Prenatal Indomethacin Exposure Is Associated With Autism‐Relevant Behavioral Alterations Linked to Oxidative Stress and Altered Autophagy‐Related Signaling

Autism spectrum disorder (ASD) arises from interactions between genetic predisposition and prenatal environmental insults. Prostaglandin E 2 (PGE 2 ), derived from cyclooxygenase‐2 (COX‐2), regulates neuronal differentiation and glia–neuron signaling; thus, its inhibition during gestation may impair neurodevelopment. To examine whether prenatal exposure to indomethacin, a non‐selective COX inhibitor, induces autism‐like alterations in rats with possible sex differences. Sixteen pregnant Wistar rats received indomethacin (1 mg/kg, gavage) on Gestational Days 10–14. Adult offspring (P50–P54) underwent behavioral tests (open‐field, rearing, rotarod, and three‐chamber sociability). Hippocampal and cerebellar tissues were analyzed for PGE 2 , LC3B (ELISA), MDA (TBARS), neuronal density, and GFAP expression (CA1, CA3 – Nissl, GFAP staining). Indomethacin reduced sociability, exploration, and motor performance, especially in males, and decreased PGE 2 and LC3B while increasing MDA ( p < 0.001). Histologically, CA1 neuronal counts increased while GFAP immunoreactivity was markedly elevated in hippocampal (CA1 and CA3) and cerebellar regions ( p < 0.05), reflecting astroglial activation and neuroinflammatory remodeling. Prenatal inhibition of prostaglandin synthesis may disturb PGE 2 signaling, leading to oxidative stress, altered autophagy‐related signaling, and glial activation, which together contribute to ASD‐relevant behavioral outcomes.