摘要
【目的】探究五倍子(Galla chinensis)醇提物在石斑鱼(Epinephelus spp.)虹彩病毒(Singapore grouper iridovirus,SGIV)感染中的抗病毒效果及作用机制,为开发抗SGIV药物提供参考。【方法】通过测定细胞活力、观察细胞骨架等方法确定五倍子醇提物对石斑鱼脾脏组织细胞系(Grouper spleen cells,GS)的安全浓度,利用结晶紫染色和实时荧光定量PCR(RT-qPCR)等方法评估五倍子醇提物在细胞水平上对SGIV的抗病毒效果,进一步通过石斑鱼活体试验验证五倍子醇提物的抗SGIV效果。结合RT-qPCR和流式细胞仪的结果,分析五倍子醇提物的抗SGIV机制。【结果】当五倍子醇提物质量浓度≤12.5μg/mL时对GS细胞无显著毒性,GS细胞形态无异常。12.5、6.25、3.125μg/mL的五倍子醇提物均可降低SGIV的感染力,抗病毒效果呈现剂量依赖性。饲料中添加五倍子醇提物可使石斑鱼感染SGIV后肝脏组织内主要衣壳蛋白基因(mcp)的相对表达量显著下降(P<0.01)。五倍子醇提物可与SGIV病毒粒子相互作用,使mcp和囊膜蛋白基因(vp19)的相对表达量显著降低(P<0.01),病毒滴度显著下降(P<0.05);五倍子醇提物还可干扰SGIV感染细胞过程中的吸附、侵入和复制,药物组细胞中SGIV mcp和vp19基因的相对表达量显著降低(P<0.01)。【结论】五倍子醇提物对SGIV具有良好的体外和体内抗病毒效果,在SGIV侵染宿主细胞的吸附、侵入和复制阶段中起到干扰作用,有抗SGIV感染作用,具有成为安全、高效的抗SGIV新型渔用药物的潜力。
【Objective】To explore the antiviral effect and mechanism of methanolic extract of Galla chinensis against Singapore grouper iridovirus(SGIV)in Epinephelus spp.,and to provide reference for developing anti-SGIV drugs.【Methods】The safe concentration of the extract in grouper spleen cell lines(GS)were determined by measuring cell viability and observing cytoskeleton.The anti-SGIV effect of the extract at cellular level was evaluated by crystal violet staining and real-time quantitative PCR(RT-qPCR),the anti-SGIV effect of the extract was further verified by the in vivo tests of groupers.The anti-SGIV mechanism of the extract was analyzed together with the results of RT-qPCR and flow cytometry.【Results】When the extract mass concentration was≤12.5μg/mL,there was no discernible toxicity to GS cells,and the morphology of GS cells was normal.12.5,6.25 and 3.125μg/mL of the extract could reduce the infectivity of SGIV,and the antiviral effect was dose-dependent.The relative expression of major capsid protein gene(mcp)in liver tissue of grouper infected with SGIV was significantly decreased by dietary addition of G.chinensis methanolic extract(P<0.01).The extract could interact with SGIV virions,and relative expression levels of mcp and vesicle membrane protein gene(vp19)(P<0.01),and the viral titer decreased significantly(P<0.05).the extract also interfered with the adsorption,invasion and replication of SGIV infected cells,and the relative expression levels of mcp and vp19 genes of SGIV in the medicine group were significantly decreased(P<0.01).【Conclusion】The methanolic extract of G.chinensis has good antiviral effect on SGIV in vitro and in vivo,and interferes with the adsorption,invasion and replication of host cells infected with SGIV.It has the potential to become a safe and efficient fishery drug against SGIV.
作者
玉洁莹
黄琳
徐伟强
韦云依
王一兵
程淑
蔡佳
刘政昭
陈福彩
李鹏飞
YU Jieying;HUANG Lin;XU Weiqiang;WEI Yunyi;WANG Yibing;CHENG Shu;CAI Jia;LIU Zhengzhao;CHEN Fucai;LI Pengfei(Guangxi Academy of Sciences,Guangxi Academy of Marine Sciences,Guangxi Key Laboratory of Aquatic Biotechnology and Modern Ecological Aquaculture,Guangxi Engineering Research Center for Fishery Major Diseases Control and Efficient Healthy Breeding Industrial Technology,Nanning 530007,China;College of Food and Quality Engineering,Nanning College,Nanning 530200,China;College of Oceanography and Biotechnology,Guangxi University for Nationalities,Nanning 530007,China;Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture,College of Fisheries,Guangdong Ocean University,Zhanjiang 524088,China;Dongxing Fisheries Technology Extension Station,Dongxing 538199,China)
出处
《广东海洋大学学报》
CSCD
北大核心
2024年第6期10-18,共9页
Journal of Guangdong Ocean University
基金
国家自然科学基金(32373175)
广西自然科学基金(2024GXNSFFA010017,2022GXNSFBA035521,2022JJA130074)
国家重点研发计划(2022YFD2401200)
国家现代农业产业技术体系广西创新团队项目(nycytxgxcxtd-2021-08-02)。
