刺五加水提物抗鳜蛙虹彩病毒活性及其对大口黑鲈免疫调节作用的研究

doi:10.16431/j.cnki.1671-7236.2023.03.035

中国畜牧兽医 ›› 2023, Vol. 50 ›› Issue (3): 1207-1217.doi: 10.16431/j.cnki.1671-7236.2023.03.035

刺五加水提物抗鳜蛙虹彩病毒活性及其对大口黑鲈免疫调节作用的研究

马艳平¹, 徐晶晶², 杨宏伟¹, 郝乐¹, 贾梦岩³, 陶子骞², 王敬雅⁴, 黄文⁵, 彭凯⁵, 刘振兴¹

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, 农业农村部兽用药物与诊断技术广东科学观测实验站, 广东省农业科学院协同创新中心, 广州 510640;
2. 仲恺农业工程学院动物科技学院, 广州 510225;
3. 广东海洋大学水产学院, 湛江 524088;
4. 华南农业大学兽医学院, 广州 510642;
5. 广东省农业科学院动物科学研究所, 广东省畜禽育种与营养研究重点实验室, 农业农村部华南动物营养与饲料重点实验室, 广州 510640

收稿日期:2022-09-20 出版日期:2023-03-05 发布日期:2023-03-02
通讯作者: 刘振兴 E-mail:liuzhenxing@gdaas.cn
作者简介:马艳平,E-mail:mayanping2292@163.com;徐晶晶,E-mail:2434399243@qq.com。
基金资助:
广东省农业科学院协同创新中心项目(XT202232、XT202305);佛山市财政专项资金-2022年度共建广东农业科技示范市项目(抗大口黑鲈虹彩病毒中草药饲料的研发)

Anti-MRV Activity of Acanthopanax senticosus Aqueous Extract (ASAE) and Its Immunomodulatory Effects on Micropterus salmoides

MA Yanping¹, XU Jingjing², YANG Hongwei¹, HAO Le¹, JIA Mengyan³, TAO Ziqian², WANG Jingya⁴, HUANG Wen⁵, PENG Kai⁵, LIU Zhenxing¹

1. Collaborative Innovation Center of GDAAS, Scientific Observation and Experiment Station of Veterinary Drugs and Diagnostic Techniques of Guangdong Province, 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;
2. College of Animal Science and Technology, Zhongkai University of Agriculture and Engineering, Guangzhou 510225, China;
3. Fisheries College, Guangdong Ocean University, Zhanjiang 524088, China;
4. College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China;
5. Key Laboratory of Animal Nutrition and Feed Science in South China, Ministry of Agriculture and Rural Affairs, Guangdong Key Laboratory of Animal Breeding and Nutrition, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

Received:2022-09-20 Online:2023-03-05 Published:2023-03-02

摘要/Abstract

摘要： 【目的】研究刺五加水提物(Acanthopanax senticosus aqueous extract,ASAE)对鳜蛙虹彩病毒(Mandarin ranavirus,MRV)的抑制作用及其对大口黑鲈免疫调节功能的影响。【方法】用不同剂量ASAE(1.2、1.0、0.8、0.6、0.4和0.2 mg/mL)与MRV同时作用鳜鱼脑细胞系(SCB3),检测ASAE对MRV的抑制活性。在基础饲粮中添加5‰ ASAE,连续饲喂大口黑鲈15 d,实时荧光定量PCR和流式细胞术分析ASAE对大口黑鲈免疫功能的影响,并以1×10⁷ TCID₅₀/mL MRV腹腔注射攻毒,分析ASAE抗MRV保护效果。【结果】1.2、1.0、0.8、0.6、0.4和0.2 mg/mL ASAE对MRV抑制率分别为95.27%、91.36%、84.55%、60.05%、32.16%和2.49%。实时荧光定量PCR结果显示,ASAE诱导大口黑鲈白介素-10(interleukin-10,IL-10)、酪氨酸激酶(tyrosine-protein kinase,Lyn)、肌动蛋白相关蛋白(actin related protein 2/3 complex,subunit 5,ARPC5)、Fcγ受体Ⅰa(Fcγ receptor Ⅰa,FcγRⅠa)等Fcγ受体吞噬通路相关基因显著性上调表达(P＜0.05);流式细胞术分析结果显示,添加ASAE明显提高了大口黑鲈头肾白细胞吞噬能力(吞噬率80.9%)。攻毒试验结果显示,ASAE对MRV的相对保护率为30.48%。与对照组相比,ASAE明显延缓了大口黑鲈死亡时间,且添加ASAE组脑、脾脏、中肠、后肠组织的病毒拷贝数显著降低(P＜0.05)。【结论】ASAE可显著激活大口黑鲈吞噬活性,发挥免疫调节作用,体内外试验均表明ASAE具有优异的抗MRV效果,可以有效预防MRV感染。

