State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
College of Advanced Agricultural Sciences, University of Chinese Academy of Sciences, Beijing 100049, China.
Int J Mol Sci. 2024 Nov 4;25(21):11852. doi: 10.3390/ijms252111852.
Hemorrhagic disease caused by grass carp reovirus (GCRV) infection is a major problem affecting the grass carp aquaculture industry. Therefore, inhibiting the spread of GCRV infection is of great economic significance. Herein, we sequenced five tissues (gill, liver, intestine, kidney, and muscle) from grass carp before and after GCRV infection using data-independent acquisition proteomic and untargeted metabolomic technologies, and quantitatively identified 10,808 proteins and 4040 metabolites. Then, we analyzed the differentially expressed proteins (DEPs) and metabolites (DEMs) before and after GCRV infection in the five tissues. Gene ontology analysis revealed that the five tissue DEPs were enriched in metabolic, including carbohydrate and lipid metabolic processes. Chemical taxonomy analysis showed that the categories of DEMs mainly included carbohydrates and lipids, such as fatty acids, glycerophospholipids, steroids, and their derivatives. Both the proteomic and the metabolomic data showed that GCRV affected the carbohydrate and lipid metabolism in the host. Shared pathway analysis was performed at both the protein and metabolic levels, showing significant enrichment of the glycolysis and pentose phosphate pathways ( < 0.001). Further analysis of glycolysis and pentose phosphate pathway inhibitors revealed that these two pathways are important for GCRV replication. As the kidney was the most affected among the five tissues, we analyzed the butanoate metabolism in the kidney, which revealed that most of the differentially expressed proteins and differently expressed metabolites in the butanoate metabolism were related to the TCA cycle. Further investigation showed that fumaric acid, an intermediate product in the TCA cycle, significantly inhibited GCRV replication in the CIK cells ( < 0.001), and that this inhibitory effect may be related to its induction of interferon system activation. The addition of fumaric acid to feed increased the survival rate of juvenile grass carp by 19.60% during GCRV infection, and protected the tissues of those infected with GCRV, making it a potential anti-GCRV feed additive. Our results provide new perspectives on GCRV pathogenesis and antiviral strategies for grass carp.
草鱼出血病是一种由草鱼呼肠孤病毒（GCRV）感染引起的疾病，是严重影响草鱼养殖业的主要问题。因此，抑制 GCRV 感染的传播具有重要的经济意义。本研究采用非靶向代谢组学和数据非依赖性采集蛋白质组学技术，对草鱼感染 GCRV 前后的 5 种组织（鳃、肝、肠、肾和肌肉）进行测序，定量鉴定了 10808 种蛋白质和 4040 种代谢物。然后，我们分析了 5 种组织中 GCRV 感染前后的差异表达蛋白（DEPs）和代谢物（DEMs）。GO 分析显示，5 种组织的 DEPs 主要富集在代谢过程中，包括碳水化合物和脂质代谢过程。化学分类分析表明，DEMs 的主要类别包括碳水化合物和脂质，如脂肪酸、甘油磷脂、类固醇及其衍生物。蛋白质组学和代谢组学数据均显示，GCRV 影响宿主的碳水化合物和脂质代谢。在蛋白质和代谢水平上进行的共享途径分析显示，糖酵解和磷酸戊糖途径显著富集（<0.001）。进一步分析糖酵解和磷酸戊糖途径抑制剂表明，这两条途径对 GCRV 复制很重要。由于肾脏是 5 种组织中受影响最大的器官，我们分析了肾脏中的丁酸盐代谢，结果表明，丁酸盐代谢中差异表达的蛋白质和代谢物大部分与 TCA 循环有关。进一步的研究表明，富马酸是 TCA 循环中的一种中间产物，可显著抑制 CIK 细胞中的 GCRV 复制（<0.001），这种抑制作用可能与其诱导干扰素系统的激活有关。在 GCRV 感染期间，向饲料中添加富马酸可使草鱼幼鱼的存活率提高 19.60%，并保护感染 GCRV 的组织，使其成为一种有潜力的抗 GCRV 饲料添加剂。本研究结果为草鱼 GCRV 发病机制和抗病毒策略提供了新的视角。
