Maternal 3-NPA-induced oxidative stress impairs offspring ovarian function via glutamine metabolic dysregulation and NF-κB-mediated inflammation

Abstract

Oxidative stress is a well-established contributor to reproductive dysfunction, yet its intergenerational effects on offspring ovarian health remain poorly understood. This study aimed to investigate whether 3-nitropropionic acid (3-NPA)-induced maternal oxidative stress impairs ovarian development in offspring and elucidate underlying mechanisms. Female C57BL/6 mice were intraperitoneally injected 3-NPA (40 mg/kg/day) for 21 days, while the control received saline. F1 offspring were assessed for ovarian function through follicle counts, serum FSH and AMH, and fertility metrics. Untargeted metabolomics and transcriptomics were performed on ovarian tissues, complemented by immunohistochemistry for glutamine metabolism enzymes of GLS1, GS, GDH and inflammatory markers (TNF-α). F1 offspring exhibited premature ovarian insufficiency (POI)-like phenotypes, including prolonged conception intervals, reduced pregnancy rates, elevated FSH, diminished primordial follicles and AMH levels, alongside impaired fertility. Maternal 3-NPA exposure induced sustained oxidative stress. Metabolomics revealed significant dysregulation of glutamine metabolism, with compensatory upregulation of GLS1/GS/GDH enzymes. Transcriptomics identified NF-κB/TNF-α pathway activation, validated by elevated ovarian TNF-α. Maternal oxidative stress triggers persistent ovarian dysfunction in offspring through glutamine metabolic disruption and NF-κB-mediated inflammation. This study provides the first evidence that 3-NPA induces intergenerational reproductive toxicity, highlighting glutamine metabolism and inflammatory pathways as potential targets for preventing oxidative stress-associated fertility decline.