关键词: 刺五加水提物(ASAE); 鳜蛙虹彩病毒(MRV); 大口黑鲈; 体外试验; 体内试验; 吞噬影响

Abstract: 【Objective】 The aim of this study was to investigate the inhibitory effect of Acanthopanax senticosus aqueous extract (ASAE) on Mandarin ranavirus (MRV) and the immunomodulatory effects of Micropterus salmoides.【Method】 In this study,the inhibitory activity of ASAE on MRV in Siniperca chuatsi brain cell 3 lines (SCB3) was detected with different doses of ASAE (1.2,1.0,0.8,0.6,0.4 and 0.2 mg/mL) and MRV. 5‰ ASAE was added to the basal diet and fed Micropterus salmoides for 15 days,the phagocytosis effect of ASAE on Micropterus salmoides was analyzed by quantitative Real-time PCR and flow cytometry,and the protective effect of ASAE on MRV was determined by MRV infection test (1×10⁷ TCID₅₀/mL MRV,intraperitoneal injection).【Result】 The inhibition test results showed that MRV inhibition rates were 95.27%,91.36%,84.55%,60.05%,32.16% and 2.49% when the different concentrations of ASAE (1.2,1,0.8,0.6,0.4 and 0.2 mg/mL) were added into SCB3 cell lines.The quantitative Real-time PCR results showed that the ASAE induced significant up-regulation of cell phagocytosis pathway genes including interleukin-10 (IL-10),tyrosine-protein kinase (Lyn) and actin related protein 2/3 complex,subunit 5 (ARPC5), and Fcγ receptor Ⅰa (FcγR Ⅰa) in Micropterus salmoides (P＜0.05).The flow cytometry analysis showed that the ASAE significantly increased the phagocytosis ability of head kidney leukocytes of Micropterus salmoides (80.9% phagocytosis rate).The infection results showed that the relative protective rate of ASAE group was 30.48%.Compared with the control group,ASAE significantly delayed the process of infection and death of Micropterus salmoides. The virus copy numbers in brain,spleen,midgut and hindgut tissue of ASAE group were significantly decreased (P＜0.05).【Conclusion】 The results proved that ASAE significantly activated the phagocytic activity of Micropterus salmoides and played an excellent immunoregulatory role.Through the vitro and vivo tests,ASAE had excellent anti-MRV effect and could effectively prevent MRV infection.

Key words: Acanthopanax senticosus aqueous extract (ASAE); Mandarin ranavirus (MRV); Micropterus salmoides; in vitro test; in vivo test; phagocytic effect

中图分类号:

S853.7

马艳平, 徐晶晶, 杨宏伟, 郝乐, 贾梦岩, 陶子骞, 王敬雅, 黄文, 彭凯, 刘振兴. 刺五加水提物抗鳜蛙虹彩病毒活性及其对大口黑鲈免疫调节作用的研究[J]. 中国畜牧兽医, 2023, 50(3): 1207-1217.

MA Yanping, XU Jingjing, YANG Hongwei, HAO Le, JIA Mengyan, TAO Ziqian, WANG Jingya, HUANG Wen, PENG Kai, LIU Zhenxing. Anti-MRV Activity of Acanthopanax senticosus Aqueous Extract (ASAE) and Its Immunomodulatory Effects on Micropterus salmoides[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1207-1217.

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刺五加水提物抗鳜蛙虹彩病毒活性及其对大口黑鲈免疫调节作用的研究

Anti-MRV Activity of Acanthopanax senticosus Aqueous Extract (ASAE) and Its Immunomodulatory Effects on Micropterus salmoides

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