Glutaric acidemia type 1 (GA I) is caused by deficient activity of glutaryl-CoA dehydrogenase, leading to predominant accumulation of glutaric acid (GA) in the brain. GA I patients present progressive neurological deterioration whose pathophysiology is only partially elucidated. We investigated whether intracerebral GA administration, alone or combined with quinolinic acid (QA), a pro-inflammatory intermediate of the kynurenine pathway, could alter the expression of genes associated with inflammatory signaling, endoplasmic reticulum (ER) stress, and neurotrophic support in cerebral cortex of wild-type (WT) and Gcdh−/− mice. We also tested the effects of L-carnitine (Carn) on these parameters. GA alone increased mRNA levels of the genes encoding NF-κB (NFKB1), COX-2 (PTGS2), iNOS (NOS2), and TLR2 (TLR2) in both genotypes, while reducing those of IL-10 (IL10) and VEGF-A (VEGFA) only in Gcdh−/− mice. Combined QA + GA treatment produced a larger response, including increased TNF-α (TNF), IL-1β (IL1B), IL-6 (IL6), NLRP3 (NLRP3), CHOP (DDIT3) and PERK (EIF2AK3) mRNA levels in the Gcdh−/− mice, besides reducing IκBα (NFKBIA), IL-10 (IL10) and BDNF (BDNF) expression in both genotypes, and VEGF-A (VEGFA) in the Gcdh−/− mice. We also found that lysine (Lys) and QA treatment elevated TNF-α, IL-1β, and IL-6 protein levels in Gcdh−/− mice. Carn prevented or attenuated most transcriptional alterations induced by QA + GA and reduced cytokine protein elevations elicited by Lys + QA administration. These findings indicate that Carn modulates acute molecular responses related to inflammation, ER stress, and neurotrophic factors in the cerebral cortex of the GAI mouse model.
Maple Syrup Urine Disease (MSUD) is caused by a genetic mutation in the branched-chain α-ketoacid dehydrogenase complex, resulting to accumulation of branched-chain amino acids (BCAAs) that affect the central nervous system and cause neurochemical alterations and behavioral changes. In this line, full-spectrum cannabidiol (CBD)-rich oil has emerged as a potential therapeutic strategy. Therefore, this study aims to evaluate the effects of two doses of the compound full-spectrum CBD-rich oil in a BCAA-induced MSUD rat model, against behavioral, cholinergic, inflammatory, and oxidative stress parameters. For this, animals were divided into six groups: control group, CBD 3.5 mg/kg group, CBD 7.5 mg/kg group, BCAA group, BCAA + CBD 3.5 mg/kg group, and BCAA + CBD 7.5 mg/kg group. The treatment was administered over 21 days; after that, the animals were subjected to open-field and object recognition tests. Next, we extracted the cerebral cortex to analyze cholinergic function, inflammation, and oxidative stress. The results show that the open-field test revealed no differences in crossings and rearings across all groups. In object recognition test, control and CBD 3.5 groups showed improved short- and long-term memory compared to training. The CBD 7.5 and BCAA + CBD 3.5 groups showed improvement only in short-term memory. BCAA control and BCAA + 7.5 did not present differences. In the cholinergic system, BCAA control showed decreased choline acetyltransferase (ChAT) activity, which was reversed by CBD treatment at both doses. The BCAA + CBD 7.5 shows increased ChAT activity compared to control group. While acetylcholinesterase (AChE) was reduced in the CBD 7.5 groups and increased in the BCAA control group, both CBDs reversed this increase in BCAA control group. Inflammatory cytokines show increased interleukin-1β in BCAA control group, and the CBD treatment decreases its levels compared to BCAA and saline control groups. Interleukin-6 increases in BCAA control group, and CBD 3.5 reverses it. Tumoral necrosis factor-alpha was reduced in BCAA + CBD 3.5 and BCAA + CBD 7.5 compared to control and BCAA control groups. Further, under oxidative stress, BCAA control increases 2,7-dichlorofluorescein oxidation and thiobarbituric acid levels, which were reversed by CBD treatment. Sulfhydryl content was decreased in CBD 7.5, BCAA control group, BCAA + CBD 3.5, and BCAA + CBD 7.5 compared to control group. Superoxide dismutase activity increased across all groups, whereas catalase activity decreased in the BCAA control group; treatment with CBD 7.5 reversed this reduction. Overall, we conclude that full-spectrum CBD-rich oil shows therapeutic potential for MSUD, although optimal dosing and treatment duration require further investigation.
