Deciphering the Antidepressant Potential of Piper betle L. Essential Oil and Its Inclusion Complex: Network Pharmacology and Experimental Insights.

This study evaluates the antidepressant effects of Piper betle L. leaf essential oil (PBEO) and its inclusion complex (PBECD) in mice whilst exploring potential mechanisms. Gas chromatography-mass spectroscopy (GC-MS) identified PBEO's compounds, and β-cyclodextrins were used to enhance stability and bioavailability by forming PBECD. Characterization was performed using various analytical techniques. ADME pharmacokinetics, network pharmacology, and molecular docking predicted active compounds, antidepressant targets, and signalling pathways. The Chronic Unpredictable Mild Stress (CUMS) model was used to validate findings, with fluoxetine (20 mg/kg) as the standard. Mice received PBEO and PBECD at 50 mg/kg and 100 mg/kg doses. Behavioural tests, including the open field, sucrose preference, Y-maze, and forced swim tests, assessed antidepressant effects. Toxicity was evaluated during dosing. Serum levels of IL-1β and TNF-α were measured using ELISA, whilst biochemical markers such as glutathione (GSH) and malondialdehyde (MDA) were analysed. ADME and network pharmacology identified six active components targeting 244 depression-related proteins, primarily in the MAPK signalling pathway. Molecular docking confirmed strong binding affinities between the ligands and the obtained proteins from network pharmacology. In vivo results showed PBEO and PBECD reduced depressive behaviours, with PBECD demonstrating superior effects. PBECD (100 mg/kg) significantly reduced immobility time by 42.5% in the forced swim test, increased sucrose preference by 36.8%, and improved Y-maze alternation by 31.7% compared to the control group. Serum levels of IL-1β and TNF-α decreased by 48.3% and 41.2% respectively, whilst GSH levels rose by 35.6% and MDA levels dropped by 39.4%, indicating reduced oxidative stress and inflammation. Modulating MAPK signalling, PBECD emerges as a promising candidate for antidepressant activity than PBEO in preclinical findings, as its complexation with cyclodextrin improves the oils' stability, solubility, and bioavailability, resulting in enhanced efficacy.