Summary:
A preclinical study from Chongqing Medical University and Guangzhou Women and Children’s Medical Center demonstrates that docosahexaenoic acid (DHA), a crucial omega-3 polyunsaturated fatty acid, alleviates autism-like behavioral deficits in young rats exposed to early-life maternal separation stress.
Publishing in Pediatric Discovery, researchers revealed that daily DHA supplementation restored social interaction, curtailed repetitive self-grooming, and reduced anxiety. Molecular and network pharmacology analyses showed that DHA directly targets and inhibits the pro-inflammatory c-Jun N-terminal kinase (JNK) pathway while activating NRF2 antioxidant defenses within the prefrontal cortex, offering mechanistic insights into nutritional neuroscience approaches for neurodevelopmental conditions.
Key Facts:
- Behavioral Rescue in Early Stress Models: In neonatal maternal separation (NMS) rats displaying autism-like phenotypes, daily intraperitoneal DHA (80 mg/kg) increased social interaction in three-chamber assays, reduced compulsive self-grooming, and normalized open-arm exploration in elevated plus mazes.
- Inactivation of the JNK Signaling Axis: DHA suppressed phosphorylation of JNK (p-JNK) and lowered c-JUN levels in the prefrontal cortex without altering total JNK abundance, demonstrating targeted pathway deactivation rather than protein depletion.
- Dual Anti-Inflammatory & Antioxidant Restoration: Treatment significantly reduced key neuroinflammatory cytokines (TNF-α, IL-6, IL-1β) while boosting endogenous antioxidant machinery via NRF2, HO-1, catalase (CAT), and superoxide dismutase 1 (SOD1).
Source: Children’s Hospital of Chongqing Medical University / Guangzhou Women and Children’s Medical Center
Article Body
Autism spectrum disorder (ASD) encompasses a heterogeneous continuum of early neurodevelopmental phenotypes characterized by persistent challenges in social communication, atypical sensory processing, and restricted, repetitive behavioral patterns. While the fundamental etiology involves complex polygenic and epigenetic architectures, mounting clinical evidence indicates that systemic immune dysregulation, neuroinflammation, and redox imbalances are common pathophysiological features in many autistic children.
Elevated circulating cytokines and depleted cellular antioxidant buffers create a neurotoxic cellular environment, frequently impairing neural plasticity and synaptic transmission within the prefrontal cortex (PFC), the cerebral locus governing executive control, social cognition, and repetitive impulse inhibition.
Docosahexaenoic acid (DHA), an essential long-chain omega-3 polyunsaturated fatty acid highly enriched in cerebral grey matter, is fundamental to synaptic membrane fluidity, dendritic arborization, and myelin sheath maintenance. Although dietary omega-3 interventions have shown mixed, exploratory benefits in small-scale human trials, the precise molecular mechanisms by which DHA could modify autism-related neural signaling remained elusive.
Now, a multi-method investigation published in Pediatric Discovery provides direct evidence that DHA mitigates autism-like behavioral phenotypes by dampening the hyperactive c-Jun N-terminal kinase (JNK) stress-signaling cascade and restoring antioxidant defenses in the brain.
Rescuing Sociality and Curtailing Stereotypy
The researchers, based at the Children’s Hospital of Chongqing Medical University and Guangzhou Women and Children’s Medical Center, evaluated the therapeutic potential of DHA using the neonatal maternal separation (NMS) rat model. In this established neurodevelopmental paradigm, rodent pups undergo three hours of maternal separation daily from postnatal day 1 through 21, followed by individual isolation housing until day 56—an early environmental trauma that reproduces core autism-like behaviors.
Rats administered daily DHA (80 mg/kg/day) demonstrated marked behavioral improvements between postnatal days 42 and 56:
- Social Engagement: In standardized three-chamber sociability apparatuses, DHA-treated animals spent significantly more time investigating novel conspecific stranger rats during both introductory sociability and social novelty phases.
- Repetitive Behaviors: Compulsive stereotypic self-grooming bouts—a behavioral proxy for restricted and repetitive motor patterns in autism—were significantly curtailed in treated animals.
