PDF(Pubmed)
Abstract：
Infectious hematopoietic necrosis virus (IHNV) is an important pathogen that causes significant economic losses to salmon trout farming. Although vaccines have been invented for the treatment of IHNV, findings from our previous survey show that breeding enterprises and farmers require effective oral drugs or immune enhancers. However, studies on the development of oral drugs are limited. In the present study, we used bioinformatics methods to predict the protein targets of andrographolide (Andro) in IHNV. Cells were infected with IHNV, and the effect of andrographolide was explored by evaluating the expression levels of genes implicated in oxidative stress, activities of antioxidant enzymes, and the expression of genes implicated in apoptosis and necrosis. In the present study, cells were divided into NC, IHNV, IHNV+10 μM andrographolide, and IHNV+20 μM andrographolide groups. qRT-PCR was performed to determine the expression level of genes, and an antioxidant enzyme detection kit was used to evaluate the activities of antioxidant enzymes. Fluorescent staining was performed using a reactive oxygen species detection kit (ROS) and Hoechst 33342/PI double staining kit, and the mechanism of alleviation of apoptosis and oxidative stress andrographolide after IHNV infection was determined. The results indicated that andrographolide inhibits viral growth by binding to the NV protein of IHNV and increasing the antioxidant capacity of the body through the CTSK/BCL2/Cytc axis, thereby inhibiting the occurrence of IHNV-induced apoptosis. This is the first study to explore the antagonistic mechanism of action of andrographolide in alleviating IHNV infection. The results provide valuable information on alternative strategies for the treatment of IHNV infection during salmon family and provide a reference for the use of andrographolide as an antioxidant agent in agricultural settings.
摘要：
传染性造血器官坏死病毒（IHNV）是一种重要的病原体，对鲑鱼鳟鱼养殖造成重大的经济损失。尽管已经发明了用于治疗IHNV的疫苗，我们先前的调查结果表明，育种企业和农民需要有效的口服药物或免疫增强剂。然而,关于口服药物开发的研究是有限的。在本研究中,我们使用生物信息学方法来预测IHNV中穿心莲内酯（Andro）的蛋白质靶标。用IHNV感染细胞,并通过评估与氧化应激有关的基因的表达水平来探索穿心莲内酯的作用，抗氧化酶的活性，以及与细胞凋亡和坏死有关的基因的表达。在本研究中,细胞被分成NC，IHNV,IHNV+10μM穿心莲内酯，和IHNV+20μM穿心莲内酯组。进行qRT-PCR以确定基因的表达水平，并使用抗氧化酶检测试剂盒评估抗氧化酶的活性。使用活性氧检测试剂盒（ROS）和Hoechst33342/PI双重染色试剂盒进行荧光染色，确定了穿心莲内酯减轻IHNV感染后细胞凋亡和氧化应激的机制。结果表明，穿心莲内酯通过与IHNV的NV蛋白结合，并通过CTSK/BCL2/Cytc轴增加机体的抗氧化能力，从而抑制病毒的生长，从而抑制IHNV诱导的细胞凋亡的发生。这是首次探讨穿心莲内酯缓解IHNV感染的拮抗作用机制。该结果为鲑鱼家族中IHNV感染的替代治疗策略提供了有价值的信息，并为在农业环境中使用穿心莲内酯作为抗氧化剂提供了参考。
