Network pharmacology and pharmac-ological evaluation for anti-liver cancer mechanism study of Sanguisorba officinalis

Abstract

Background and Objectives: Sanguisorba officinalis (SO) is a well-known traditional Chinese medicine (TCM), commonly applied to treat complex diseases, as anticancer, antibacterial, antiviral, anti-inflammatory, anti-oxidant, and hemostatic effects. Especially, it has been reported to exert anti-tumor effect in various human cancers. However, its effect and pharmacological mechanism on hepatocellular carcinoma (HCC) remains unclear. This study, therefore, aimed to study the anti-HCC activity of SO and its potential underlying therapeutic mechanisms. Methods: In this study, network pharmacology approach was applied to characterize the underlying mechanism of SO in HCC. Active compounds and potential targets of SO, as well as related genes of HCC were obtained from the public databases, the potential targets and signaling pathways were determined by protein-protein interaction (PPI), gene ontology (GO), and pathway enrichment analysis. Subsequently, cell counting kit 8 (CCK8) and 5-Ethynyl-2’-deoxyuridine (EdU) staining, Western blot, immunohistochemistry, flow cytometry of Annexin V/PI, wound-healing and transwell invasion assay were performed to verify the anticancer effects and mechanism of SO on HCC in vitro and in vivo. Results: In this study, in vitro, SO was found to inhibit cell proliferation, induce apoptosis and down-regulate cell migration and invasion in various HCC cells in a dose-dependent manner. By using the comprehensive network pharmacology analysis, all ingredients in SO were collected from the corresponding databases, 12 active ingredients screened according to their oral bioavailability and drug-likeness index, and 258 potential targets related to HCC were predicted. Through enrichment analysis, SO was found to show its excellent therapeutic effects on HCC mainly via the PI3K/AKT signaling pathway. The in vitro and in vivo results showed that SO treatment down-regulated the phosphorylation level of PI3K and AKT in a dose-dependent manner. Meanwhile, PI3K/AKT inhibitor LY294002 promoted SO-induced apoptosis. On the contrary, PI3K/AKT activator insulin-like growth factor-1 (IGF-1) attenuated lethality of SO in HepG2 cells. Finally, the in vivo experiments further validated that SO exhibited its prominent therapeutic effects on HCC. Conclusion: Taken together, this study revealed the anti-tumor effects of SO and its potential underlying therapeutic mechanisms on HCC. Additionally, this study suggested combination of network pharmacology prediction with experimental validation may offer a useful tool to characterize the molecular mechanism of traditional Chinese medicine (TCM) on HCC.