肉桂醇提物对大口黑鲈虹彩病毒的体外抗病毒作用  

Antiviral Effect of Ethanol Extracts of Cinnamomum cassia on Largemouth Bass Ranavirus

作　　者：廖文钰 竺利波[1] 刘明珠 余庆 玉洁莹 陈嘉 韦云依 黄吉利 黄琳 李鹏飞 LIAO Wenyu;ZHU Libo;LIU Mingzhu;YU Qing;YU Jieying;CHEN Jia;WEI Yunyi;HUANG Jili;HUANG Lin;LI Pengfei(Guangxi Academy of Marine Sciences,Guangxi Academy of 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 Marine Sciences,Beibu Gulf University,Qinzhou 535011,China;College of Food and Quality Engineering,Nanning College,Nanning 530200,China;Beibu Gulf Marine Industry Research Institute/China-ASEAN Modern Fishery Industry Technology Transfer Demonstration Center,Nanning 530007,China)

机构地区：[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]北部湾大学海洋学院,广西钦州535011 [3]南宁学院食品与质量工程学院,广西南宁530200 [4]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,广西南宁530007

出　　处：《广东海洋大学学报》2024年第3期17-24,共8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(2023JJG130007,2022GXNSFBA035521,2022JJA130074);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)。

摘　　要：【目的】探究肉桂醇提物在大口黑鲈(Micropterus salmoides)虹彩病毒(LMBV)感染中的抗病毒效果及作用机制,为开发抗LMBV药物提供参考。【方法】采用CCK-8试剂盒检测、结晶紫染色观察、实时荧光定量PCR(RT-qPCR)检测、病毒滴度[半数组织培养感染剂量(TCID50)]测定等方法确定肉桂醇提物在胖头鱥肌肉细胞系(FHM)上的安全工作浓度,评估肉桂醇提物在细胞水平对LMBV的抗病毒效果,分析其抗LMBV机制。【结果】肉桂醇提物在≤12.5μg/mL的质量浓度范围内对FHM细胞无明显毒性。FHM细胞与LMBV以及质量浓度分别为12.5、6.25、3.125μg/mL肉桂醇提物共孵育后,细胞病变明显减少,结晶紫染色后可见活细胞数明显升高,MCP、ICP46基因的表达量显著降低(P<0.01),且抗病毒效果有剂量依赖性。12.5μg/mL肉桂醇提物与LMBV共孵育后接入FHM细胞,细胞中MCP、ICP46基因的相对表达量以及TCID_(50)极显著下降(P<0.01);FHM细胞与经肉桂醇提物处理的LMBV共孵育后,细胞中LMBV MCP和ICP46基因的相对表达量显著低于未用肉桂醇提物处理的LMBV感染组(P<0.01);在LMBV感染FHM细胞后,接入肉桂醇提物的FHM细胞中MCP和ICP46基因的相对表达量显著下降(P<0.05);在LMBV感染FHM细胞3 h后用肉桂醇提物处理8 h的FHM细胞中MCP和ICP46基因的相对表达量显著下调(P<0.05)。【结论】肉桂醇提物可降低LMBV的感染力,干扰LMBV侵染宿主细胞过程中的吸附、侵入及复制,有良好的抗LMBV作用,可作为环保、高效的抗LMBV渔用药物候选者。【Objective】To investigate the antiviral effect and mechanism of ethanol extracts of Cinnamomum cassia in Largemouth bass virus(LMBV)infection,and to provide reference for the development of anti-LMBV drugs.【Method】The safe working concentration of cinnamon ethanol extract in fathead minnow muscle cell lines(FHM)was determined by CCK-8 kit,crystal violet staining,real-time quantitative PCR(RT-qPCR),and viral titer(TCID50),so as to evaluate the antiviral effect of cinnamon alcohol extract against LMBV at the cellular level,and to analyze the mechanism of its anti-LMBV activity.【Result】Cinnamon alcohol extract were not toxic to FHM cells at a mass concentration of≤12.5μg/mL.After co-incubation of FHM cells with LMBV and cinnamon alcohol extract at mass concentrations of 12.5,6.25,and 3.125μg/mL,respectively,the cellular lesions were significantly reduced,the number of viable cells increased significantly after crystal violet staining,and the expression of MCP and ICP46 genes was significantly reduced(P<0.01),and the antiviral effect was dose-dependent.12.5μg/mL cinnamon alcohol extract is connected to FHM cells after co-incubation with LMBV,and the relative expression of MCP and ICP46 genes and virus titer(TCID50)in the cell decreased significantly(P<0.01);after FHM cells and LMBV treated with cinnamon alcohol extract co-incubation,the relative expression of LMBV MCP and ICP46 genes in cells is significantly lower than that of the LMBV infection group not treated with cinnamon alcohol extract(P<0.01);after LMBV infects FHM cells,the relative expression of MCP and ICP46 genes in FHM cells connected to cinnamon alcohol extract decreased significantly(P<0.05);the relative expression of MCP and ICP46 genes in FHM cells treated with cinnamon alcohol extract for 8 h,3 h after LMBV infection with FHM cells,was significantly decreased(P<0.05).【Conclusion】Cinnamon alcohol extract can reduce the infectious power of LMBV,interfere with the adsorption,invasion and replication of LMBV infecting host cells,ha

关 键 词：大口黑鲈 虹彩病毒 肉桂 抗病毒作用 渔用药物 

分 类 号：S917.4[农业科学—水产科学] S941.41