Autism spectrum disorder (ASD) affects approximately 1% of children worldwide, with its prevalence continuing to rise. Prenatal exposure to valproic acid (VPA) has been reported to increase the risk of ASD in offspring to around 4%, leading to restrictions on its use during pregnancy. However, the molecular mechanisms by which VPA contributes to ASD remain unclear. In this study, human neuronal cells (SH-SY5Y) were treated with varying concentrations of VPA to determine the optimal dose for analysis. RNA sequencing performed at this concentration revealed 62 upregulated and 144 downregulated genes. Functional analysis of the most significantly altered genes identified HTR6, PTGS1, and LRP2 as potential contributors to ASD, given their roles in synapse formation, neurotransmitter regulation, and receptor signaling during neurodevelopment. Network analysis showed that PTGS1 exhibited extensive interactions with multiple genes, forming a strong regulatory hub, whereas LRP2 and HTR6 did not display significant network connectivity. These findings highlight transcriptional changes induced by VPA exposure and suggest key genes that may serve as targets for therapeutic strategies aimed at modulating gene function in the prevention or treatment of ASD.