Maternal choline and prebiotic supplementation ameliorate PCOS traits and reduce intergenerational transmission in rats.

Polycystic ovary syndrome (PCOS) is a common endocrine disorder with strong heritability, driven by complex metabolic, epigenetic, and gut-microbial dysregulation. While both epigenetic and microbial axes are implicated in PCOS, their integration and contribution to intergenerational transmission remain poorly understood. We investigated whether maternal choline (methyl-donor) and galactooligosaccharide (GOS, prebiotic) supplementation could restore gut-epigenome homeostasis and prevent vertical transmission of PCOS traits in rats. PCOS was induced using dehydroepiandrosterone plus high-fat diet (DHFD). Reproductive, metabolic, epigenetic, and microbial outcomes were assessed in F0 females and F1 offspring. DHFD rats displayed hyperandrogenism, cystic ovaries, insulin resistance, and gut dysbiosis. Choline and GOS restored these phenotypes, normalized epigenetic enzymes (DNMT1, DNMT3a, HDAC3), histone modifications (H3K9ac, H3K9me2), and gene-specific methylation patterns. Beneficial bacteria positively correlated with hypomethylated reproductive/metabolic genes, while pathogenic taxa correlated with hypermethylated genes, with folic acid emerging as a potential biochemical bridge. F1 female offspring of DHFD rats inherited aberrant methylation and dysbiosis in a sex-specific manner, maternal supplementation prevented this transmission. Fertility outcomes, including conception efficiency and pregnancy success, were markedly improved, with choline showing strongest effects. Maternal choline and GOS supplementation restore gut-epigenome homeostasis, ameliorate PCOS phenotypes, and reduce intergenerational transmission of disease traits.