基于网络药理学和实验验证探讨桦褐孔菌三萜抗宫颈癌HeLa细胞的作用机制

Study on the Mechanism of Triterpenoids from Inonotus Obliquus Against Gastric Cancer HeLa Cell Based on Network Pharmacology and Experimental Validation

关键词:  桦褐孔菌三萜  HeLa细胞  网络药理学  分子对接  PI3K/Akt通路  HeLa裸鼠模型

Key words:  triterpenoids from Inonotus obliquus  HeLa cell  network pharmacology  molecular docking  PI3K/Akt pathway  HeLa nude mouse model

摘要:

目的 利用网络药理学和分子对接及体内验证实验探讨桦褐孔菌三萜成分抗宫颈癌HeLa细胞的潜在作用机制。方法 基于文献研究结合数据库筛选桦褐孔菌三萜的主要活性成分，寻找活性成分以及HeLa细胞交集靶点，进行PPI网络构建，GO和KEGG富集分析，分子对接进行验证。建立HeLa裸鼠模型，桦褐孔菌三萜灌胃给药，裸鼠处死后采用Western blotting检测指标。结果 经过筛选得到15个符合条件的活性成分的274个相关联靶点。与2 046个HeLa细胞相关靶点取交集，获得104个桦褐孔菌三萜抗HeLa细胞的潜在靶点。通过“药物-成分-靶点-疾病”网络分析，PIK3CA、MAPK3、MDM2、AR、CCND1可能是桦褐孔菌三萜纯化物抗HeLa细胞的潜在靶点。KEGG富集分析结果显示，PI3K/Akt信号通路、Ras信号通路、MAPK信号通路等多个信号通路可能是桦褐孔菌三萜抗HeLa细胞的潜在作用途径。分子对接结果显示，核心成分与关键靶点均有较强的潜在结合能力。体内实验结果表明桦褐孔菌三萜可抑制模型小鼠肿瘤生长，调节模型小鼠的免疫功能，促进模型小鼠肿瘤凋亡，抑制PI3K和AKT的磷酸化。结论 基于网络药理学、分子对接和体内验证方法，证明了桦褐孔菌三萜通过多组分、多靶点发挥抗HeLa细胞的作用，为桦褐孔菌的进一步研发及其抗肿瘤作用机制的探讨提供参考。

Abstract:

OBJECTIVE To explore the potential mechanism of anti-HeLa cell of triterpenoids from Inonotus obliquus by network pharmacology, molecular docking and in vivo verification experiments. METHODS Based on literature research and database, the main active components of triterpenoids from Inonotus obliquus were screened, and the active components and HeLa cell intersection targets were found. PPI network was constructed, GO and KEGG enrichment analysis were carried out, and molecular docking was verified. HeLa cell nude mice model was established, and Inonotus obliquus triterpenoids were administered by gavage. The indexes were detected and detected by Western blotting after euthanizing nude mice. RESULTS After screening, 274 associated targets of 15 qualified active components were obtained. Intersecting with 2 046 HeLa cell-related targets, 104 potential anti-HeLa cell targets of Inonotus obliquus triterpenoids were obtained. Through the "drug-component-target-disease" network analysis, PIK3CA, MAPK3, MDM2, AR and CCND1 might be potential targets for anti-HeLa cell of triterpenoids purified from Inonotus obliquus. The results of KEGG enrichment analysis showed that multiple signal pathways such as PI3K/Akt signal pathway, Ras signal pathway and MAPK signal pathway might be the potential anti-HeLa cell pathway of triterpenoids in Inonotus obliquus. The results of molecular docking showed that the core components and key targets had strong potential binding ability. The in vivo test results showed that Inonotus obliquus triterpenoids could inhibit tumor growth, regulate immune function, promote tumor apoptosis and inhibit PI3K and AKT phosphorylation in model mice. CONCLUSION Based on the methods of network pharmacology, molecular docking and in vivo verification, it is proved that the triterpenoids of Inonotus obliquus play the role of anti-HeLa cell through multi-components and multi-targets, which lays a foundation for further research and development of Inonotus obliquus and the discussion of its anti-tumor mechanism.