目的 基于网络药理学及分子对接技术探究消痔丸治疗痔病的作用机制，并结合体外实验初步明确其药效物质基础。方法 选取消痔丸中主要活性成分，利用Swiss Target prediction、TCMSP预测成分靶点；通过GeneCards等数据库获取痔病相关靶点；借助STRING 12.0构建交集靶点的蛋白质相互作用（PPI）网络并筛选核心靶点；利用Metascape进行交集靶点的生物信息学分析，通过Cytoscape 3.9.1软件构建网络图；采用Schrödinger2020 Maestro12.4进行分子对接；MTS法检测细胞存活率；Griess法检测一氧化氮（NO）含量；Transwell小室法检测细胞血管通透性；细胞划痕实验检测细胞迁移率；ELISA法检测因子和蛋白含量；采用钙离子荧光法测定对五羟色胺受体2A（HTR2A）、瞬时受体电位A1（TRPA1）、瞬时受体电位香草醛亚家族1（TRPV1）的拮抗作用；采用凝血因子分析仪测定凝血四项，比浊法测定血小板聚集率。结果 筛选出毛蕊异黄酮葡萄糖苷、山柰酚、丹皮酚、白术内酯II等34个主要活性成分，获得81个成分-疾病交集靶点，确定15个核心靶点，涉及158条信号通路。分子对接显示核心成分与靶点有较好的结合作用。体外实验表明，24个化合物显著抑制RAW264.7释放NO、肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）；8个化合物降低HUVEC细胞通透性，上调血管内皮钙黏蛋白（VE-Cadherin）、闭合蛋白-5（Claudin5）、紧密连接蛋白1（ZO-1）表达；15个化合物促进人真皮成纤维细胞（HDF）的增殖与迁移，提高表皮细胞生长因子（EGF）、转化生长因子β1（TGF-β1）、血管内皮生长因子（VEGF）水平；8、11、6个化合物分别拮抗HTR2A、TRPA1及TRPV1受体；5个化合物显著延长凝血酶时间（TT）、活化部分凝血酶原时间（APTT）、凝血酶原时间（PT），6个化合物增加纤维蛋白原（FIB）含量，7个化合物可促进血小板聚集（P＜0.05、0.01、0.001）。结论 消痔丸通过抗炎消肿、促进创面愈合、止痛、止血等多重途径发挥治疗痔病的作用。

Objective To investigate the mechanism of Xiaozhi Pills in treatment of hemorrhoids based on network pharmacology and molecular docking, and to preliminarily identify its material basis through in vitro experiments. Methods The main active components of Xiaozhi Pills were selected, and the targets were predicted using Swiss Target Prediction and TCMSP. Hemorrhoid-related targets were obtained from databases such as GeneCards. PPI network of the intersecting targets was constructed using STRING 12.0, and core targets were screened. Bioinformatics analysis of the intersecting targets was performed using Metascape, and the network diagram was constructed using Cytoscape 3.9.1. Molecular docking was carried out using Schrödinger 2020 Maestro 12.4. Cell viability was assessed using the MTS assay, NO content was measured by Griess assay, vascular permeability was assessed using the Transwell assay, and cell migration rate was evaluated using the scratch wound healing assay. The levels of cytokines and proteins were measured by ELISA. Antagonistic effects on HTR2A, TRPA1, and TRPV1 were determined by calcium ion fluorescence assay. Four coagulation parameters were measured using a coagulation analyzer, and platelet aggregation rate was determined by turbidimetric method. Results 34 main active components such as genistein glucoside, kaempferol, paeonol, atractylenolide II were screened, yielding 81 component-disease intersecting targets, and 15 core targets were identified, involving 158 signaling pathways. Molecular docking showed good binding affinity between core components and targets. In vitro experiments showed that 24 compounds significantly inhibited the release of NO, TNF-α, and IL-1β from RAW264.7 cells. 8 Compounds reduced HUVEC cell permeability and upregulated the expression of VE-cadherin, Claudin-5, and ZO-1. 15 Compounds promoted the proliferation and migration of HDF and increased the levels of EGF, TGF-β1, and VEGF. 8, 11, and 6 compounds antagonized HTR2A, TRPA1, and TRPV1 receptors, respectively. 5 Compounds significantly prolonged TT, APTT, and PT. 6 Compounds increased FIB content, and 7 compounds promoted platelet aggregation (P < 0.05, 0.01, 0.001). Conclusion Xiaozhi Pills exert therapeutic effects on hemorrhoids through multiple pathways including anti-inflammation, swelling reduction, wound healing promotion, pain relief, and hemostasis.

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