目的 基于网络药理学和肠道菌群探究五苓胶囊抗慢性肾损伤（CKI）的作用机制。方法 利用中药系统药理学数据库分析平台（TCMSP）对五苓胶囊中泽泻、猪苓、茯苓、白术、肉桂的活性成分进行筛选，并通过UniProt和Swiss Target Prediction数据库，对筛选所得活性成分进行作用靶点预测；通过GeneCards与OMIM数据库获得慢性肾损伤的疾病靶点与肠道菌群靶点；通过STRING数据库以及Cytoscape软件构建蛋白质-蛋白质相互作用（PPI）网络及“中药-成分-靶点-疾病”网络；通过DAVID数据库对关键靶点进行基因本体（GO）富集分析、京都基因与基因组百科全书（KEGG）通路富集分析。采用5/6肾切除建立CKI大鼠模型，并分为对照组、模型组、阳性药(依那普利0.5 mg·kg^-1)组及五苓胶囊低、高剂量(124、498 mg·kg^-1)组。造模成功后，各给药组ig相应的药物，连续4周。治疗结束后，分别收集各组大鼠粪便，进行肠道菌群的特征分析。结果 通过筛选共得到53种有效成分和80个药物-疾病-肠道菌群共同靶点，GO功能富集分析共得到488个相关条目，其中生物过程（BP）321条，细胞组成（CC）48条，分子功能（MF）119条。KEGG富集共得到143条信号通路。网络药理学结果表明，五苓胶囊抗CKI的作用机制与氧化应激及炎症反应有关。肠道菌群结果表明，模型组大鼠肠道菌群发生明显改变；五苓胶囊治疗后的肠道菌群物种多样性和丰富度均有所恢复。经五苓胶囊治疗后，厚壁菌门（Firmicutes）/拟杆菌门（Bacteroidetes）(F/B)值显著下降。动物实验结果表明，五苓胶囊可改善由CKI造成的肾脏多部位的病理性损伤，降低血清中肌酐（Scr）、尿素氮（BUN）、丙二醛（MDA）水平，升高肾脏保护因子超氧化物歧化酶（SOD）的水平。结论 五苓胶囊抗CKI的作用可能与调控前列腺素内过氧化物合酶2（PTGS2）、表皮生长因子受体（EGFR）等炎症、氧化应激靶点以及重塑肠道菌群结构有关，其机制可能涉及“肠-肾轴”的多环节调控，可为五苓胶囊深入研究提供方向。

Objective To explore the mechanism of Wuling Capsules against chronic kidney injury（CKI） based on network pharmacology and gut microbiota. Methods The active components of Alisma orientale（Ze Xie）, Polyporus umbellatus（Zhu Ling）, Poria cocos（Fu Ling）, Atractylodes macrocephala（Bai Zhu）, and Cinnamomum cassia（Rou Gui） in Wuling Capsules were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform（TCMSP）. The targets of the screened active components were predicted via the UniProt and Swiss Target Prediction databases. Disease targets related to CKI and gut microbiota-associated targets were obtained from the GeneCards and OMIM databases. Protein-protein interaction（PPI） networks and “herb-component-target-disease” networks were constructed using the STRING database and Cytoscape software. Gene Ontology（GO） enrichment analysis and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis of key targets were performed using the DAVID database. A rat model of CKI was established by 5/6 nephrectomy, and the rats were divided into the model group, normal group, positive drug group, and low-dose and high-dose Wuling Capsule groups. After successful modeling, rats in the Wuling Capsule groups were administered Wuling Capsules at low dose(124 mg·kg^-1) and high dose(498 mg·kg^-1) by ig, while those in the positive drug group were given enalapril(0.5 mg·kg^-1) by ig. At the end of treatment, fecal samples were collected from rats in each group for characteristic analysis of gut microbiota. The mechanism of Wuling Capsules in treating CKI was revealed through validation by gut microbiota analysis and animal experiments. Results A total of 53 active components and 80 common targets（drug-disease-gut microbiota） were identified. GO enrichment analysis yielded 488 GO terms, including 321 biological process terms, 48 cellular component terms, and 119 molecular function terms. KEGG enrichment analysis identified 143 signaling pathways. Network pharmacology results indicated that the mechanism of Wuling Capsules against CKI was associated with oxidative stress and inflammatory response. Gut microbiota analysis showed that the gut microbiota composition of rats in the model group changed significantly, while the species diversity and richness of gut microbiota in the Wuling Capsule-treated groups were restored. After treatment with Wuling Capsules, the Firmicutes/Bacteroidetes（F/B） ratio decreased significantly. Animal experiment results demonstrated that Wuling Capsules could improve multiple pathological injuries of the kidney caused by CKI, reduce the serum levels of serum creatinine（Scr）, blood urea nitrogen（BUN）, and malondialdehyde（MDA）, and increase the level of the renal protective factor superoxide dismutase（SOD）. Conclusion The renoprotective effects of Wuling Capsule against CKI may be associated with the modulation of inflammatory and oxidative stress targets（e.g., PTGS2 and EGFR） and the remodeling of gut microbiota. These mechanisms potentially involve multi-level regulation via the “gut-kidney axis”, providing directions for further in-depth research on Wuling Capsule.

R285.5

国家中医药综合改革示范区中医药科技共建项目(GZY-KJS-SD-2023-052); 国家自然科学基金项目(82505204); 烟台市校地融合发展项目(2023XDRHXMXK08); 山东省自然科学基金项目(ZR2020MH350)