Endocrine Disruptors, Short-Chain Per- And Polyfluoroalkyl Substances, Promote Aldosterone Secretion in Human Adrenocortical Cells

Objective: Per- and polyfluoroalkyl substances (PFAS) are synthetic chemicals widely used in industrial and consumer products for their stability and water- and oil-repellent properties. Our previous study demonstrated that long-chain PFAS, such as perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS), significantly stimulate aldosterone biosynthesis via oxidative stress mechanisms in human adrenocortical cells. Due to their known toxicity and bioaccumulation, long-chain PFAS act as endocrine disruptors and have been increasingly restricted in Europe. As replacements, short-chain PFAS, including perfluorohexanoic acid (PFHxA) and perfluoroether carboxylic acid (C6O4) as substitutes for PFOA, and perfluorobutanesulfonic acid (PFBS) and Perfluoro-4-ethylcyclohexanesulfonate (PFECHS) as substitutes for PFOS, have gained widespread use despite limited toxicological data and unproven safety. Potassium 9-chlorohexadecafluoro-3-oxanonane-1-sulfonate (F-53B), instead, is used as a substitute for both PFOA and PFOS. In this study, we evaluated the effects of short-chain PFAS on aldosterone production in HAC15 human adrenocortical cells. Design and method: We exposed HAC15 cells to short-chain PFAS (PFHxA, C6O4, PFBS, PFECHS, F-53B) at 1, 10, and 100 ng/mL for 24 hours, consistent with human exposure levels. Cell viability was assessed using the Incucyte® SX5 Instrument. Aldosterone synthase (CYP11B2) gene expression and aldosterone levels were quantified by real-time RT-PCR and ELISA, respectively. Results: We found that PFHxA, C6O4, PFBS, PFECHS, and F-53B did not affect HAC15 cell viability. However, they significantly increased CYP11B2 gene expression and aldosterone secretion. Notably, these increases were comparable to those observed with long-chain PFAS. Conclusions: Our findings demonstrate that short-chain PFAS increase aldosterone secretion and suggest that they may act as endocrine disruptors in the adrenal gland. Given the well-established role of aldosterone in blood pressure regulation and cardiovascular disease, these data raise serious safety concerns about short-chain PFAS and underscore the need for further toxicological and mechanistic studies to evaluate their health impact.