小郑汤与术后膀胱内灌注相结合已被证明能改善膀胱癌患者的预后并预防复发。然而，这种中草药配方的疗效机制仍不清楚。本研究旨在利用基于网络药理学的实验，识别小郑汤的重要成分，并探究其抗膀胱癌作用和机制。从传统中药（TCM）数据库收集了小郑汤中每种草药的化学成分。采用网络药理学预测作用的靶蛋白和途径。利用疾病数据库识别与膀胱癌相关的靶基因。构建蛋白质相互作用（PPI）网络来说明与相交靶蛋白的相互作用。使用基因本体论（GO）和京都基因和基因组百科全书（KEGG）功能富集分析确定关键靶标。在分子对接预测后建立了一个化合物-靶标-途径网络。通过超高效液相色谱-四级杆飞行时间质谱（UPLC-qTOF-MS）确定的核心化学成分进行了膀胱癌细胞系的体外实验，以验证网络药理学的结论。提取了45种活性化合物，并描述了它们与传统中药（TCMs）和蛋白靶标的关系，包括7种草药、45种活性成分和557个蛋白靶标。潜在TCM靶基因与膀胱癌相关基因的交集得到322个基因。GO和KEGG分析表明这些靶标可能参与许多与癌症相关的途径。分子对接结果显示，除mandenol外，候选化合物可以与受体形成稳定的构象。对三种膀胱癌细胞系进行的体外实验表明，槲皮素和另外两种引人注目的新化合物——低甲氧基姜黄素和葛它酮，通过B细胞淋巴瘤2/Bcl-2相关X（Bcl-2/BAX）途径显著促进癌细胞凋亡，并通过糖原合酶激酶3β（GSK3β）/β-连环蛋白途径抑制增殖和迁移。通过应用网络药理学和进行体外实验，初步阐明了小郑汤对抗膀胱癌的机制，确定了其主要活性成分和靶标，为未来研究提供了科学依据。© 2023. 国际中国医学学会和BioMed Central有限公司。

The combination of Xiaozheng decoction with postoperative intravesical instillation has been shown to improve the prognosis of bladder cancer patients and prevent recurrence. However, the mechanisms underlying the efficacy of this herbal formula remain largely unclear. This research aims to identify the important components of Xiaozheng decoction and explore their anti-bladder cancer effect and mechanism using network pharmacology-based experiments.The chemical ingredients of each herb in the Xiaozheng decoction were collected from the Traditional Chinese Medicine (TCM) database. Network pharmacology was employed to predict the target proteins and pathways of action. Disease databases were utilized to identify target genes associated with bladder cancer. A Protein-Protein Interaction (PPI) network was constructed to illustrate the interaction with intersected target proteins. Key targets were identified using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment analysis. A compound-target-pathway network was established after molecular docking predictions. In vitro experiments with bladder cancer cell lines were conducted using core chemical components confirmed by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-qTOF-MS) to verify the conclusions of network pharmacology.45 active compounds were extracted, and their relationships with Traditional Chinese Medicines (TCMs) and protein targets were presented, comprising 7 herbs, 45 active compounds, and 557 protein targets. The intersection between potential TCM target genes and bladder cancer-related genes yielded 322 genes. GO and KEGG analyses indicated that these targets may be involved in numerous cancer-related pathways. Molecular docking results showed that candidate compounds except mandenol could form stable conformations with the receptor. In vitro experiments on three bladder cancer cell lines demonstrated that quercetin and two other impressive new compounds, bisdemethoxycurcumin (BDMC) and kumatakenin, significantly promoted cancer cell apoptosis through the B-cell lymphoma 2/Bcl-2-associated X (Bcl-2/BAX) pathway and inhibited proliferation and migration through the glycogen synthase kinase 3 beta (GSK3β)/β-catenin pathway.By employing network pharmacology and conducting in vitro experiments, the mechanism of Xiaozheng decoction's effect against bladder cancer was tentatively elucidated, and its main active ingredients and targets were identified, providing a scientific basis for future research.© 2023. International Society for Chinese Medicine and BioMed Central Ltd.