Baicalin alleviates repeated superovulation induced ovarian dysfunction and oocyte impairment by suppressing excessive glycolysis

Repeated superovulation is widely applied in livestock reproduction programs to enhance ovulation rate and embryo yield, but increasing evidence indicates that intensive stimulation may disrupt ovarian homeostasis and impair oocyte quality. This study evaluated whether baicalin, a bioactive flavonoid, can mitigate ovarian dysfunction induced by repeated superovulation. Female mice were subjected to four cycles of superovulation with or without baicalin treatment. Physiological parameters and reproductive outcomes were recorded. Ovarian and fecal metabolomic profiles, as well as gut microbiota composition, were analyzed by LC–MS and 16S rRNA sequencing. Histological and molecular analyses were performed to assess ovarian morphology, steroidogenesis, AMPK/Foxo3a-mediated glycolysis, and oocyte quality. Repeated superovulation induced follicular atresia, metabolic disturbances characterized by excessive glycolysis, suppression of AMPK/Foxo3a signaling, gut microbial dysbiosis, and reduced MII oocyte quality. Baicalin markedly alleviated these alterations by restoring metabolic and signaling homeostasis, improving microbial balance, and preserving oocyte maturation and spindle integrity. Collectively, these findings provide new insights into the metabolic basis of ovarian dysfunction induced by repeated superovulation and suggest that baicalin may help preserve ovarian function and oocyte quality under conditions of intensive ovarian stimulation, with potential relevance to the sustainability of donor animals in livestock reproduction programs.