Comprehensive characterisation of bioactive compounds in Boletus edulis as functional foods to improve muscle atrophy; through whole plant targeted metabolomics, network pharmacology, in vivo and in vitro experiments, molecular docking and molecular dynamics analysis

Ethnopharmacological significance: Boletus edulis (BE) is a naturally occurring fungus that has been traditionally used in ancient Chinese herbal medicine. It is a key component of the formula 'Shujin Pill', commonly prescribed for the treatment or relief of muscular dystrophy. However, the specific efficacy of BE within Shujin Pill or its primary active components remains unclear. Aims of the study: This study aims to elucidate the biological function and molecular mechanisms of BE in alleviating muscular atrophy in mice. We employed a comprehensive approach, integrating metabolomics, network pharmacological analysis, molecular docking, molecular dynamics simulation, and in vivo and in vitro experimental validation, to verify these effects. Materials and methods: The bioactive components in BE were quantified by UPLC-QTOF-MS/MS. To evaluate the muscle function indexes after 14 days of action of different doses of BE and to analyze the pathological changes in muscle tissue. Enabling network pharmacology to analyze the potential active components in BE for the alleviation of muscle atrophy, using computer molecular simulation for docking scores, molecular dynamics simulation to assist in the validation of the active components in BE, and in vitro experiments for the validation of the active components. Result: BE administered alone was able to slow down Lipopolysaccharide (LPS)-induced muscle atrophy. 996 non-volatile components were detected in BE by metabolomics, and GAPDH, TP53, AKT1, TNF-alpha and IL-6 were more strongly associated with muscle atrophy by using web-based pharmacological analyses. Folic acid, Cycloartenol and Sesamin active ingredients have greater potential to treat or alleviate muscle atrophy, molecular docking, molecular dynamics detected that Sesamin and AKT both have high binding energy, in vitro using C2C12 skeletal muscle cells to verify the efficacy of Sesamin and BE, found that in the presence of the LY294002 (PI3K inhibitor) and GSK21417 (AKT inhibitor) treatment conditions, the elimination of the up- regulation of the PI3K/AKT signaling pathway by Sesamin and BE and loss of biological efficacy. It suggests that BE may slow down or treat muscle atrophy through the PI3K/AKT signaling pathway, in which Sesamin plays a major role. Meanwhile BE and Sesamin were able to enhance the antioxidant level of C2C12 skeletal muscle cells.