目的 基于模式生物斑马鱼结合网络药理学，研究枳壳的活性成分水合橙皮内酯增强免疫作用及作用机制。方法 建立2种斑马鱼免疫低下模型：以雷帕霉素建立尾部中性粒细胞数目减少模型，以酒石酸长春瑞滨建立静脉血管中巨噬细胞荧光强度减弱模型，检测水合橙皮内酯（10、20、50、100、200 μg·mL^-1）各给药组斑马鱼尾部中性粒细胞数目、斑马鱼尾部静脉血管中巨噬细胞荧光强度，计算中性粒细胞增长率和巨噬细胞改善率。基于网络药理学预测水合橙皮内酯增强免疫作用的机制，使用 Pharm Mapper、中药系统药理学数据库与分析平台（TCMSP）进行水合橙皮内酯靶点筛选，运用Gene Cards数据库和 OMIM数据库检索增强免疫相关靶点，运用 Venny2.1将上述两组靶点取交集后导入 String数据库进行蛋白质-蛋白质相互作用（PPI）分析，进行基因本体（GO）功能富集分析和京都基因与基因组百科全书（KEGG）通路富集分析，在 Cytoscape3.9.1软件中构建水合橙皮内酯的增强靶点 PPI网络，并以节点度值的中位数筛选关键靶点。结果 与模型组相比，水合橙皮内酯可显著增加斑马鱼尾部中性粒细胞数目（P＜0.01、0.001）、静脉血管中巨噬细胞荧光强度（P＜0.05），证明水合橙皮内酯具有抗雷帕霉素、酒石酸长春瑞滨引起的免疫力低下作用，呈剂量相关性。水合橙皮内酯增强免疫的作用机制可能与 AKT1、EGFR、SRC、MMP9、ESR1等关键靶点及 PI3K-Akt、Rap1、AGE-RAGE等信号通路有关。结论 水合橙皮内酯具有增强免疫作用，可能通过调控PI3K-Akt、Rap1、AGE-RAGE等信号通路发挥作用。

Objective To investigate the immunoenhancing effects and mechanisms of action of meranzin hydrate (MH), an active component of Aurantii Fructus, based on the model organism zebrafish in combination with network pharmacology.Methods Two types of immunocompromised zebrafish models were established: One using rapamycin to reduce the number of neutrophils in the tail, and another using vinorelbine to diminish the fluorescence intensity of macrophages in tail vein vessels. The neutrophil count in the tail and the fluorescence intensity of macrophages in tail vein vessels were measured in each treatment group of MH (10, 20, 50、 100, and 200 μg·mL^-1), and the growth rate of neutrophils (%) and the improvement rate of macrophages (%) were calculated. The mechanism by which MH enhances immune function was predicted using network pharmacology, with target screening for MH conducted using PharmMapper and the TCMSP database. Immune-related targets were retrieved from the Gene Cards and OMIM databases. Intersection of these target sets was analyzed for protein interactions in the String Database, followed by GO and KEGG analyses. A PPI network of MH's immunomodulatory action was constructed in Cytoscape 3.9.1, with key targets identified based on the median degree value of nodes.Results Compared to the model group, MH significantly increased both the neutrophil count (P < 0.01, 0.001) in the tail and the fluorescence intensity(P < 0.05) of macrophages in tail vein vessels, demonstrating its dose-dependent protective effects against immune suppression induced by rapamycin and camptothecin. The immunoenhancing mechanism of MH may be associated with key targets such as AKT1, EGFR, SRC, MMP9, ESR1, and signaling pathways including PI3K-Akt, Rap1, and AGE-RAGE.Conclusion MH enhances immune function, potentially through the regulation of PI3K-Akt, Rap1, and AGERAGE signaling pathways.

R285.5

山东省科技型中小企业创新能力提升工程项目（2023TSGC0271）；山东省自然科学基金联合基金项目（ZR2021LZY021）；山东省重点研发计划项目（2021CXGC010511-01-005）