- Anxiety and Novelty Neophobia: In elevated plus-maze assays, DHA-treated rats made more frequent entries and spent more overall time within the open, exposed arms, signaling reduced anxiety-like inhibition. In fasting latency tests, treated rats exhibited faster approaches to food and increased dietary consumption.
The JNK Molecular Target and Prefrontal Antioxidant Defenses
To map the biochemical signaling networks mediating these behavioral recoveries, the team integrated network pharmacology with molecular docking simulations, Western blotting, and enzyme-linked immunosorbent assays (ELISA).
Network analysis revealed 13 core overlapping genetic targets between DHA bioactivity and ASD pathology, notably involving genes encoding tumor necrosis factor (TNF), interleukin-6 (IL6), superoxide dismutase 1 (SOD1), catalase (CAT), brain-derived neurotrophic factor (BDNF), and peroxisome proliferator-activated receptor alpha (PPARA). Computational docking simulations predicted strong direct binding between DHA and key mediators, including JNK, CAT, SOD1, TNF-α, IL-6, and heme oxygenase-1 (HO-1), with binding affinities ranging from −2.93 to −6.02 kcal/mol.
Biochemical analysis of prefrontal cortex tissue validated these computational predictions:
- Selective JNK Inactivation: DHA administration suppressed the phosphorylation of JNK (reducing p-JNK) and downstream c-JUN transcription factor activation. Crucially, total JNK levels remained unchanged, confirming that the fatty acid silences pathological enzymatic activation rather than disrupting basal protein expression.
- Suppression of Pro-inflammatory Cytokines: Downstream of JNK inhibition, the prefrontal cortex exhibited marked reductions in prominent neuroinflammatory cytokines, including TNF-α, IL-6, and interleukin-1 beta (IL-1β).
- Restoration of Antioxidant Capacities: Simultaneously, DHA augmented nuclear factor erythroid 2-related factor 2 (NRF2) signaling, driving significant upregulation of endogenous antioxidant enzymes including HO-1, SOD1, and CAT to clear neurotoxic reactive oxygen species.
Preclinical Caveats and Clinical Horizons
The authors emphasize that while these findings outline a clear molecular framework connecting omega-3 supplementation to reduced neuroinflammation and improved social behavior, the results remain strictly preclinical.
Because the study utilized a neonatal stress-induced animal model, the results cannot be directly extrapolated to human clinical cohorts, nor do they suggest that omega-3 fatty acids can replace established clinical interventions, speech therapies, or behavioral supports.
Future translational research will require carefully stratified clinical trials with rigorous behavioral metrics and biomarker validation to determine whether specific neurodevelopmental subgroups—particularly individuals with verified elevations in baseline oxidative stress or cytokine dysregulation—can experience clinical gains from targeted adjunctive DHA therapies.
Funding Information
National Natural Science Foundation of China (82071395, 82101466, 32371030); Chongqing Medical University Program for Youth Innovation in Future Medicine (W0044); Doctoral Initiation Fund of Women and Children’s Medical Center of Guangzhou Medical University (1600037-04); Science and Technology Program of Guangzhou (2024A03J1244).
Editorial Notes:
- This article was edited by a Neuroscience News editor.
- Journal paper reviewed in full.
- Additional context added by our staff.
About this autism Research:
- Media Contact: Shuai Ding
- Source: Chinese Academy of Sciences
- Image Credit: Image credited to Neuroscience News
- Original Research is Open Access: Pediatric Discovery (Oct 1, 2026). “Docosahexaenoic acid ameliorates autistic-like behaviors by inhibiting oxidative stress and inflammatory response in neonatal maternal separation rats.” Authors: Boqing Xu, Hao Yuan, Xiaohuan Li, Qingyang Yu, Chunfang Dai, Zhifang Dong.
- DOI: 10.1002/pdi3.91
Abstract
Docosahexaenoic acid ameliorates autistic-like behaviors by inhibiting oxidative stress and inflammatory response in neonatal maternal separation rats
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.
