Exploring the anti-diabetic mechanism of selective phytochemicals identified from Gymnema sylvestre using TLC-UPLC-MS, complemented by in silico studies

Background: Diabetes is a serious metabolic condition characterized by high blood glucose levels due to inadequate insulin action or insulin resistance. It can impair various metabolic pathways in cellular metabolism, leading to significant long term effects. In the Ayurvedic system of medicine, leaves of Gymnema sylvestre (GS) have been traditionally utilized to manage diabetes. Aim of the study: This study aims is aimed to investigate the mechanism of action driven by the chemical constituents in potentially treating diabetes. Methods: Metabolite profiling of the methanolic extract from Gymnema sylvestre leaves (MLGS) was conducted using thin-layer chromatography (TLC), and ultraperformance-liquid chromatography-mass spectroscopy (UP-LCMS). Molecular Docking and MD simulations were carried out to investigate the interaction between selective phytoconstituents and the antidiabetic target, peroxisome proliferator-activated receptor (PPARγ). Results: MLGS demonstrated significant antioxidant activity, estimated using 2, 2-diphenyl-1-picrylhydrazyl (DPPH), and with an IC50 of 103.73 μg/mL. Aditionally, MLGS also exhibited substantial inhibition of α-amylase and α-glucosidase, with an IC50 of 113.49 μg/mL and 127.40 μg/mL, respectively. Using TLC-MS-bioautography, two antioxidant metabolites and three α-amylase inhibitory metabolites were identified Quercetin was identified in both α-amylase and antioxidant prominent bands. Identified phytoconstituents, namely rutin and lupeol, exhibited favorable docking interactions with PPARγ. MD simulations results indicate that rutin creates a well-built protein-ligand complex, showed minor structural modifications, and could exhibit agonistic activity with PPARγ. This suggesting the potential of rutin as a druggable compound to treat diabetes. Conclusion: These findings suggest that MLGS is enriched with antidiabetic phytoconstituents, warranting further exploration through the isolation of antidiabetic metabolites and their testing in animal models.