金丝桃苷抗肝癌的作用机制探讨

Exploration of the Mechanism of Action of Hyperoside in Treating Liver Cancer

摘要: 目的：本研究采用网络药理学和细胞实验探讨金丝桃苷抗肝癌的分子机制。方法：通过网络药理学数据库预测金丝桃苷的作用靶点，使用疾病靶点数据库获取肝癌相关靶点，取交集后得到金丝桃苷作用于肝癌的靶点。使用Cytoscape软件构建PPI网络，筛选出核心作用靶点，采用在线注释及可视化整合分析工具(DAVID)进行GO和KEGG通路富集分析。通过CCK-8实验检测金丝桃苷对Huh7细胞活力的影响；平板克隆实验检测金丝桃苷对Huh7细胞长期增殖的影响；Calcein AM/PI实验检测金丝桃苷对Huh7细胞活死的影响；流式细胞仪检测金丝桃苷对Huh7细胞凋亡和周期的影响。结果：通过网络药理学预测到金丝桃苷作用于肝癌的171个相关靶点，并筛选出核心靶基因包括AKT1、TNF、Caspase3、INS及ALB等。KEGG通路富集分析得到主要信号通路包括癌症信号通路、AGE-RAGE信号通路、ErbB信号通路、VEGF信号通路、PI3K-AKT信号通路和HIF-1信号通路等。细胞实验显示，与空白组比较，金丝桃苷200、400 μmol·L⁻¹能降低肝癌Huh7细胞的活力(P < 0.05)，200、400 μmol·L⁻¹高浓度的金丝桃苷可抑制肝癌细胞的克隆形成；但金丝桃苷并不诱导肝癌Huh7细胞凋亡和周期阻滞。结论：金丝桃苷具有一定的抑制肝癌Huh7细胞增殖的活性，网络药理学结果表明其可能通过AKT1、TNF、Caspase3和EGFR等关键靶点和AGE-RAGE、ErbB、VEGF、PI3K-AKT和HIF-1等信号通路参与调控肝癌的发展进程。

Abstract: Objective: This study aimed to investigate the effect and molecular mechanism of hyperoside against liver cancer using network pharmacology and in vitro cellular experiments. Methods: The action targets of hyperoside were predicted using network pharmacology databases, while liver cancer-related targets were obtained from disease target databases. The overlapping targets between hyperoside and liver cancer were identified. A protein-protein interaction (PPI) network was constructed using Cytoscape software, and core targets were screened. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed using the Database for Annotation, Visualization and Integrated Discovery (DAVID). The effect of hyperoside on Huh7 cell viability was assessed by CCK-8 assay; long-term proliferation was evaluated by colony formation assay; cell viability and death were detected by Calcein AM/PI staining; and apoptosis and cell cycle distribution were analyzed by flow cytometry. Results: A total of 171 potential targets of hyperoside against liver cancer were predicted through network pharmacology, and the core target genes included AKT1, TNF, Caspase3, INS, and ALB. KEGG pathway enrichment analysis revealed that the main signaling pathways involved included pathways in cancer, the AGE-RAGE signaling pathway, the ErbB signaling pathway, the VEGF signaling pathway, the PI3K-AKT signaling pathway, and the HIF-1 signaling pathway. In vitro cellular experiments showed that compared with the control group, hyperoside at concentrations of 200 and 400 μmol·L⁻¹ significantly reduced the viability of Huh7 cells (P < 0.05). Higher concentrations of hyperoside (200 and 400 μmol·L⁻¹) inhibited colony formation of liver cancer cells. However, hyperoside did not induce apoptosis or cell cycle arrest in Huh7 cells. Conclusion: Hyperoside exhibits certain inhibitory activity against the proliferation of Huh7 liver cancer cells. Network pharmacology results suggest that it may participate in regulating the progression of liver cancer through key targets such as AKT1, TNF, Caspase3, and EGFR, as well as signaling pathways including AGE-RAGE, ErbB, VEGF, PI3K-AKT, and HIF-1.