Abstract
Autism spectrum disorder (ASD) is a pervasive neurodevelopmental disorder characterized by impaired social interactions and communication, repetitive, or stereotyped behavior. Docosahexaenoic acid (DHA), an essential polyunsaturated fatty acid, has been demonstrated to exert anti-oxidative stress and anti-inflammatory properties, while also promoting myelin development and neural differentiation and development. However, it remains uncertain whether DHA can ameliorate autistic-like behaviors, and if so, the underlying mechanisms are still unclear. Here, we established a neonatal maternal separation (NMS) rat model and treated it with DHA (80 mg/kg/day, i.p.). The results showed DHA treatment significantly alleviated autism-like behaviors in the NMS rats during their juvenile period. Subsequently, we employed network pharmacology analysis and molecular docking methods to screen potential targets of DHA in ASD therapy. Through Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Wikipedia enrichment analysis, we identified anti-oxidative stress, anti-inflammatory and JNK signaling pathway that might be associated with DHA-mediated improvement of autistic-like behaviors. Furthermore, western blotting assays showed DHA significantly downregulated the expression levels of p-JNK and c-JUN, while upregulating the expression levels of NRF2, HO-1, SOD1, and CAT. In addition, enzyme-linked immunosorbent assay results showed DHA effectively reduced the production of pro-inflammatory factors, such as TNF-α, IL-6, and IL-1β. Collectively, our study predicted and validated that DHA exhibits the potential to improve autistic-like behaviors induced by NMS in rats by suppressing JNK activation and inhibiting oxidative stress and inflammatory response. These findings suggest that DHA may be a potential therapeutic agent for the treatment of ASD.