Abstract
BACKGROUND: Colorectal cancer (CRC) remains one of the most commonly diagnosed malignancies worldwide and is associated with high recurrence and poor survival outcomes, particularly in its advanced stages. Zi-Shen-Gu-Sui Decoction (ZSGSD), a traditional Chinese medicinal formula, has been clinically applied in CRC treatment and postoperative rehabilitation; however, its pharmacological mechanisms are not yet fully elucidated. This study aimed to explore the anti-CRC efficacy and molecular mechanisms of ZSGSD, with particular attention to the PI3K/AKT/mTOR signaling axis. METHODS: UPLC-Q-Orbitrap HRMS was utilized to characterize the chemical profile of ZSGSD. Network pharmacology was employed to identify key signaling pathways and bioactive compounds, and molecular docking was conducted to predict potential interactions between major constituents and PI3K/AKT/mTOR proteins. A xenograft mouse model of CRC was established to evaluate tumor suppression. Histological staining was used to assess apoptosis, and the expression of PI3K, AKT, and mTOR was quantified via RT-qPCR, Western blotting, and immunofluorescence assays. RESULTS: Sixty compounds were identified in ZSGSD, with flavonoids comprising approximately 50%. Network analysis suggested potential involvement of the PI3K/AKT/mTOR pathway, and docking results indicated favorable predicted binding between key flavonoids (including Genistein, Apigenin, and Naringenin) and proteins within this pathway. In vivo, ZSGSD treatment suppressed tumor growth in a dose-responsive fashion, with tumor inhibition rates ranging from 30.83% to 75.23% (p < 0.05). Histological evaluations revealed reduced nuclear-to-cytoplasmic ratios and enhanced apoptotic activity, as confirmed by TUNEL staining (p < 0.05). Molecular analyses showed a dose-related reduction in PI3K, AKT, and mTOR expression at the transcript and protein levels, indicating that this pathway may be modulated during ZSGSD treatment (p < 0.05). CONCLUSION: ZSGSD demonstrates notable tumor-suppressive and pro-apoptotic effects in a CRC xenograft model. These outcomes appear to be associated with modulation of the PI3K/AKT/mTOR signaling pathway, although further functional studies are required to confirm pathway specificity and clarify causal mechanisms. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s41065-026-00636-3.