Exploration of the potential mechanism of the Fuzheng Xiegan decoction for treating hepatocellular carcinoma based on experimental and network pharmacological analyses

As a traditional Chinese medicine (TCM) prescription, Fuzheng Xiegan decoction (FXD) has shown protective effects on hepatocellular carcinoma (HCC). This study aimed to investigate the specific mechanism of action of FXD in HCC through in vitro experiments. Experimental and network pharmacological analyses were used to test the mechanism of FXD in HCC. Cell cycle, apoptosis, invasion, and migration assays were conducted to verify the effect of FXD on HCC cells. Ultra-high performance liquid chromatography with quadruple time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to analyze the composition of FXD. The active compounds and potential targets of FXD, as well as HCC-related genes, were obtained from public databases. The potential targets and signaling pathways were determined by protein–protein interaction (PPI), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. The expression levels of crucial target proteins of FXD against HCC were analyzed using Real Time Quantitative Polymerase Chain Reaction (RT-qPCR) and Western blotting (WB) analysis. The results showed that FXD induced the cell cycle arrest in the G1 phase, inhibited cell proliferation, induced apoptosis, and inhibited the invasion and migration of HCC cells in a dose-dependent manner. Thirty-one major bioactive ingredients of FXD were identified. The pathway enrichment analysis identified the key target of FXD and the key PI3K/Akt and MAPK/ERK pathways. The RT-qPCR and WB analysis showed that FXD decreased the expression levels of vascular endothelial growth factor A(VEGFA), fibroblast growth factor-2 (FGF2), heat shock protein 90 alpha family class A member 1 (HSP90AA1), murine double minute (MDM2), and cyclin dependent kinase 6 (CDK6) in a dose-dependent manner. Western blot analysis showed that PI3K/Akt and MAPK/ERK pathways were significantly inhibited by FXD. This study revealed the active ingredients, potential targets, and molecular mechanisms of FXD against HCC. It also provided a promising approach to uncovering the therapeutic mechanism of action of the TCM formula against diseases.