Luteolin alleviates inflammation by inhibiting the PI3K-Akt-FOXO3a pathway

Luteolin is a flavonoid commonly found in various plants and has been shown to possess a wide range of biological activities through the modulation of multiple signaling pathways. Despite its well-documented effects, the underlying molecular mechanism responsible for its anti-inflammatory properties remains unclear. Network pharmacology suggested that luteolin may exert anti-inflammatory effects by modulating the PI3K/Akt/FOXO3a signaling cascade. Consistent with this prediction, in an LPS-stimulated RAW264.7 macrophage model, luteolin significantly decreased the phosphorylation of PI3K, Akt, and FOXO3a, while also lowering the ratios of p-PI3K/PI3K, p-Akt/Akt, and p-FOXO3a/FOXO3a. Additionally, Molecular docking combined with molecular dynamics simulations indicated that luteolin can bind to PI3K, Akt, and FOXO3a to form stable protein-ligand complexes, supporting the presence of strong binding affinity/interactions between luteolin and these targets. Further validation using cellular thermal shift assay, isothermal dose-response, and drug affinity response target stability techniques confirmed that luteolin directly interacts with PI3K. These results provide compelling evidence supporting luteolin's potential clinical use, enhance our understanding of the mechanisms driving inflammation, and lay the groundwork for the development of novel anti-inflammatory therapies inhibiting the PI3K-Akt-FOXO3a pathway.