大蓟水提物抗大口黑鲈虹彩病毒活性研究  

Antiviral activity of Cirsium japonicum aqueous extract (CJAE) against largemouth bass ranavirus(LMBV)

作　　者：马艳平 郝乐 冯国清 王刚 刘振兴 MA Yanping;HAO Le;FENG Guoqing;WANG Gang;LIU Zhenxing(Institute of Animal Health,Guangdong Academy of Agricultural Sciences,Key Laboratory of Livestock Disease Prevention of Guangdong Province,Guangzhou 510640,China)

机构地区：[1]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,广东广州510640

出　　处：《中国水产科学》2025年第1期103-116,共14页Journal of Fishery Sciences of China

基　　金：广东省自然科学基金项目(2022A1515012556;广东省农业科学院协同创新中心项目(XT202305));广东省农业科学院动物卫生研究所青年科技人才培育项目(PY2024008,PY2024007);广东省畜禽疫病防治研究重点实验室项目(2023B1212060040)。

摘　　要：为了探明大蓟水提物(Cirsium japonicum aqueous extract,CJAE)对大口黑鲈虹彩病毒(largemouth bass ranavirus,LMBV)的抑制活性,本研究通过体内外实验分析CJAE的抗LMBV效果,CCK-8实验证明CJAE在鳜脑细胞(Siniperca chuatsi brain cells,SCB3)上的安全浓度为≤1000μg/mL,CC50值为1487.0μg/mL;qPCR与间接免疫荧光实验表明CJAE可以有效抑制病毒核酸与MCP蛋白在SCB3细胞中的表达,EC50值为457.3µg/mL;并且CJAE在LMBV的入侵与复制阶段对病毒抑制率分别达到98.46%与99.31%。CJAE按0.5%、1.0%与1.5%剂量添加至基础饲粮,饲喂大口黑鲈15 d后,以1×106 TCID50 LMBV攻毒,结果表明CJAE可以提供35%~60%的攻毒保护率;攻毒后6 d测定大口黑鲈组织病毒拷贝数,结果表明脾脏、肝脏、肾脏与肠道的病毒载量降低87.57%~99.99%;吞噬能力测定结果表明头肾白细胞的吞噬活力上调35.88%~60.75%;mRNA表达测定结果表明CJAE激活吞噬通路基因(FcγRIa、CFL、Lyn)的表达;组织H&E染色结果表明CJAE明显减轻LMBV感染造成的大口黑鲈炎症,进一步研究发现,CJAE显著激活抗炎因子(IL-10)表达并抑制促炎因子(TNF-a、IL-1β)的表达。本研究初步揭示了CJAE抗LMBV效果与机制,为其应用于抗LMBV提供了基础。Largemouth bass ranavirus(LMBV) is widely prevalent in China,seriously threatening the safety of largemouth bass(Micropterus salmoides) aquaculture.Chinese herbal medicines have become important for screening LMBV prevention and treatment drugs.Many natural medicine plants have been shown to have direct or indirect antiviral effects.Natural medicine plants can directly impair the virus architecture,hinder the virus invasion,or block the virus entry.In multiple cases,natural medicine plant ingredients have been confirmed to inhibit the virus by stimulating the host's innate immune responses.Cirsium japonicum has many biological activities such as antivirus and anti-inflammation properties.The anti-LMBV effect of Cirsium japonicum aqueous extract(CJAE) was analyzed in vitro and in vivo to investigate the inhibitory effect on LMBV.CCK-8 tests determined that the safe concentration of CJAE on Siniperca chuatsi brain cells(SCB3) was ≤1000 μg/mL,and CC50 value was 1487.0 μg/mL.The qPCR and indirect immunofluorescence assay showed that CJAE could effectively inhibit the synthesis of viral nucleic acid and MCP protein in SCB3 cells,and the EC50 value was 457.3μg/mL.Moreover,the inhibition rates of CJAE on LMBV invasion and replication were 98.46% and 99.31%,respectively.CJAE was added to the basal diet at 0.5%,1.0%,and 1.5% to feed largemouth bass twice daily for 15days.Then a challenge test(1×106 TCID50 LMBV) was conducted.Feeding CJAE could provide 35%–60% RPS and reduced the viral load of spleen,liver,kidney and intestine of largemouth bass by 87.57%–99.99% through tissue virus copy number determination,which differed significantly from that in the control group(P<0.01).CJAE increased the phagocytic activity of head kidney leukocytes by 35.88%–60.75% through phagocytosis assay,and activated the expression of phagocytosis-related genes(FcγRIa,CFL and Lyn) through mRNA expression determination;CJAE significantly improved the inflammation of largemouth bass caused by LMBV infection through tissue H&E stainin

关 键 词：大蓟提取物 大口黑鲈虹彩病毒 EC50 吞噬 炎症 

分 类 号：S941[农业科学—水产养殖]