| 八角茴香水提物调控宿主免疫反应抗石斑鱼虹彩病毒的机制研究 |
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| 引用本文： | 李梦梦,韦红玲,刘明珠,黄帅帅,王太霞,黄琳,李鹏飞.八角茴香水提物调控宿主免疫反应抗石斑鱼虹彩病毒的机制研究[J].南方农业学报,2022,53(10):2756-2765. |
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| 作者姓名： | 李梦梦 韦红玲 刘明珠 黄帅帅 王太霞 黄琳 李鹏飞 |
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| 作者单位： | 1 河南师范大学生命科学学院, 河南新乡 453007;2 广西科学院/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, 南宁 530007;3 广西海洋天然产物与组合生物合成化学重点实验室, 广西南宁 530007;4 北部湾大学海洋学院/广西北部湾海洋生物多样性养护重点实验室, 广西钦州 535011 |
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| 基金项目： | 国家自然科学基金项目（41966004，U20A20102）；广西自然科学基金项目（2020GXNSFBA297161，AD19245022） |
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| 摘 要： | 【目的】从免疫应答角度探析八角茴香（Illicium verum Hook.f.）水提物（IVE）调控宿主免疫反应而发挥抗石斑鱼虹彩病毒（SGIV）的作用机制，为八角茴香应用于石斑鱼健康养殖产业及开发绿色、健康、高效渔用药物提供理论依据。【方法】通过光学显微镜观察及CCK-8细胞活力测定确定IVE的细胞安全工作浓度，利用实时荧光定量PCR检测IVE对宿主细胞干扰素相关基因（IFN、STAT1、PKR、ISG15、KKα、STING和IRF3）表达的影响，并以光学显微镜观察和实时荧光定量PCR分析IVE在细胞水平抗SGIV的效果，通过石斑鱼活体试验进一步验证IVE的抗SGIV效果。【结果】IVE细胞水平的安全工作浓度≤0.50 mg/mL； IVE预处理GS细胞2 h可显著（P<0.05，下同）或极显著（P<0.01，下同）激活干扰素相关基因（IFN、STAT1、PKR、IKKα、IRF3和STING）的表达。SGIV感染GS细胞24和48 h时，经IVE预处理的细胞病变效应（CPEs）明显少于未使用IVE预处理的GS细胞，且细胞内SGIV的MCP基因相对表达量极显著降低，表明IVE可有效抑制SGIV感染；此外，IVE可增强SGIV感染GS细胞中干扰素相关基因的表达，具体表现为IFN、STAT1和IRF3基因在病毒感染后24 h极显著上调，PKR、IKKα、ISG15和STING基因在病毒感染后24和48 h呈显著或极显著上调趋势。在石斑鱼活体试验中，IVE与SGIV混合腹腔注射后石斑鱼脾脏中SGIV的MCP基因相对表达量在病毒感染后24和48 h均极显著低于仅注射SGIV的石斑鱼。【结论】IVE具有抑制SGIV感染的功能，其作用机制可能是通过诱导干扰素相关基因表达而激活宿主的抗病毒状态，从而发挥间接抗SGIV效果；也说明八角茴香在水产疫病防控中具有研发成渔用抗病功能制剂的潜力。
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| 关 键 词： | 石斑鱼虹彩病毒（SGIV） 八角茴香水提物（IVE） 免疫反应 抗病毒机制 干扰素 |
| 收稿时间： | 2021-12-30 |
Mechanism of Illicium verum Hook. f. extracts to regulate host immune response against Singapore grouper iridovirus |
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| LI Meng-meng,WEI Hong-ling,LIU Ming-zhu,HUANG Shuai-shuai,WANG Tai-xia,HUANG Lin,LI Peng-fei.Mechanism of Illicium verum Hook. f. extracts to regulate host immune response against Singapore grouper iridovirus[J].Journal of Southern Agriculture,2022,53(10):2756-2765. |
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| Authors: | LI Meng-meng WEI Hong-ling LIU Ming-zhu HUANG Shuai-shuai WANG Tai-xia HUANG Lin LI Peng-fei |
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| Abstract: | 【Objective】 To explore mechanism of Illicium verum Hook.f. extracts(IVE) to regulate host immune response against Singapore grouper iridovirus(SGIV) from the perspective of immune response, so as to provide a theoreti-cal basis for utilizing I. verum Hook. f. to be developed into a green, safe and efficient fishery drug for grouper breeding industry.【Method】 The safe working concentration of IVE was determined by light microscope observation and CCK-8 cell viability assay;the real-time fluorescence quantitative PCR(qRT-PCR) method was used to analyze the effect of IVE on the host interferon(IFN) related genes immune factors(IFN, STAT1, PKR, ISG15, IKKα, STING and IRF3) expression;then the effect of IVE against SGIV at the cellular level was analyzed by light microscopy and qRT-PCR methods. The anti-SGIV effect of IVE was further verified through in vivo grouper experiment.【Result】 The safe working concentration of IVE was ≤ 0.5 mg/mL;GS cells IVE-pretreated for 2 h could significantly(P<0.05, the same below) or extremely significantly(P<0.01, the same below) activate the expression of genes involved in STAT1 system(STAT1, PKR, IKKα, IRF3 and STING). At 24 h and 48 h of SGIV infection, the cytopathic effects(CPEs) of IVE-pretreated cells occurred less significantly than GS cells without IVE pretreatment;and MCP gene relative expression of SGIV decreased extremely significantly, suggesting that IVE could effectively inhibit the infection of SGIV. Moreover, IVE could enhance IFN-related gene expression of SGIV-infected GS cells as IFN, STAT1 and IRF3 genes were extremely significantly up-regulated 24 h after infection and PKR, IKKα, ISG15 and STING genes were significantly or extremely significantly up-regulated 24 h and 48 h after infection. In the in vivo experiment, the relative expression of the MCP gene of SGIV in the spleen of the grouper injected with IVE and SGIV was extremely significantly lower than that of the grouper injected with SGIV at 24 and 48 h.【Conclusion】 IVE can inhibit SGIV infection probably through the mechanism which induces expression of IFNrelated genes to activate anti-viral function of host, thus indirectly fight against SGIV. It also suggests that I. verum Hook. f. has the potential to be developed into an anti-aquatic virus drug. |
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