目的 通過(guò)網(wǎng)絡(luò)藥理學(xué)、分子對(duì)接以及體外實(shí)驗(yàn)探討重樓皂苷I抗宮頸癌的潛在靶點(diǎn)和作用機(jī)制。方法 使用Swiss Target Prediction數(shù)據(jù)庫(kù)獲得重樓皂苷I作用靶點(diǎn)，通過(guò)GeneCards和OMIM數(shù)據(jù)庫(kù)檢索宮頸癌的潛在作用靶點(diǎn)。采用Venny 2.1和Cytoscape 3.10.0構(gòu)建“重樓皂苷I-交集靶點(diǎn)”網(wǎng)絡(luò)。通過(guò)STRING平臺(tái)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(diǎn)，通過(guò)富集分析對(duì)核心靶點(diǎn)的信號(hào)通路及生物學(xué)功能進(jìn)行分析，利用分子對(duì)接方法評(píng)估重樓皂苷I與關(guān)鍵信號(hào)通路中核心靶蛋白的結(jié)合潛力。通過(guò)CCK-8法檢測(cè)宮頸癌細(xì)胞增殖能力，劃痕實(shí)驗(yàn)檢測(cè)細(xì)胞的遷移能力，Western blotting檢測(cè)靶點(diǎn)蛋白表達(dá)水平。結(jié)果 通過(guò)數(shù)據(jù)庫(kù)得到105個(gè)重樓皂苷I作用靶點(diǎn)，Venn圖得到96個(gè)交集靶點(diǎn)，通過(guò)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(zhǎng)因子受體（EGFR）、胱天蛋白酶3（CASP3）、信號(hào)傳導(dǎo)轉(zhuǎn)錄激活因子3（STAT3）等35個(gè)核心靶點(diǎn)。GO分析結(jié)果表明重樓皂苷I顯著富集于200條生物過(guò)程，40個(gè)分子功能，49個(gè)細(xì)胞組分；KEGG富集分析獲得99條通路。分子對(duì)接顯示重樓皂苷I與c-Jun氨基末端激酶（JNK）、細(xì)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(xì)胞實(shí)驗(yàn)表明，重樓皂苷I抑制宮頸癌SiHa細(xì)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(hào)通路中磷酸化蛋白的表達(dá)水平。結(jié)論 重樓皂苷I可能通過(guò)作用于MAPK信號(hào)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(xiàng)目（ZY2023F075）；湖北醫(yī)藥學(xué)院“十四五”省優(yōu)勢(shì)特色學(xué)科群（生物與醫(yī)藥）資助項(xiàng)目（2024BMXKQT6）；武當(dāng)特色中藥研究湖北省重點(diǎn)實(shí)驗(yàn)室（湖北醫(yī)藥學(xué)院）開(kāi)放課題（WDCM2024024）；十堰市引導(dǎo)性科研項(xiàng)目（24Y139）