Zerumbone-mediated post-ischemic neuroprotection: Reduction of ferroptosis through TFR1 downregulation in vitro

Emerging studies have identified ferroptosis as a promising therapeutic target, the inhibition of which is hypothesized to mitigate brain injury and subsequent neuronal death following stroke. Zerumbone, a phytochemical sesquiterpene isolated from Zingiber zerumbet Smith, exhibit diverse therapeutic properties across a range of neurological disorders. This study aimed to elucidate the postischemic neuroprotective effects and regulatory impact of zerumbone on ferroptosis-mediated cell death following oxygen‒glucose deprivation/reperfusion (OGD/R) injury. We employed an in vitro OGD/R SH-SY5Y cell model of stroke to evaluate the postischemic neuroprotective effects of zerumbone, a lead molecule identified through literature studies. Moreover, assays were performed to assess how zerumbone affects lipid peroxide levels, intracellular reactive oxygen species (ROS), and mitochondrial membrane integrity. Furthermore, molecular docking simulations were carried out to determine the targets, and western blotting was performed to examine TFR1 protein expression. Zerumbone (0.5 µM) treatment at 1-hour postischemia increased cell viability (72.11 ± 0.98) and mitigated OGD/R-induced ischemic injury. Zerumbone significantly decreased intracellular ROS levels and lipid peroxide production while increasing mitochondrial membrane integrity, suggesting that zerumbone ameliorated OGD/R-induced ischemic injury by inhibiting ferroptosis in vitro. This finding was corroborated by our western blot analysis, which revealed that the antiferroptotic role of zerumbone was distinctly mediated through the downregulation of transferrin receptor 1 (TFR1) protein expression. This communication, for the first time, highlights the feasibility of zerumbone as a promising adjunctive neuroprotective agent against ferroptosis cell death in the context of cerebral stroke. This study lays the groundwork for subsequent in-depth investigations to fully elucidate its therapeutic potential in ischemic stroke treatment.