Unraveling the Anti-Colorectal Cancer Mechanisms of Acanthopanax senticosus Polysaccharide: A Multi-omics Investigation into Gut Microbiota-Metabolism-Immunity Crosstalk

Jiaxin Jiang¹Xiwu Zhang¹Di Han¹Qichao Liang¹Le Yang²Ling Kong¹Yu Guan¹Hui Sun^1*Chang Liu¹Ye Sun²Ying Han¹Jie Zhang³Xijun Wang^1,2*

• ¹Heilongjiang University of Chinese Medicine, Harbin, China
• ²The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, China
• ³Technology Innovation Center of Wusulijiang Ciwujia, hulin, China

Background: This study aimed to systematically investigate the anti-colorectal cancer (CRC) efficacy and the underlying mechanisms of Acanthopanax senticosus polysaccharide (ASP), with a focus on its role in modulating the gut microbiota-metabolism-immune axis. Methods: A homogeneous ASP fraction was structurally characterized using HPSEC, monosaccharide composition analysis, SEM, and FT-IR. Its anti-tumor activity was evaluated in a CT-26 tumor-bearing mouse model through histopathology, tumor inhibition rate, immune organ indices, and serum cytokine (IFN-γ, TNF-α, IL-2) assays. The potential mechanisms of action were elucidated by integrating 16S rDNA sequencing of gut microbiota, determination of short-chain fatty acids (SCFAs), untargeted serum metabolomics using UPLC-Q-TOF/MS, molecular docking studies, western blot analysis of key signaling proteins, and validation through in vitro cell experiments. Results: ASP demonstrated significant dose-dependent anti-tumor activity, with the medium dose showing the highest efficacy (50.84% inhibition). It induced tumor cell apoptosis, normalized tumor-associated immune organ hypertrophy, and rebalanced pro-and anti-tumor cytokines. Metabolomics identified 12 key biomarkers, revealing that ASP primarily reversed CRC-induced disruptions in glycerophospholipid and tryptophan metabolism. Concurrently, ASP restored gut microbiota diversity, suppressed pro-inflammatory genera, and promoted beneficial bacteria and some SCFAs. Integrated correlation analysis established a robust link between microbiota remodeling and metabolic correction. Molecular docking, Western blot validation and vitro cell experiments confirmed that ASP and its regulated metabolites could inhibit the activity of PLA2/TLR4/MyD88/NF-κB signaling pathway. Conclusion: The results indicate that anti-CRC effect of ASP may be jointly regulated through multiple pathways: correcting abnormal glycerophospholipid and tryptophan metabolism in the host, restoring the homeostasis of the intestinal microbiota, increase the content of some SCFAs, and inhibiting the TLR4/NF-κB signaling pathway.