Gestational exposure to the artificial sweetener erythritol reprograms ovarian function through AMH suppression and oxidative stress-mediated disruption of autophagy and PI3K signaling.

Abstract

Erythritol is a widely used non-nutritive sweetener generally regarded as safe. However, recent emerging evidence has begun to reveal potential adverse effects, including possible associations with cancer and cardiovascular disease. Nevertheless, its potential impact on female reproductive health remains largely unexplored. The present study investigated the transgenerational effects of gestational erythritol exposure on ovarian morphology and molecular signaling in Wistar rats. Pregnant females were administered 0.4 or 4 g/kg/day of erythritol from gestational day 6 until parturition. Ovarian histopathological architecture along with oxidative stress and molecular signaling markers were evaluated in F1 and F2 female offspring at the prepubertal stage. We found that in utero erythritol exposure markedly disrupted folliculogenesis, increasing primordial follicle counts while significantly reducing Graafian follicles. Histological examination revealed notable anomalies, including degenerating and multi-oocyte follicles and granulosa cell pyknosis. Oxidative stress was significantly elevated across generations, as evidenced by increased malondialdehyde (MDA) levels and reduced superoxide dismutase (SOD) and glutathione (GSH) activity. Anti-Müllerian hormone (AMH) was significantly diminished, indicative of impaired granulosa cell functionality and reduced follicular competence. Autophagy markers, LC3 and Atg5, were markedly suppressed in F2 offspring, strongly implicating a transgenerational susceptibility mechanism potentially attributable to epigenetic reprogramming, given that global DNA methylation levels were significantly reduced in F2 offspring. Furthermore, elevated erythritol levels in F2 offspring were associated with reduced PI3K-p85 expression, indicating compromised follicular survival signaling. Collectively, these findings demonstrate that gestational erythritol exposure promotes oxidative stress, disrupts autophagy, impairs steroidogenesis, and inhibits PI3K signaling, culminating in transgenerational ovarian dysfunction.