DEVELOPMENT OF AN IN VITRO FERROPTOSIS MODEL FOR EVALUATING NEUROPROTECTIVE AGENTS

Resumo

INTRODUCTION: The increase in global life expectancy is a significant achievement, yet itis accompanied by a higher incidence of age-related diseases such as Alzheimer’s andParkinson’s. Among the mechanisms implicated in these conditions, ferroptosis—a form ofregulated cell death dependent on iron and oxidative stress—has gained prominence in thepast decade. This pathway has been associated with the pathophysiology of severalneurodegenerative disorders. RSL-3, a known inhibitor of glutathione peroxidase 4 (GPX4),is commonly used to induce ferroptosis in cell-based models. OBJECTIVE: This study aimedto establish a ferroptosis model using differentiated SH-SY5Y human neuroblastoma cellstreated with RSL-3 and to assess the protective potential of deferoxamine, an iron chelator.MATERIALS AND METHODS: SH-SY5Y cells were plated, differentiated, and treated withincreasing concentrations of RSL-3 (0.25–25 µM). Deferoxamine was then tested atconcentrations of 5 to 100 µM in co-treatment with RSL-3 at 10 and 25 µM. After 24 hours,cell viability was evaluated using the MTT and Neutral Red (NR) assays. Results wereexpressed as percentages relative to the control group (0.625% DMSO) and analyzed usingone-way ANOVA followed by Tukey’s post hoc test, with significance set at p<0.05.RESULTS: RSL-3 significantly reduced cell viability at 5, 10, and 25 µM in the MTT assay,and at 10 and 25 µM in the NR assay. Deferoxamine conferred protection only at lowerconcentrations, suggesting that iron accumulation is a key driver of RSL-3-inducedferroptosis, while higher doses of the chelator may have limited efficacy. CONCLUSION:RSL-3 effectively induces ferroptosis in differentiated SH-SY5Y cells, especially at 10 µM.This model may contribute to the study of neurodegenerative disease mechanisms and thedevelopment of potential neuroprotective strategies.