Autism spectrum disorder (ASD) comprises a group of neurodevelopmental disorders whose etiology and pathogenesis are not fully understood. To gain insight into the molecular basis of ASD, we performed comparative integrated proteomic and metabolomic analyses of urine samples from children diagnosed with ASD and healthy children. All 160 samples underwent proteomics analysis and 60 were analyzed by liquid chromatography-mass spectrometry to obtain metabolite profiles. We identified 77 differentially expressed proteins (DEPs; 21 downregulated and 56 upregulated) and 277 differentially expressed metabolites; 31 of the DEPs including glutathione, leukocyte antigens, glycoproteins, neural adhesion factors, and immunoglobulins, have been implicated in neuroinflammation. The proteomic analysis also revealed 8 signaling pathways that were significantly dysregulated in ASD patients; 3 of these (transendothelial leukocyte migration, antigen processing and presentation, and graft vs. host disease) were associated with the neuroimmune response. The metabolism of tryptophan, which is also related to the neuroimmune response, has been found to play a potential role in ASD. Integrated proteome and metabolome analysis showed that 6 signaling pathways were significantly enriched in ASD patients, 3 of which were correlated with impaired neuroinflammation (glutathione metabolism, metabolism of xenobiotics by cytochrome P450 and transendothelial migration of leukocyte). We also found a correlation between prostaglandin (PG) E2 levels and the inflammatory response in ASD. These results underscore the prominent role of the neuroimmune response in ASD and provide potential biomarkers that can be used for diagnosis or as targets for early intervention.

The exact cause of Autism spectrum disorder (ASD) remains unclear. The accumulation of heavy metals and the imbalance of trace elements are believed to play a key role in the pathogenesis of ASD. This study aimed to compare the levels of trace elements and heavy metals in the hair of 1–16-year-old children with varying ASD severity. We included a control group of 57 children, as well as 124 children with autism, consisting of 53 with mild to moderate autism and 71 with severe autism. Questionnaires and hair samples were collected, and 21 chemical elements were analyzed using inductively coupled plasma mass spectrometry (ICP-MS). Children with severe autism showed significantly higher levels of the trace elements copper (Cu) and heavy metals vanadium (V), cobalt (Co), nickel (Ni), arsenic (As), cadmium (Cd), and lead (Pb) in their bodies compared to the control group. Boys with severe autism showed significantly higher levels of Cu, As, Cd, and Pb compared to the control group, while girls with severe autism exhibited significantly lower levels of potassium (K) and increased levels of Pb. Severely autistic children under 7 years old had significantly increased levels of Mn, Cu, V, Co, Ni, As, Cd, and Pb. Children with severe autism aged 7–16 years typically showed significantly higher levels of Cu and As. These findings underscore the importance of heavy metals and essential minerals as environmental factors in the severity of ASD disease.

Autism spectrum disorder (ASD) is a multifactorial neurodevelopmental disorder, with lead (Pb) exposure increasingly linked to its risk. However, the molecular mechanisms linking Pb to ASD remain poorly understood. This study established a postnatal Pb-exposed mouse model and employed the three-chamber social test and the marble-burying test to assess ASD-like behavioral phenotypes. The Pb levels in both blood and the hippocampus were quantified, and hippocampal neurons were assessed for morphological alterations. Moreover, a Tandem Mass Tag (TMT)-based quantitative proteomics approach was applied to elucidate the underlying mechanisms. Neurobehavioral experiments revealed Pb-exposed C57BL/6 offspring exhibited reduced social interaction and novelty preference along with increased repetitive marble-burying behavior. The Pb levels in both the blood and hippocampus of Pb-treated mice were significantly elevated compared with those of control animals. Postnatal Pb exposure resulted in a reduction in the neuronal numbers and disorganized neuronal arrangement in the hippocampus. A total of 66 proteins were identified as being differentially expressed after postnatal Pb exposure. Among them, 34 differentially expressed proteins were common in both Pb exposure groups, with 33 downregulated and 1 upregulated. Bioinformatic analysis revealed multi-pathway regulation involved in Pb-induced neurodevelopmental disorders, including dysregulation of synaptic signaling, abnormal activation of neuron apoptosis, and neuroinflammation. Notably, the SYT10/IGF-1 signaling pathway may play a potential key role. These findings enhance understanding of Pb-induced autism-like behaviors, providing novel proteomic insights into the etiology of ASD.

