PFOA induces apoptosis in primary cortical neurons via GLS2 inhibition-mediated ROS accumulation

Perfluorooctanoic acid (PFOA) is a persistent environmental pollutant whose bioaccumulative nature and potential neurotoxicity have raised increasing health concerns. However, its adverse effects on cortical neurons and the underlying mechanisms remain poorly understood. In this study, primary cortical neurons isolated from mice were exposed to different concentrations of PFOA (0, 25, 50, 100, 200, and 400 μM) for 48 h. Intracellular reactive oxygen species (ROS) levels, cell viability, and apoptosis were evaluated, while neuronal morphology, NF-κB activation, and nuclear translocation were examined by immunofluorescence. Western blotting was used to assess the expression of NF-κB, p-NF-κB, GLS2, Bax, Bcl-2, Caspase-3 and Cleaved Caspase-3. PFOA exposure markedly shortened neurite branch length, reduced branch number, and decreased neuronal soma size. Increasing PFOA concentrations induced a dose-dependent accumulation of ROS, enhanced NF-κB activation and nuclear translocation, and elevated apoptosis rates. Transcriptomic analysis using the GEO dataset GSE254408 revealed significant downregulation of GLS2, a gene involved in glutamate metabolism and neuronal homeostasis. Glutamate assay revealed that PFOA treatment (100, 200, and 400 μM) led to a significant reduction in intracellular glutamate levels. Consistently, Western blotting showed that in the high-dose PFOA groups (200 μM and 400 μM), GLS2 and Bcl-2 expression decreased, whereas Bax, NF-κB, p-NF-κB, Caspase-3, and Cleaved Caspase-3 were upregulated. Notably, GLS2 overexpression partially attenuated PFOA-induced ROS accumulation and neuronal apoptosis. Collectively, these findings suggest that PFOA exposure may promote neuronal apoptosis through suppression of GLS2 and subsequent ROS accumulation. This study highlights GLS2 as a potential molecular target in PFOA-induced neurotoxicity and provides mechanistic insights relevant to hazard identification and future therapeutic strategies.