Integrated strategy of serum metabolomics, network pharmacology and experimental validation on revealing the bioactive metabolites and mechanism of Lycium ruthenicum Murr. against colorectal carcinoma

Lycium ruthenicum Murr. (LR) has attracted significant attention for its potential in combating colorectal carcinoma (CRC). However, the identification of key bioactive metabolites and the elucidation of their molecular mechanisms remain elusive. The study aimed to identify the bioactive metabolites profile in vivo and elucidate the mechanisms underlying their anti-CRC effects. Initially, a UHPLC-Q-Exactive-MS technique was integrated with a feature-based molecular networking approach to annotate 26 prototype metabolites in rat serum. Subsequently, a preliminary evaluation of the anti-CRC effect of the serum was conducted. Thereafter, network pharmacology, molecular docking, and molecular dynamics were employed to investigate the potential mechanisms and to screen for bioactive metabolites. Following this, the signal pathways of the primary bioactive metabolites were validated through cell experiments. Our findings suggested that the candidate targets of those prototypes' metabolites were implicated in the PI3K/Akt pathways. Furthermore, cell experiments demonstrated that LR-containing serum could inhibit the proliferation, Apoptosis, and migration of HCT-116 cells. Molecular docking and dynamics analyses indicated significant binding affinity and stability between the core targets and bioactive metabolites, such as luteolin and esculin. The bioactive metabolites of LR are potential material Bases for anti-CRC via the PI3K/Akt signalling pathway, offering new insights for the functional development and utilisation of LR.

Colorectal carcinoma; Feature-based molecular network; Lycium ruthenicum Murr.; Metabolomics; Network pharmacology.