The NEUROPROTECTIVE POTENTIAL OF IMEGLIMIN AGAINST HYDROGEN PEROXIDE-INDUCED OXIDATIVE STRESS IN SH-SY5Y CELLS: AN IN SILICO AND IN VITRO STUDY

Objectives: Oxidative stress plays a central role in neuronal injury and neurodegenerative disorders. The present study aimed to evaluate the protective effect of imeglimin against hydrogen peroxide (H2O2)-induced oxidative stress in SH-SY5Y human neuroblastoma cells and to explore its potential molecular interactions with oxidative stress-related protein targets using in silico approaches. Methods: In vitro experiments were performed using SH-SY5Y cells exposed to H2O2-induced oxidative stress. Cell viability was assessed using the MTT assay, lipid peroxidation was evaluated by measuring malondialdehyde levels, and apoptotic cell death was analysed using acridine orange/ ethidium bromide staining. In silico molecular docking was carried out to assess the interaction of imeglimin with amyloid beta, brain-derived neurotrophic factor, forkhead box protein 3, and Kelch-like ECH-associated protein 1 (KEAP1). Molecular dynamics (MD) simulation and MM/GBSA binding-free energy calculations were performed for the imeglimin-KEAP1 complex to evaluate interaction stability. Results: Imeglimin did not exhibit cytotoxic effects in SH-SY5Y cells across the tested concentration range. H2O2 exposure resulted in a dose-dependent reduction in cell viability and increased lipid peroxidation. Co-treatment with imeglimin improved cell viability, reduced lipid peroxidation, and attenuated apoptotic and necrotic cell death under oxidative stress conditions. Molecular docking analysis demonstrated favorable binding of imeglimin with all selected protein targets, with the highest binding affinity observed for KEAP1. MDs simulation indicated stable interaction of imeglimin with KEAP1 throughout the simulation period. Conclusion: The findings suggest that imeglimin exerts cytoprotective effects against H₂O₂-induced oxidative stress in SH-SY5Y neuronal cells and exhibits favorable molecular interactions with oxidative stress-related protein targets. These results provide preliminary evidence supporting the potential neuroprotective role of imeglimin and warrant further mechanistic and in vivo investigations.