Abstract
To explore the specific effects and potential mechanisms of Aitongxiao Formula (ATXF) in treating HCC. Network pharmacology was first used to predict the potential targets and pathways of ATXF against liver cancer. In vitro and in vivo experiments were conducted to evaluate the effects of ATXF. The CCK8 assay and other experiments were used to assess the inhibitory effect of ATXF on cells, and the Transwell assay was used to verify the impact on cell migration and invasion. Xenograft models of liver cancer were successfully established by subcutaneously inoculating liver cancer cells, and the actual effects of different concentrations of ATXF in vivo were evaluated. Advanced technologies such as PCR and IHC were used to detect changes in core targets and other key targets in the pathways. ATXF inhibited the proliferation and metastasis behavior of liver cancer cells. It also suppressed the growth of liver cancer tumors in mice. ATXF regulated pathological changes in tumor tissues, including proliferation marker Ki67 and apoptosis marker caspase3. Network pharmacology techniques was predicted 13 effective components and 119 potential targets of ATXF. The main active components were quercetin, sitosterol, luteolin, and eugenol. These components may precisely targeted key molecules such as IL6, TP53, AKT1, TNF, IL1B, and EGFR. By examining the tumor tissues in mice, it was found that ATXF can regulate the changes in these six indicators and modulate the main proteins of the MAPK/ERK signaling pathway, thus exerting its therapeutic effect. ATXF effectively inhibits the development of hepatocellular carcinoma both in vitro and in vivo. The unique complex network of multi-component, multi-target, and multi-pathway interactions provides valuable clues and new insights for further revealing the mechanisms of action of this formula.