Autism spectrum disorder (ASD) is a complex neurodevelopmental condition characterized by impairments in social communication and repetitive behaviors, involving various brain regions. Emerging evidence highlights the critical role of the cerebellum in the pathophysiology of autism; however, the underlying molecular mechanisms remain poorly understood. This study aimed to establish a prenatal valproic acid (VPA)-induced mouse model of ASD and explore the potential molecular mechanisms underlying cerebellar ASD-like phenotypes through DIA-based proteomics and bioinformatics analyses. Significant cognitive impairment and anxiety-like behaviors were detected using an open field test and novel object test following VPA exposure, respectively. Additionally, reduced numbers of Purkinje cells with irregular arrangement were observed in the cerebellum. Furthermore, cerebellar proteomics analyses revealed that they identified 193 differentially expressed proteins (DEPs) involved in multiple pathways, including axon guidance, glutamatergic synapse, long-term potentiation, and calcium signaling, among others. Notably, dysfunction of glutamate receptor signaling and disruptions in axon-guidance signaling appear to be major molecular mechanisms underlying cerebellar impairment. Together, these findings suggest that Grin2b may serve as a critical molecule linking synaptic neurotransmission and neurodevelopmental disorders. Thus, Grin2b may represent a potential therapeutic target for addressing cognitive impairment in ASD.

Autism spectrum disorder (ASD) is a neurodevelopmental condition marked by social/communication deficits and behavioral abnormalities, with neuronal apoptosis and immune-inflammatory dysregulation implicated in its pathogenesis. Marine-derived polysaccharides, particularly those from Enteromorpha prolifera (PEPs), exhibit neuroprotective and anti-inflammatory properties—yet their therapeutic potential for ASD remains unexplored. Major monosaccharide components of PEPs were identified as rhamnose, xylose, glucose, glucuronic acid, galactose, and ribose through ion chromatography analysis. Infrared spectroscopy confirmed PEPs as pyranose-type polysaccharides with α-glycosidic bonds and uronic acids, while gel permeation chromatography showed a predominant molecular weight of 3.813 kDa (83.919%). To explore the therapeutic potential of PEPs in ASD, a comprehensive method combining network pharmacology, molecular docking, and in vitro validation was conducted. A total of 235 ASD-related target proteins were predicted, with enrichment analyses indicating significant involvement in pathways such as neuroactive ligand–receptor interaction and the MAPK signaling pathway. In vitro assays using valproic acid (VPA)-induced HT22 neuronal cells showed that PEPs significantly attenuated apoptosis. Western blot analysis further confirmed the downregulation of HSP90AA1, cleaved CASP3/pro-CASP3, p-NF-κB1/NF-κB1, p-AKT1/AKT, and p-mTOR/mTOR, as well as the upregulation of IκBα after PEPs treatment. These findings suggest that PEPs exert neuroprotective effects through the modulation of apoptosis and inflammation-related signaling pathways, supporting their potential as a promising candidate for further study in ASD.

Background: Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder, with prenatal exposure to valproic acid (VPA) being a recognized environmental risk factor for ASD, closely associated with its neurotoxic mechanism and oxidative stress. Marine green algae, such as Ulva prolifera , are rich sources of bioactive polysaccharides known for their antioxidant, anti-inflammatory, lipid-lowering, anti-tumor, and neuroprotective properties, with a core focus on antioxidant effects. Therefore, this study aims to investigate the protective effects and potential mechanisms of Ulva prolifera polysaccharides (PUPs) on oxidative stress damage induced by VPA in mouse hippocampal neuronal HT22 cells. Method: The primary structure of PUPs was determined through infrared spectroscopy, liquid chromatography, and gel permeation chromatography. PUPs intervention in the VPA-induced HT22 oxidative damage cell model was used to measure oxidative stress factor levels. Results: PUPs were characterized as composite polysaccharides with α-glycosidic bonds, sulfate ions, and aldonic acids, containing mannose, rhamnose, glucuronic acid, glucose, galactose, and xylose. The main molecular weight (Mw) was 2.124 kDa (80.535%). In vitro experiments revealed that PUPs significantly increased cell viability in the VPA model, enhanced intracellular SOD and CAT activities to boost antioxidant capacity, and concurrently reduced ROS levels and MDA content. Western blot analysis revealed that PUPs increased Nrf2, HO-1, and IκBα levels, while decreasing the expression levels of Keap1, p-NF-κB-p65/NF-κB-65, p-Erk/Erk, p-p38/p38, and p-JNK/JNK. Conclusions: PUPs demonstrate a significant protective effect against VPA-induced oxidative stress damage in HT22 neuronal cells. As a natural antioxidant, PUPs hold promising potential for the prevention and adjunct treatment of ASD-related neuronal oxidative stress damage.