Jujuboside B from Zizyphi spinosi semen inhibits hepatocellular carcinoma progression by targeting the PI3K/Akt signaling pathway in vitro and in vivo

Hepatocellular carcinoma (HCC) is prevalent malignant neoplasm characterized by restricted therapeutic alternatives and an unfavorable prognosis. The objective of this study is to clarify the molecular mechanisms underlying the effects of Zizyphi spinosi semen (ZSS) and its bioactive component Jujuboside B (JUB) exert anti-HCC effects. To achieve this goal, network pharmacology, bioinformatics, molecular docking, molecular dynamics simulations, surface plasmon resonance (SPR) and in vitro and in vivo experiments was employed. First, 26 bioactive components of ZSS and their corresponding 144 potential targets were identified through the SwissTargetPrediction, BATMAN-TCM and TCMSP databases. Then 105 ZSS bioactive components targets for HCC were screened out. An analysis for GO and KEGG pathways was conducted and the PI3K/AKT signaling pathway was markedly enriched. Protein-protein interaction (PPI) network analysis identified SRC, AKT1, ESR1, TP53, and EGFR as key targets. Molecular docking, molecular dynamics simulations and SPR studies showed that JUB a bioactive component of ZSS had the strongest binding affinity with AKT1, supporting its role in regulating the PI3K/Akt pathway. In vitro experiments found that JUB can inhibit the growth of HepG2 cells and induced apoptosis by upregulating Bax and downregulating Bcl-2 expression. Further experiments showed that JUB reduced the expression of p-PI3K and p-AKT. Intervention with the PI3K/AKT pathway activator Recilisib reversed the effects of JUB on HepG2 cells, indicating that JUB exerts its function on HepG2 cells via the PI3K/AKT pathway. In vivo studies using a rat HCC model showed that intervention with JUB significantly reduced tumor formation and improved liver pathology. Immunohistochemical results revealed that intervention with JUB induced apoptosis and inhibited PI3K/Akt pathway. Overall, these results of this study indicate that JUB from ZSS exerts its anti-HCC effects by inhibiting the PI3K/AKT pathway and regulating apoptosis-related proteins, demonstrating its potential for use in HCC management. • Through integrated network pharmacology and experimental validation, the bioactive component Jujuboside B (JUB) derived from Zizyphi spinosi semen (ZSS) was shown to exhibit anti-hepatocellular carcinoma activity. • JUB can bind directly to the core target AKT1. • JUB inhibits hepatocellular carcinoma proliferation and induces apoptosis both in vitro and in vivo. • The anti-HCC mechanism of JUB is mediated through the inhibition of the PI3K/AKT signaling pathway.