects of silver nanoparticles (AgNPs) on Porcine reproductive and respiratory syndrome virus (PRRSV) in vitro, and preliminarily analyze its mechanism, so as to provide a new idea for the prevention and control of PRRSV.【Method】 The safe concentration of AgNPs for subsequent experiments was selected from 0.1875, 0.375, 0.75, 1.5, 3, 6 and 12 μg/mL by CCK-8 kit which was used to evaluate the cytotoxicity of AgNPs to Marc145 cells.The antiviral effects of AgNPs on PRRSV were evaluated by microscopic observation, indirect immunofluorescence assay, virus titer determination and Real-time quantitative RT-PCR.Direct inactivation effect of AgNPs on PRRSV was evaluated by indirect immunofluorescence assay and Real-time quantitative RT-PCR.The effects of AgNPs on the adhesion and invasion of different multiplicity of infection (MOI) of PRRSV (0.0001 to 0.1) to Marc145 cells were analyzed by Real-time quantitative RT-PCR.The effects of AgNPs on PRRSV proliferation were analyzed by Real-time quantitative PCR after Marc145 cells were treated with AgNPs at the indicated time points that infected with PRRSV for 3, 6, 12, 18 and 24 h.【Result】 The maximum safe concentration of AgNPs to Marc145 cells was 1.5 μg/mL.There were both a certain inhibitory and inactivation effect of 0.375, 0.75 and 1.5 μg/mL AgNPs on PRRSV.The adhesion and invasion of different MOI strains to Marc145 cells was inhibited by AgNPs, and the proliferation of PRRSV was inhibited by adding AgNPs at 3, 6, 12, 18 and 24 h.【Conclusion】 AgNPs was anti-PRRSV, and the resistance was exerted through direct inactivation of PRRSV and inhibition of adhesion, invasion and proliferation of virus.

Key words: silver nanoparticles(AgNPs); Porcine reproductive and respiratory syndrome virus (PRRSV); antiviral activity; action mechanism

中图分类号:

S852.65⁺9.6

楚品品, 蒋智勇, 李春玲, 勾红潮, 李艳, 蔡汝健. 纳米银体外抗猪繁殖与呼吸综合征病毒的作用研究[J]. 中国畜牧兽医, 2022, 49(11): 4356-4363.

CHU Pinpin, JIANG Zhiyong, LI Chunling, GOU Hongchao, LI Yan, CAI Rujian. Antiviral Effect of Silver Nanoparticles on Porcine Reproductive and Respiratory Syndrome Virus in vitro[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4356-4363.

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纳米银体外抗猪繁殖与呼吸综合征病毒的作用研究

Antiviral Effect of Silver Nanoparticles on Porcine Reproductive and Respiratory Syndrome Virus in vitro

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