Objective Stroke is a leading cause of death and disability worldwide, with ischemic stroke accounting for 80%-85% of cases. Despite the prevalence, effective treatments remain scarce. The compelling evidence suggest that high concentrations of ATP in the brain post-stroke can trigger irreversible neuronal damage and necrosis, contributing to a range of neurocellular dysfunctions. Pyroptosis, a recently identified form of programmed cell death, is characterized by caspase-1 activation and the action of the Gasdermin D (GSDMD) protein family, leading to cell perforation and inflammatory death.Methods In this study, human neuroblastoma SH-SY5Y cells were used to investigate the mechanisms of ATP-induced neurotoxicity and the protective effects of hydrogen sulfide (H₂S) against this toxicity through the antagonization of pyroptosis. We employed CCK-8 and LDH assays to assess cell viability. YO-PRO-1 fluorescent dyes and flow cytometry were conducted for detecting changes in cell membrane permeability. Western blot analysis was used to measure protein levels associated with cellular dysfunction.Results Our results indicate that high concentrations of ATP enhance cytotoxicity and increase cell membrane permeability in SH-SY5Y cells, that are mitigated by the H₂S donor NaHS. Furthermore, ATP was found to promote the activation of the NOD-like receptor pyrin domain-containing 1 (NLRP-1), caspase-1, and the cleavage of GSDMD, with NaHS significantly attenuating these effects.Conclusion Our research suggests that H₂S protects SH-SY5Y cells from ATP-induced neurotoxicity through a mechanism mediated by the NLRP1, caspase-1, and GSDMD pathway.