Orexin A Ameliorates Ovarian Dysfunction in Premature Ovarian Insufficiency Mice by Inhibiting Granulosa Cell Pyroptosis via the AMPK/NLRP3 Pathway

ABSTRACT Premature ovarian insufficiency (POI) is a condition characterized by the early depletion of ovarian follicles, leading to infertility and various systemic complications. Granulosa cell (GC) pyroptosis contributes significantly to the pathogenesis of POI. Orexin A, a neuropeptide involved in regulating wakefulness, has been shown to exert anti‐inflammatory effects. This study investigates the potential protective role of orexin A in POI by targeting pyroptosis in ovarian GCs. We found that orexin A significantly reduced oxidative stress and the activation of the NLRP3 inflammasome in POI mice, thereby improving serum hormone levels and follicle count. Additionally, orexin A inhibited pyroptosis in cyclophosphamide (CTX)‐treated KGN cells by downregulating NLRP3, caspase‐1, and gasdermin D (GSDMD) expression. These effects were mediated through the activation of adenosine 5'‐monophosphate‐activated protein kinase (AMPK) signaling, which is known to regulate cellular metabolism and suppress inflammasome activation. In conclusion, orexin A has the potential to alleviate POI by inhibiting NOD‐like receptor thermal protein domain‐associated protein 3 (NLRP3) inflammasome‐mediated pyroptosis and activating AMPK signaling, offering a promising therapeutic approach for POI treatment.