文献类型：期刊文献

中文题名：小檗碱在体外抑制神经坏死病毒增殖的作用及机制研究

英文题名：Effects and mechanisms of berberine on inhibiting nervous necrosis virus proliferation in vitro

作者：覃向谋[1];黄静[1];玉洁莹[1];余庆[1];刘明珠[1];高艳侠[1];赖俊翔[1];陈华谱[2];韩书煜[3];常彦磊[4];黄琳[1];李鹏飞[1,5]

机构：[1]广西科学院广西海洋科学院,广西水产生物技术与现代生态养殖重点实验室,广西渔业重大疫病防控与高效健康养殖产业技术工程研究中心,广西南宁530007;[2]广东海洋大学水产学院,广东省水产动物病害防控与健康养殖重点实验室,广东湛江524088;[3]广西壮族自治区水产技术推广站,广西南宁530199;[4]广州双螺旋基因技术有限公司,广东广州510700;[5]北部湾海洋产业研究院,中国-东盟现代渔业产业技术转移示范中心,农业农村部水产和畜禽养殖南宁野外科学观测研究站,广西南宁530007

年份：2026

卷号：50

期号：5

起止页码：265

中文期刊名：水产学报

外文期刊名：Journal of Fisheries of China

收录：北大核心2023、、北大核心

基金：广西自然科学基金(2023GXNSFAA026325,2022JJA130074);广西科技计划项目(桂科发[2024]102-2);来宾市科学研究与技术开发计划项目(来科产241519)。

语种：中文

中文关键词：神经坏死病毒;小檗碱;抗病毒活性;作用机制;炎症反应

外文关键词：nervous necrosis virus;berberine;antiviral activity;mechanism of action;inflammatory response

中文摘要：【目的】在细胞水平探究小檗碱(BBR)对神经坏死病毒(NNV)的抗病毒效果并阐明其作用机制。【方法】以不同浓度梯度的BBR处理卵形鲳鲹脾脏成纤维细胞(TOSF),采用光镜观察及CCK-8法确定BBR的安全工作浓度。通过细胞病变观察、实时荧光定量PCR(RT-qPCR)检测NNV衣壳蛋白(CP)基因的表达,评价BBR对NNV增殖的抑制作用。综合运用病毒预处理、时间点加药、RT-qPCR和流式细胞术等多种实验策略,系统分析BBR对病毒颗粒的直接作用及其在对NNV感染过程中吸附、侵入和复制阶段的影响。此外,通过RT-qPCR检测促炎因子IL-1β和IL-8的表达,评估BBR对NNV感染诱导炎症反应的调节作用。【结果】确定了6.25 ng/mL为BBR在TOSF细胞中的最高安全浓度。安全浓度下BBR处理TOSF细胞,能够显著减轻细胞病变效应及降低病毒CP基因的表达,抑制NNV增殖。机制研究结果显示,BBR可直接作用于NNV病毒颗粒降低其感染性,有效抑制NNV对宿主细胞的吸附与侵入,且在病毒侵入细胞后仍能显著抑制其胞内复制过程。此外,BBR还能显著下调NNV感染引起的促炎因子IL-1β和IL-8的表达。【结论】BBR能够通过直接作用病毒颗粒、阻断病毒吸附与侵入、抑制病毒在细胞内复制,并缓解宿主的炎症反应,从而发挥抗NNV效应。

外文摘要：Nervous necrosis virus(NNV)is a highly pathogenic agent that threatens global aquaculture.Berberine(BBR)is a natural alkaloid with broad-spectrum antiviral activity,yet its effect and mechanism against NNV remain unclear.This study investigated the antiviral effect and mechanism of BBR against NNV in vitro.The safe concentration of BBR was determined via CCK-8 assay.Antiviral activity was assessed through cytopathic observation and RT-qPCR detection of NNV capsid protein gene expression.Mechanistic insights were gained through virus pretreatment,time-of-addition assays,RT-qPCR,and flow cytometry,evaluating BBR’s direct virucidal effect and its interference with viral adsorption,entry,and intracellular replication.Additionally,RT-qPCR was used to examine BBR’s modulation of NNV-induced expression of pro-inflammatory cytokines IL-1βand IL-8.Results showed that BBR significantly inhibited NNV proliferation at non-cytotoxic concentrations.Mechanistically,BBR directly inactivated viral particles,blocked viral adsorption and cellular entry,and suppressed intracellular replication.It also downregulated NNV-induced overexpression of IL-1βand IL-8.In conclusion,BBR exerts anti-NNV effects through multi-target actions,including direct antiviral activity,inhibition of key infection steps,and attenuation of host inflammatory response.These findings provide a theoretical foundation for developing BBR as a green antiviral agent against NNV in aquaculture.