Mining and validation of anti-inflammatory peptides from white tea based on network pharmacology and molecular docking

【Objective】 This study aimed to identify potential anti-inflammatory bioactive peptides from oligopeptide in white tea using network pharmacological and molecular docking approaches， validate their in vitro anti-inflammatory activity， and elucidate their underlying mechanisms and pathways， so as to provide a theoretical foundation for the high-value development and utilization of white tea.【Method】 A white tea oligopeptide database was established using ultra-high performance liquid chromatography-mass spectrometry （UPLC-MS/MS）， and otential bioactive peptide targets were identified via the SEA Search Server database. Inflammation-related targets were retrieved from the GeneCards， OMIM， and DisGeNET databases. A protein-protein interaction （PPI） network was established to screen core targets. KEGG signaling pathway enrichment analysis of key targets was performed using the Metascape database. Molecular docking validated was using AutoDock 4.2.6. For in vitro validation， a lipopolysaccharide was used to induce the RAW264.7 cellular inflammation model for determinating the releases of nitric oxide （NO）， tumor necrosis factor-α （TNF-α）， and interleukin-1β （IL-1β）. Western blotting was used to analyze the phosphorylation levels of key proteins in the PI3K/Akt/NF-κB signaling pathway.【Result】 A total of 350 potential bioactive peptide targets and 15404 inflammation-related targets were identified， with 317 potential overlapping targets of bioactive peptides and inflammation-related diseases. Through network pharmacology screening， four key anti-inflammatory peptides （LFFPR， EECFGC， SDSETRFLR， and LFHDLPPN） and three core targets （MMP9， IL-1β， and SRC） were identified. Molecular docking revealed that LFFPR， EECFGC， SDSETRFLR， and LFHDLPPN had average binding energies of -7.3， -6.4， -7.8， and -7.7 kcal/mol， respectively， with the core targets， indicating strong binding affinities. KEGG signaling pathway enrichment analysis showed that the PI3K-Akt signaling pathway and the NF-κB signaling pathway were potential anti-inflammatory pathways. In vitro experiments showed that all four peptides significantly inhibited the release of nitric oxide （NO） and inflammatory factors （TNF-α， IL-1β） （P<0.05， the same below）. Western blotting revealed that SDSETRFLR and LFHDLPPN significantly down-regulated protein expressions in the PI3K/Akt/NF-κB signaling pathway.【Conclusion】 Based on network pharmacological and molecular docking technologies， four key anti-inflammatory peptides are identified from white tea. Among them， SDSETRFLR and LFHDLPPN exert their anti-inflammatory effects through a dual mechanism： directly inhibiting the secretion of inflammatory mediators and cytokines， and suppressing the inflammatory cascade reaction by blocking the expression of PI3K/Akt1/NF-κB signaling pathway. Anti-inflammatory peptides in white tea is potential in preventing inflammation.