Exploring the Potential Mechanism of Anti-aging Effects of Spray-dried Mulberry Leaf Phenolics Based on UPLC-Q-Exactive-MS/MS and Network Pharmacology

Objective: Ultra-high performance liquid chromatography-quadrupole-electrostatic orbitrap mass spectrometry (UPLC-Q-Exactive-MS/MS) and network pharmacology methods were used to study the metabolic absorption and potential biological activity of spray-dried mulberry leaves. Methods: UPLC-Q-Exactive-MS/MS was used to detect the phenolic components of spray-dried mulberry leaves in simulated in vitro digestion, intestinal microbial fermentation and in vivo metabolic processes. Based on the detected phenolic components, the corresponding material targets were found through the database analysis platform Swiss Target and Pharmmapper, and the aging targets were obtained by searching the HAGR and Genecards databases. The PPI (Protein-Protein Interaction) network was constructed using String11.5 database and Cytoscape 3.9.1 software. DAVID 6.8 was used for gene ontology (GO) annotation and KEGG (Kyoto Encyclopedia of Genes and Genomes Analysis) pathway enrichment analysis to analyze core targets and action pathways, and then the "components-targets-pathway" network was constructed. Results: The phenolic compounds of spray-dried mulberry leaves mainly underwent hydrolysis or deglycosidation during the gastrointestinal digestion stage. After being absorbed, these substances mainly underwent phase Ⅰ and Ⅱ reactions such as reduction, hydroxylation, glucuronidation and sulfation in the body. The prototype compounds and metabolites of phenolic compounds in spray-dried mulberry leaves co-existed in the blood circulation. Through the network pharmacology method, 476 intersection targets of phenolic substances and aging were screened out. After constructing PPI, 47 core targets were screened out. The top ten targets were GAPDH (glyceraldehyde-3-phosphate dehydrogenase ), ALB (albumin), AKT1 (protein kinase B1), INS (insulin), ACTβ (β-actin), TP53 (tumor protein p53), TNF (tumor necrosis factor), IL6 (interleukin 6), EGFR (epidermal growth factor receptor), IL1β (interleukin 1β). KEGG results showed that phenolic compounds of spray-dried mulberry leaves might play an anti-aging role by regulating Wnt and TGF-β signaling pathways. Conclusion: The phenolic compounds of spray-dried mulberry leaves may play an anti-aging role through multi-component, multi-target and multi-pathway.