青鳍络纹鲮（Hexagrammos otakii）是一种海洋鱼类，石蒜素（ART）由于其免疫调节活性，被广泛用作饲料添加剂。然而，ART在海洋鱼类中的免疫调节机制尚不明确。本研究全面探究了ART改善青鳍络纹鲮肠道免疫疾病（IID）的效果，并探讨了其潜在机制。通过传统中药系统药理学（TCMSP）数据库筛选了ART的靶点。在Uniprot数据库中鉴定了八个ART的靶点，并通过Drugbank、Genecards、OMIM和PHARMGKB数据库筛选出与IID相关的1419个靶蛋白。基因本体论（GO）和京都基因与基因组百科全书（KEGG）通路分析结果表明，通过缺氧诱导因子-1（HIF-1）信号通路，ART可能通过调节细胞应对应激、缺氧、炎症和血管内皮生长因子刺激来产生免疫保护作用。分子对接结果表明，ART含有一种活性成分和三种交叉靶点，分别与缺氧诱导因子1α（HIF1-α）、转录因子p65（RELA）和血管内皮生长因子A（VEGF-A）结合。此外，建立了ART饲料模型，评估ART对青鳍络纹鲮肠道免疫保护作用。D48组在被温伤弧菌感染后显示小的肠道结构变化。ART补充饲料改善了青鳍络纹鲮肠道中的总超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和谷胱甘肽过氧化酶（GSH-Px）的水平，并降低了肠道中的丙二醛（MDA）含量。青鳍络纹鲮肠道中转录因子p65、HIF1-α、VEGF-A、Cyclin D1、基质金属蛋白酶9（MMP9）、单核细胞趋化蛋白-1（MCP-1）、肿瘤坏死因子α（TNF-α）和白细胞介素6（IL-6）的表达在饲料中加入ART后下调。本研究结果表明，青鳍络纹鲮饲料中的ART通过SOD2/核-因子-kB（NFkB）/HIF1-α/VEGF-A通路改善了抗氧化剂和免疫功能，优化了肠道结构，并增强了对温伤弧菌的抵抗力。本研究集成了药理学分析和实验验证，揭示了ART对IID的作用机制，为改善青鳍络纹鲮的IID提供了线索。(原文版权所有 2023 Gu, Wang, Zhan, Wei, Shi, Cui, Peng, Han, Li, Chen, Xue and Wang)

Artemisinin (ART) is very common as a diet additive due to its immunoregulatory activities. Nonetheless, the immunoregulatory mechanism of ART in marine fish remains unknown. This study comprehensively examined the effects and explored the potential mechanism of ART ameliorating intestinal immune disease (IID) in fat greenlings (Hexagrammos otakii).The targets of ART were screened using the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Here, eight putative targets of ART were collected and identified with the Uniprot database, and 1419 IID-associated target proteins were filtered through the Drugbank, Genecards, OMIM, and PHARMGKB Databases. The results of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways point out that ART may have immunoprotective effects by regulating cellular responses to stress, hypoxia, inflammation, and vascular endothelial growth factor stimulus through the hypoxia-inducible factor 1 (HIF-1) signaling pathway. The findings of molecular docking indicated that ART contains one active ingredient and three cross-targets, which showed a kind combination with hypoxia-inducible factor 1-alpha (HIF1-a), transcription factor p65 (RELA), and vascular endothelial growth factor A (VEGF-A), respectively. Furthermore, an ART feeding model was established to assess the ART's immunoprotect effect on the intestine of H.otakii in vivo. The D48 group showed smaller intestinal structural changes after being challenged by Edwardsiella tarda. The supplementation of ART to the diet improved total superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) and reduced the malondialdehyde (MDA) in intestine of H. otakii. The expression of transcription factor p65, HIF1-α, VEGF-A, cyclin D1, matrix metalloprotease 9 (MMP9), monocyte chemoattractant protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) was decreased after dietary ART in the intestinal of H. otakii.The present results demonstrated that dietary ART improved antioxidants and immunity, optimized the intestinal structure, and increased resistance to E. tarda through the SOD2/nuclear-factor-kappa- B (NFkB)/HIF1-a/VEGF-A pathway in the intestinal tract of H.otakii. This study integrated pharmacological analysis and experimental validation and revealed the mechanism of ART on IID, which provides insight into the improvement of IID in H. otakii.Copyright © 2023 Gu, Wang, Zhan, Wei, Shi, Cui, Peng, Han, Li, Chen, Xue and Wang.