Changyong Lee, Rasha A Barakat, Kate E Taylor, Kendra L Clark
Per- and polyfluoroalkyl substances (PFAS) are persistent environmental contaminants increasingly linked to female reproductive toxicity. Among them, perfluorodecanoic acid (PFDA) has been implicated in ovarian dysfunction, yet the molecular basis of its ovotoxicity remains insufficiently defined. Here, we investigated PFDA-induced granulosa cell injury and examined whether kaempferol, a naturally occurring flavonoid, could mitigate these effects using an integrative approach combining network toxicology, single-cell transcriptomic projection, and in vitro validation. Computational analysis identified 121 shared targets among PFDA, kaempferol, and ovotoxicity-related gene sets, with enrichment in oxidative stress, inflammatory, and nuclear receptor-associated pathways. Single-cell projection of PFDA- and kaempferol-associated gene sets onto a human ovarian reference atlas localized these signatures across multiple ovarian cell populations, including granulosa cell clusters. In the human granulosa cell line HGrC1, PFDA significantly reduced cell viability, increased reactive oxygen species (ROS), impaired nuclear factor erythroid 2-related factor 2 (NRF2)-associated antioxidant defense, shifted apoptosis-related gene expression toward a pro-apoptotic profile, and suppressed steroidogenic gene expression. Co-treatment with kaempferol alleviated several of these changes, lowering ROS levels and partially restoring antioxidant and steroidogenic markers. Together, these findings indicate that PFDA disrupts granulosa cell homeostasis through redox imbalance and modulation of steroidogenesis-related gene expression, while kaempferol mitigates part of this response in the present in vitro model. This study provides mechanistic insight into PFDA-induced granulosa cell toxicity and supports further investigation of bioactive flavonoids as modulators of pollutant-related ovarian injury.