ALDH1A3-dependent ferroptosis mediates hexafluoropropylene oxide dimer acid–induced trophoblast dysfunction

M. Consistent with these in silico findings, pharmacological inhibition experiments showed that only the ferroptosis inhibitor ferrostatin-1 effectively rescued HFPO-DA-induced cytotoxicity. HFPO-DA exposure led to hallmark ferroptotic alterations, including glutathione and superoxide dismutase depletion, malondialdehyde accumulation, intracellular Fe²⁺ overload, GPX4 suppression, and increased mitochondrial superoxide production in both trophoblast cell models. Mechanistically, HFPO-DA markedly downregulated ALDH1A3 at both mRNA and protein levels. Notably, siRNA-mediated ALDH1A3 silencing alone recapitulated ferroptosis-associated biochemical and functional defects, whereas ALDH1A3 overexpression restored redox homeostasis, attenuated lipid peroxidation and iron dysregulation, and rescued trophoblast functional impairment. Collectively, these findings identify ALDH1A3 repression as a key molecular event linking HFPO-DA exposure to trophoblast ferroptosis and dysfunction, providing mechanistic insight into HFPO-DA-associated placental pathogenesis.