目的 基于网络药理学预测桂枝茯苓丸治疗子宫内膜异位症（EMS）的潜在作用靶点与信号通路，并通过分子对接和动物实验进行机制验证。方法 利用中药系统药理学数据库与分析平台（TCMSP）和SwissTargetPrediction数据库筛选桂枝茯苓丸（桂枝、茯苓、牡丹皮、桃仁、芍药）的活性成分及其靶点；通过GeneCards、OMIM、PharmGkb、TTD和DrugBank数据库获取EMS疾病靶点；取交集获得桂枝茯苓丸-EMS共同靶点（R语言可视化）。构建“活性成分-靶点”调控网络和STRING蛋白质-蛋白质相互作用（PPI）网络；运用CytoNCA进行PPI网络拓扑分析筛选核心靶点；对核心靶点进行基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集分析；对“成分-靶点”网络中的关键活性成分与核心靶点进行分子对接验证。建立大鼠自体移植EMS模型。将40只造模成功大鼠随机分为模型组、桂枝茯苓丸低、中、高剂量(0.62、1.24、2.17 g·kg^-1)组，另设10只对照组（正常大鼠）。ELISA法检测各组大鼠血清白细胞介素（IL）-6、IL-1β、肿瘤坏死因子（TNF-α）水平；苏木精-伊红（HE）染色观察大鼠子宫内膜组织病理变化；免疫组化、Western blotting检测异位内膜病灶中磷酸酯酶与张力蛋白同源物（PTEN）、磷脂酰肌醇-3-激酶（PI3K）、蛋白激酶B（Akt）蛋白表达。结果 共筛选出桂枝茯苓丸活性成分82个，预测靶点962个；EMS疾病靶点2 492个；桂枝茯苓丸与EMS共同靶点246个。核心PPI网络筛选出关键靶点164个，核心蛋白21个。GO分析显示共同靶点主要富集于化学反应、分子功能调节、信号受体结合等生物过程；KEGG富集分析提示关键通路包括TNF、PI3K/Akt、MAPK等信号通路。分子对接表明关键活性成分（55个）与核心靶点（341个取交集得164个关键靶点）具有良好结合活性。动物实验：与对照组相比，模型组大鼠血清IL-6、IL-1β、TNF-α水平显著升高（P<0.01），子宫内膜组织呈现明显炎症浸润，且PTEN、PI3K、Akt蛋白表达显著上调（P<0.01）。与模型组相比，桂枝茯苓丸各剂量组大鼠血清炎症因子水平显著降低（P<0.01），子宫内膜炎性损伤显著减轻，PTEN、PI3K、Akt蛋白表达均显著下调（P<0.01）。结论 桂枝茯苓丸可能通过调控炎症反应、细胞信号转导及血管生成等过程发挥抗EMS作用，其机制可能与抑制PTEN/PI3K/Akt信号通路的异常激活有关。

Objective To predict the potential targets and signaling pathways of Guizhi Fuling Pills in the treatment of endometriosis（EMS） based on network pharmacology, and to verify the mechanism through molecular docking and animal experiments. Methods The active components and their targets of Guizhi Fuling Pills（Cinnamomi Ramulus, Poria, Moutan Cortex, Persicae Semen, and Paeoniae Alba Radix） were screened using the TCMSP and SwissTargetPrediction databases. The disease targets of EMS were obtained from the GeneCards, OMIM, PharmGkb, TTD, and DrugBank databases. The common targets of Guizhi Fuling Pills and EMS were obtained by taking the intersection（visualized by R language）. The “active component-target” regulatory network and STRING proteinprotein interaction（PPI） network were constructed. The CytoNCA was used for PPI network topological analysis to screen the core targets. The core targets were subjected to Gene Ontology（GO） functional annotation and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis. Molecular docking was performed to verify the key active components and core targets in the “component-target” network. A rat model of autologous transplantation of EMS was established. Forty rats with successful modeling were randomly divided into the model group, low-, medium-, and high-dose Guizhi Fuling Pills treatment groups, and another 10 rats were set as the control group（normal rats）. The levels of serum IL-6, IL-1β, and TNF-α were detected by ELISA; The pathological changes of endometrial tissue were observed by HE staining; And the expressions of PTEN, PI3K, and Akt proteins in ectopic endometrial lesions were detected by immunohistochemistry and Western blotting. Results A total of 82 active components and 962 predicted targets of Guizhi Fuling Pills were screened; 2 492 disease targets of EMS were obtained; And 246 common targets of Guizhi Fuling Pills and EMS were identified. The core PPI network screened out 164 key targets and 21 core proteins. GO analysis showed that the common targets were mainly enriched in biological processes such as chemical reactions, molecular function regulation, and signal receptor binding. KEGG enrichment analysis suggested that the key pathways included TNF, PI3K/Akt, and MAPK signaling pathways. Molecular docking indicated that the key active components（55） had good binding activity with the core targets（341, with an intersection of 164 key targets）. Animal experiments: Compared with the sham operation group, the levels of serum IL-6, IL-1β, and TNF-α in the model group were significantly increased（P ＜ 0.01）, and the endometrial tissue showed obvious inflammatory infiltration, with significantly upregulated expressions of PTEN, PI3K, and Akt proteins. Compared with the model group, the levels of inflammatory factors in the serum of rats in each dose Guizhi Fuling Pills treatment group were significantly decreased（P ＜ 0.01）, the endometrial inflammatory damage was significantly alleviated, and the expressions of PTEN, PI3K, and Akt proteins were significantly downregulated（P ＜ 0.01）. Conclusion GUIZHI FULING PILLS may exert anti-EMS effects by regulating inflammatory responses, cell signal transduction, and angiogenesis, and its mechanism may be related to the inhibition of abnormal activation of the PTEN/PI3K/Akt signaling pathway.

R285.5

江西省中医药管理局科技计划项目(2024B0465)