Integrating network pharmacology and metabolomics to explore the mechanism of Xihuang Pill in exerting anti-tumor immune effects against pancreatic ductal adenocarcinoma.

ETHNOPHARMACOLOGICAL RELEVANCE: Xihuang Pill (XHP), a traditional Chinese medicine formula, has demonstrated anti-pancreatic ductal adenocarcinoma (PDAC) efficacy in prior studies. However, its effects on tumor-associated macrophages (TAMs) and underlying mechanisms remain unclear.

AIM OF THE STUDY: This study aimed to elucidate the mechanism of XHP in exerting anti-tumor immune effects against PDAC through network pharmacology, widely targeted metabolomics, and in vitro and in vivo experiments.

MATERIALS AND

METHODS

UPLC-QTOF-MS and network pharmacology were employed to analyze the components and potential mechanisms of XHP, and molecular docking and molecular dynamics were used to analyze target-component interactions. Survival analysis and immune infiltration assessment of target were performed through public databases. The effects of XHP were evaluated in vitro and in vivo experiments. Widely targeted metabolomics was performed to identify key metabolites, and the effect of L-kynurenine (Kyn) was verified. WB and RT-qPCR were conducted to verify the molecular mechanism.

RESULTS

A total of 238 chemical components of XHP were identified. Network pharmacology analysis revealed that XHP might regulate multi-target and influence the PI3K/AKT pathway for PDAC treatment. Molecular docking and dynamics simulations indicated stable interactions between active components and targets. The target was linked to poor prognosis in PAAD and was correlated with macrophages. In vitro and in vivo experimental results showed that XHP inhibited tumor growth, reduced the serum CA19-9 level, and suppressed the proliferation and migration of PDAC cells. In addition, XHP increased M1 phenotype and inhibited the polarization to M2 phenotype. Metabolomics analysis revealed that XHP decreased Kyn levels and confirmed that Kyn inhibited Cd86 mRNA expression and upregulated the mRNA expression of Akt1 and Mtor. Furthermore, XHP also downregulated the expression of proteins related to the AKT/mTOR pathway, and it was revealed that β-boswellic acid affected the Kyn levels and macrophage polarization.

CONCLUSION

XHP may exert an anti-PDAC effect by influencing the Kyn level to regulate the tumor immune microenvironment and downregulating the AKT/mTOR signaling pathway. Kyn is a key metabolite of XHP for the treatment of PDAC.