败酱草水提物对石斑鱼虹彩病毒的抗病毒作用

Antiviral Effects of Thlaspi arvense Linn.Water Extracts Against Grouper Iridovirus Infection

关键词:  败酱草水提物  石斑鱼虹彩病毒  抗病毒  细胞活力  核酸适配体  渔用药物

Key words:  The water extracts of Thlaspi arvense Linn.  grouper iridovirus  antiviral  cell viability  aptamer  fishery drugs

摘要:

石斑鱼是名贵的海水养殖鱼类，随着高密度、集约化养殖的发展，各类水产疫病频繁暴发。石斑鱼虹彩病毒（Grouper iridovirus）是石斑鱼养殖中最严重的病毒性病原之一，会导致鱼大量死亡，因此研究可抑制此病毒感染的药物对石斑鱼养殖业具有重大意义。败酱草为我国传统中药，现代药理学证明，败酱草及其活性成分具有抗病毒、抗菌、抗肿瘤等多种生物活性。本研究首先采用光学显微镜观察、细胞活力测定实验确定败酱草水提物的细胞安全浓度；然后结合实时荧光定量PCR技术（RT-qPCR）和基于核酸适配体Q5的荧光分子探针检测技术（Aptamer Q5-based fluorescent molecular probe assay，Q5-AFMP）等方法分析败酱草水提物在细胞水平对石斑鱼虹彩病毒的抗病毒效果。其结果显示，低于2.50 mg/mL的败酱草水提物对细胞无明显的毒性作用，即败酱草水提物对石斑鱼细胞的安全浓度为≤ 2.50 mg/mL，高于2.50 mg/mL的败酱草水提物会降低细胞活力，并对细胞有显著的毒性作用。光学显微镜观察、细胞活力测定、RT-qPCR和Q5-AFMP技术的检测结果均证明败酱草水提物在体外对石斑鱼虹彩病毒感染具有良好的抑制作用。说明细胞安全浓度的败酱草水提物具有良好的抗石斑鱼虹彩病毒作用，具有开发为高效的抗病毒渔用药物的潜力。

Abstract:

Grouper is a precious aquaculture fish. With the development of high-density and intensive aquaculture,various kinds of aquatic epidemics break out frequently. The grouper iridovirus is one of the most serious pathogens in grouper culture,which can cause a large number of fish deaths in a very short time. Therefore,it is of great significances to develop antiviral drugs that inhibit this virus infection for grouper aquaculture industry.Thlaspi arvense Linn.is a traditional Chinese medicine,and modern pharmacology has proved that Thlaspi arvense Linn.and its active ingredients have various biological activities such as antiviral,antibacterial and antitumor. In this study,firstly the optical microscope observation and cell viability assay were used to determine the cellular safe concentration of the water extracts of Thlaspi arvense Linn. Then RT-qPCR and aptamer Q5-based fluorescent molecular probe assay (Q5-AFMP) and other methods were used to analyze the antiviral effect of water extracts of Thlaspi arvense Linn.on the grouper iridovirus at the cellular level. The results showed that the waters extracts of Thlaspi arvense Linn.below 2.50 mg/mL had no obvious toxic effect on the cells,that was,the safe concentration of water extracts of Thlaspi arvense Linn.on grouper cells was ≤ 2.50 mg/mL. Concentration of water extracts of Thlaspi arvense Linn.higher than 2.50 mg/mL could reduce cell viability and had significant toxic effects on cells. The results of light microscope observation,cell viability assay,RT-qPCR and Q5-AFMP all proved that the water extracts of Thlaspi arvense Linn.had good inhibitory effects on grouper iridovirus infection in vitro. Overall,the results showed that the water extracts of Thlaspi arvense Linn.with appropriate concentration had excellent antiviral effects and had the potential to develop an efficient antiviral fishery drugs.