Neuroprotective and Antioxidant Effectsof Benincasa hispida Ethanolic Extract inD-Galactose-induced SH-SY5Y Cells

Abstract

Aim: The present study aimed to evaluate the neuroprotective and antioxidant potential of Benincasa hispida
ethanolic extract (BHE) against d-galactose-induced neurotoxicity in human neuroblastoma SH-SY5Y cells,
an in vitro model relevant to Alzheimer’s disease. Materials and Methods: SH-SY5Y cells were cultured and
differentiated using retinoic acid, followed by induction of neurotoxicity with d-galactose (300 μM). Cells were
pretreated with different concentrations of BHE. Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-
2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) assays. Oxidative stress markers and
antioxidant enzymes, including reactive oxygen species, malondialdehyde assay, reduced glutathione (GSH),
superoxide dismutase, catalase, Glutathione peroxidase (GPx), and neurotransmitter-related enzymes, were estimated
using enzyme-linked immunosorbent assay-based methods. Results: D-galactose significantly reduced cell viability,
increased LDH leakage, and elevated oxidative stress markers. Pretreatment with BHE dose-dependently improved
cell viability, reduced LDH release, restored antioxidant enzyme levels, and reduced oxidative damage. Conclusion:
BHE demonstrated significant neuroprotective and antioxidant effects against d-galactose-induced neuronal damage,
suggesting its potential as a therapeutic candidate for neurodegenerative disorders.