Neuroprotective efficacy of hentriacontane against rotenone-induced apoptosis in SH-SY5Y cells: In silico and in vitro evidence of GSK3β association.

Neurodegenerative diseases, such as Alzheimer's disease (AD), are marked by gradual neuronal loss, frequently associated with mitochondrial dysfunction. Hentriacontane is a naturally occurring long-chain saturated hydrocarbon found in beeswax and certain herbal plants, and has antioxidant, anti-inflammatory and cytoprotective properties, suggesting its therapeutic potential in oxidative stress-associated neurodegeneration. Rotenone, a mitochondrial complex I inhibitor, induces oxidative stress and apoptosis, making it a widely used neurotoxic agent for investigating neurodegenerative disorders like AD. We initially performed in silico molecular docking, molecular dynamics, MM/GBSA, and ADMET pharmacokinetic analyses of hentriacontane against GSK3β to evaluate binding affinities and energies and assess the stability and dynamics of protein-ligand complexes. An in vitro drug safety assay was conducted using the MTT assay in SH-SY5Y cells. The cells were pretreated with 100 μM rotenone two hours before the assay. A 96-well tissue culture-grade microplate was prepared with 100 μL of cell suspension (1 × 10