Perfluorodecanoic acid (PFDA) induces oxidative stress and autophagy dysregulation in HGrC1 cells.

Perfluorodecanoic acid (PFDA), a long-chain per- and polyfluoroalkyl substance (PFAS), is an emerging environmental toxicant, though little is known on its impact on female reproduction. This study investigates the cellular effects of PFDA on a human granulosa cell line, HGrC1, focusing on oxidative stress and autophagy pathways. HGrC1 cells were exposed to increasing concentrations (0.01 - 10 µM) of PFDA for 24, 48, 72, and 96 h. Notably, 10 µM PFDA for 48 h resulted in approximately 50 % reduction in cell viability, activation of cleaved caspase-3, reduced cell density, and altered morphology characterized by spindle-shaped elongation. qRT-PCR revealed significant downregulation of key antioxidant genes, particularly catalase. PFDA exposure markedly increased intracellular reactive oxygen species levels. Western blot analysis of the p62-Keap1-Nrf2 signaling pathway showed decreased KEAP1 protein levels and strong nuclear accumulation of NRF2 at 10 µM PFDA, supported by elevated expression of the downstream target HO-1. In parallel, PFDA disrupted autophagy regulation. Accumulation of p62, along with increased levels of LC3A/B, suggested impaired autophagic flux. Together, these findings demonstrate that PFDA compromises granulosa cell survival by inducing oxidative stress, altering antioxidant gene expression, and dysregulating autophagy. Given the central role of granulosa cells in follicular development and hormone synthesis, PFDA-induced toxicity may have significant implications for ovarian function and female fertility.