目的 通过网络药理学、分子对接以及体外实验探讨重楼皂苷I抗宫颈癌的潜在靶点和作用机制。方法 使用Swiss Target Prediction数据库获得重楼皂苷I作用靶点，通过GeneCards和OMIM数据库检索宫颈癌的潜在作用靶点。采用Venny 2.1和Cytoscape 3.10.0构建“重楼皂苷I-交集靶点”网络。通过STRING平台构建蛋白质相互作用（PPI）网络图筛选出重楼皂苷I抗宫颈癌的可能的核心靶点，通过富集分析对核心靶点的信号通路及生物学功能进行分析，利用分子对接方法评估重楼皂苷I与关键信号通路中核心靶蛋白的结合潜力。通过CCK-8法检测宫颈癌细胞增殖能力，划痕实验检测细胞的迁移能力，Western blotting检测靶点蛋白表达水平。结果 通过数据库得到105个重楼皂苷I作用靶点，Venn图得到96个交集靶点，通过蛋白质相互作用网络筛选出酪氨酸蛋白激酶（SRC）、表皮生长因子受体（EGFR）、胱天蛋白酶3（CASP3）、信号传导转录激活因子3（STAT3）等35个核心靶点。GO分析结果表明重楼皂苷I显著富集于200条生物过程，40个分子功能，49个细胞组分；KEGG富集分析获得99条通路。分子对接显示重楼皂苷I与c-Jun氨基末端激酶（JNK）、细胞外调节蛋白激酶（ERK）、p38蛋白（p38）有较好的结合力。体外细胞实验表明，重楼皂苷I抑制宫颈癌SiHa细胞的增殖活力和迁移能力。Western blotting结果表明，重楼皂苷I可以上调MAPK信号通路中磷酸化蛋白的表达水平。结论 重楼皂苷I可能通过作用于MAPK信号通路发挥抗宫颈癌作用。

Objective To explore the potential targets and action mechanisms of polyphyllin I in treating cervical cancer through network pharmacology, molecular docking, and in vitro experiments. Methods The potential targets for polyphyllin I in treating diseases were obtained from the Swiss Target Prediction database. Potential targets for cervical cancer were searched in the GeneCards and OMIM databases. The “polyphyllin I – intersection target” network was constructed using Venny 2.1 and Cytoscape 3.10.0. The potential core targets of polyphyllin I against cervical cancer were screened out by constructing a PPI network map through the STRING platform. The signaling pathways and biological functions of the core targets were analyzed by enrichment analysis. The binding potential of polyphyllin I to the core target proteins in the key signaling pathways was evaluated by molecular docking. The proliferation ability of cervical cancer cells was detected by the CCK-8 method, the migration ability of cells was detected by the scratch test, and the expression levels of target proteins were detected by Western blotting. Results A total of 105 potential targets for polyphyllin I in treating cervical cancer were identified from the databases, 96 intersection targets were obtained by Venn diagram, and 35 core targets, such as SRC, EGFR, CASP3, STAT3 were screened through the protein interaction network. GO analysis showed that polyphyllin I was significantly enriched in 200 biological processes, 40 molecular functions and 49 cell components. KEGG enrichment analysis obtained 99 pathways. Molecular docking showed that polyphyllin I had good binding force with the JNK, ERK, and p38. In vitro cell experiments showed that polyphyllin I inhibited the proliferation and migration of SiHa cervical cancer cells. Western blotting analysis showed that polyphyllin I can up-regulate the expression of phosphorylated protein in MAPK signaling pathway. Conclusion Polyphyllin I may play an anti-cervical cancer role by acting on MAPK signaling pathway.

R285.5

湖北省中医药管理局中医药科研项目（ZY2023F075）；湖北医药学院“十四五”省优势特色学科群（生物与医药）资助项目（2024BMXKQT6）；武当特色中药研究湖北省重点实验室（湖北医药学院）开放课题（WDCM2024024）；十堰市引导性科研项目（24Y139）