Unveiling the vitro toxic impact of nanosized copper on PC12 cells: A Ferroptosis-driven mechanism

Latest findings indicate that minute copper particles, specifically nano-Cu with a diameter falling between 15 and 30 nm, may well be capable of triggering neurotoxic effects. In an effort to delve into the cytotoxic impacts of nano-Cu, we subjected PC12 cells (the cell line commonly employed as a surrogate for neuron studies in a laboratory setting) to different dosages (ranging from 0 to 50 μg/mL) of this nanomaterial. Cell viability assay was performed using the Cell Counting Kit-8 (CCK-8). The oxidative stress levels induced by nano-Cu in PC12 cells were assessed by measuring reactive oxygen species (ROS) generation, malondialdehyde (MDA) levels, superoxide dismutase (SOD) activity, and mitochondrial membrane potential (JC-1). Additionally, potential toxicity mechanisms were screened by assessing ferroptosis through measuring glutathione (GSH) levels and the expression of GPX4, ACSL4, FPN-1, TFR1, HO-1, Nrf2, COX2, and FTH1. Results showed that nano-Cu exposure reduced cell viability and antioxidant capacity in a concentration-dependent manner, accompanied by elevated oxidative stress, lipid peroxidation, and mitochondrial dysfunction. Notably, the expression levels of ferroptosis-related markers were significantly altered, indicating the involvement of ferroptosis in nano-Cu-induced cytotoxicity. These findings provide critical insights into the role of ferroptosis in nano-Cu-mediated neurotoxicity, offering a deeper understanding of its underlying mechanisms.