Exploring the antidepressant-like effect of stigmasterol using network pharmacology with molecular docking and in vivo experimental validation.

With a global prevalence of 3.8% and affecting approximately 300 million people, depression remains a global burden calling for renewed efforts including novel antidepressants to address treatment gaps. Stigmasterol, an unsaturated naturally abundant phytosterol with reported neuropsychiatric activity in preclinical studies, was studied for potential antidepressant-like activity and mechanism of action using computational and experimental approaches. Using network pharmacology, we identified potential key targets of major depressive disorder and stigmasterol (STG) by analysing intersection genes for protein-protein interaction, Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses. We subsequently carried out molecular docking of stigmasterol with key targets identified to confirm antidepressant activity and potential mechanism(s) while the forced swim test (FST) and tail suspension test (TST) were used for experimental validation. To experimentally validate the involvement of monoaminergic mechanism(s) in STG's action, mice were pretreated with selective inhibitors of monoamine synthesis and storage after which the antidepressant-like effects of STG was re-evaluated in FST. Forty intersection target genes were obtained with AKT1, TP53 and IL1B as well as MAO