Exploring the therapeutic potential of the ayurvedic formulation Vilwadi Gulika through an integrated approach employing network pharmacology and microbiome analysis

Abstract

Vilwadi Gulika is a classical Ayurvedic polyherbal formulation traditionally used for snake bites, insect bites, gastrointestinal disorders, toxicological conditions, and inflammatory states. Despite its long use, systematic scientific evaluation of its molecular mechanisms and microbial characteristics remains limited. In this study, the therapeutic potential of Vilwadi Gulika was investigated using an integrated approach combining network pharmacology and microbiome analysis. Network pharmacology analysis was performed using phytochemicals derived from 13 medicinal plants, comprising of 3004 phytochemicals, curated based on the traditionally prescribed plant parts. Phytochemical data were retrieved from 3 databases (IMPPAT, Dr. Duke’s Phytochemical and Ethnobotanical Database, and KNApSAck), followed by molecular target prediction using Binding DB and Swiss Target Prediction accumulating 68,635 targets. 1,347 Disease and 238 pathway annotation led to the construction of interaction networks with 1,047 genes. 6 gastrointestinal diseases were identified yielding 7 key genes, TNF, PTGS2, JAK2, NOS2, SRC, TGFB1, and CXCL8. Colitis-related targets (NOS2, SRC, PTGS2) were associated with ellagic acid, eugenol, elemicin, and ethyl gallate. Crohn’s disease–associated JAK2 showed interactions with multiple terpenoids and alkaloids, while TGFB1 and TNF were linked to phytochemicals relevant to duodenal ulcer, ileocolitis, and regional enteritis. Microbiome profiling was carried out across different stages of Vilwadi Gulika preparation, including in-house, Vaidya-prepared, and commercial formulations, using 16S rRNA gene sequencing. The in-house formulation was dominated by Firmicutes (97.37%), with Aerococcus (94.15%) and Aerococcus viridans (71.65%) as the predominant genus and species, respectively. Overall, the findings indicate that Vilwadi Gulika exhibits a layered therapeutic profile involving inflammatory gene modulation and a structured microbial community, supporting its traditional use in gastrointestinal disorders.