Comparative assessment of female reproductive toxicity from PFOA and its alternative GenX in mice and human granulosa cells

Perfluorooctanoic acid (PFOA) is a legacy perfluoroalkyl substance (PFAS) with various detrimental health effects, prompting its replacement by hexafluoropropylene oxide dimer acid (GenX). However, the female reproductive toxicity and underlying mechanism of GenX remain inadequately understood. In this study, we comparatively evaluated the impacts of PFOA and GenX on ovarian function using integrated in vivo mouse and in vitro human granulosa cell models. Our experimental findings indicated that oral exposure to PFOA significantly reduced ovarian weight, impaired follicular development, disrupted estrous cyclicity, decreased estradiol level, and induced ovarian oxidative stress and apoptosis in mice. However, GenX did not display significant toxic effects on the ovaries at the doses tested, except for reducing ovarian GPX4 expression and serum estradiol level. Notably, both PFOA and GenX at high-concentration treatment in vitro dramatically impaired the viability and proliferation, elicited ROS overproduction and mitochondrial injury, and suppressed NRF2 and HO-1 expression in cultured KGN cells. Nevertheless, at equivalent exposure doses, GenX elicited markedly decreased adverse influences on KGN cells compared to PFOA. Molecular docking simulation indicated a stronger interaction of PFOA with NRF2 than GenX. In addition, treatment with PFOA also diminished estradiol secretion and induced apoptosis in KGN cells. In summary, PFOA exposure resulted in female reproductive impairment by inducing oxidative stress and apoptosis in mouse ovaries and human granulosa cells. Although GenX exhibited comparatively low ovarian detriment relative to PFOA, its potential risk of reproductive toxicity remains a nonnegligible concern.