目的 通過網(wǎng)絡(luò)藥理學、分子對接以及體外實驗探討重樓皂苷I抗宮頸癌的潛在靶點和作用機制。方法 使用Swiss Target Prediction數(shù)據(jù)庫獲得重樓皂苷I作用靶點，通過GeneCards和OMIM數(shù)據(jù)庫檢索宮頸癌的潛在作用靶點。采用Venny 2.1和Cytoscape 3.10.0構(gòu)建“重樓皂苷I-交集靶點”網(wǎng)絡(luò)。通過STRING平臺構(gòu)建蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)圖篩選出重樓皂苷I抗宮頸癌的可能的核心靶點，通過富集分析對核心靶點的信號通路及生物學功能進行分析，利用分子對接方法評估重樓皂苷I與關(guān)鍵信號通路中核心靶蛋白的結(jié)合潛力。通過CCK-8法檢測宮頸癌細胞增殖能力，劃痕實驗檢測細胞的遷移能力，Western blotting檢測靶點蛋白表達水平。結(jié)果 通過數(shù)據(jù)庫得到105個重樓皂苷I作用靶點，Venn圖得到96個交集靶點，通過蛋白質(zhì)相互作用網(wǎng)絡(luò)篩選出酪氨酸蛋白激酶（SRC）、表皮生長因子受體（EGFR）、胱天蛋白酶3（CASP3）、信號傳導轉(zhuǎn)錄激活因子3（STAT3）等35個核心靶點。GO分析結(jié)果表明重樓皂苷I顯著富集于200條生物過程，40個分子功能，49個細胞組分；KEGG富集分析獲得99條通路。分子對接顯示重樓皂苷I與c-Jun氨基末端激酶（JNK）、細胞外調(diào)節(jié)蛋白激酶（ERK）、p38蛋白（p38）有較好的結(jié)合力。體外細胞實驗表明，重樓皂苷I抑制宮頸癌SiHa細胞的增殖活力和遷移能力。Western blotting結(jié)果表明，重樓皂苷I可以上調(diào)MAPK信號通路中磷酸化蛋白的表達水平。結(jié)論 重樓皂苷I可能通過作用于MAPK信號通路發(fā)揮抗宮頸癌作用。

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

湖北省中醫(yī)藥管理局中醫(yī)藥科研項目（ZY2023F075）；湖北醫(yī)藥學院“十四五”省優(yōu)勢特色學科群（生物與醫(yī)藥）資助項目（2024BMXKQT6）；武當特色中藥研究湖北省重點實驗室（湖北醫(yī)藥學院）開放課題（WDCM2024024）；十堰市引導性科研項目（24Y139）