Vascular cognitive impairment (VCI) is a vast disease with chronic cerebral hypoperfusion (CCH) being the major driver. In our previous study, we have verified that Da ChuanXiong (DCX) formula could alleviate VCI, and obviously upregulate the decreased cerebral blood flow, indicating its potential benefits on vascular remodeling. This study aims to explore the pro-angiogenic and vasodilatory effects of DCX formula after CCH injuries, and clarify its pharmacodynamic substances and potential mechanisms. The rat model of CCH was established by bilateral common carotid artery occlusion. CD31+ immunofluorescent staining and quail chorioallantoic membrane (qCAM) experiments were conducted to assess the angiogenic effect of DCX. Isolated vascular ring experiments and NO assay in qCAM were performed to evaluate the vasodilatory effect of DCX. Then we analyzed the chemical components absorbed in the blood of DCX under CCH conditions by UPLC-Q-TOF/MS, and further combined network pharmacology, molecular docking to predict the potential mechanisms and effective components. Western blot was performed in CCH rats to verify the possible mechanisms and in vitro pharmacodynamic verification experiment was conducted for innovative active ingredients in DCX. Our results revealed that DCX obviously increased the number of CD31+ cells in the hippocampus and brain cortex of CCH rats, and significantly increased the vascular areas in qCAM. DCX significantly dilated the pre-constricted vascular ring and increased NO contents in qCAM. Besides, Senkyunolide A, Butylphthalide, Ferulic acid, Senkyunolide I, Levistilide A, Ligustilide, Senkyunolide H and Gastrodin were identified as the 8 major blood-absorbed compounds of DCX under CCH condition. Next, network pharmacology and molecular docking revealed NOS3 and PI3K/Akt signaling as the critical targets and pathway of DCX improving vascular remodeling against CCH injuries, which was further validated by western blot. Moreover, we firstly discovered that Senkyunolide A and Senkyunolide H could obviously promote cell migration and enhance the tube formation ability of HUVECs in vitro. Taken together, DCX formula effectively enhanced the angiogenesis and vasodilation via activating PI3K/Akt/eNOS signaling, which might be the important pathways for DCX promoting vascular remodeling against CCH injuries. Notably, Senkyunolide A and Senkyunolide H were firstly identified as innovative components responsible for the pro-angiogenic effects of DCX.
Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder with increasing global prevalence but a lack of reliable diagnostic biomarkers. Emerging evidence suggests that immune dysregulation, gut–brain axis dysfunction, and increased intestinal permeability play key roles in ASD pathophysiology. This study investigated the combined diagnostic value of ADAM10 and cytokines (IL-10, IL-22, IL-17 A, and IL-17D). Multivariable logistic regression produces an improved ROC curve that improves diagnostic accuracy over individual markers by combining numerous predictors into a single risk score (linear predictor). The technique, which frequently raises individual marker AUCs, entails modelling a binary result, calculating the probability, and visualizing ROC based on the projected probabilities. In this case–control study, plasma levels of ADAM10, IL-10, IL-22, IL-17 A, and IL-17D were measured in 37 male children with ASD and 37 age-matched controls. Group comparisons, correlation analyses, and receiver operating characteristic (ROC) curve analyses, including combined ROC models, were performed. ADAM10, IL-22, and IL-17 A levels were significantly reduced in children with ASD compared to controls, whereas IL-10 and IL-17D showed no significant differences. ADAM10, IL-17 A, and IL-22 demonstrated good diagnostic performance, with AUC values of 0.886, 0.855, and 0.812, respectively. In contrast, IL-10 and IL-17D showed poor discriminatory ability, with AUC values of 0.524 and 0.599, respectively. Combined ROC analysis markedly improved diagnostic accuracy, with all panels including ADAM10 achieving AUC values above 0.90, and some reaching as high as 0.988, with high sensitivity and specificity. The combination of ADAM10 with selected cytokines significantly enhances diagnostic performance compared to individual markers, supporting a link between immune dysregulation, barrier dysfunction, and gut permeability in ASD.
This study aimed to investigate whether low-intensity pulsed ultrasound (LIPUS) targeted at the liver can alleviate cerebral Aβ pathology by improving peripheral metabolism and enhancing hepatic Aβ clearance in a comorbid mouse model. 5xFAD transgenic mice were fed a high-fat diet (HFD) to induce peripheral IR. Mice received hepatic LIPUS intervention for four weeks. Outcomes were assessed using metabolic tests, in vivo and ex vivo fluorescence imaging of Aβ trafficking, neuropathological analysis, proteomics, and the Morris water maze. LIPUS ameliorated HFD-induced IR, hepatic steatosis, and pancreatic islet hyperplasia. It upregulated hepatic expression of Aβ-clearance proteins (IDE, LRP-1), enhanced peripheral Aβ catabolism, and reduced Aβ infiltration into the brain. Consequently, LIPUS attenuated cerebral Aβ deposition, neuronal apoptosis, and partially rescued spatial learning and memory deficits. Proteomic analysis confirmed systemic upregulation of metabolic and clearance pathways. Hepatic LIPUS modulates the peripheral metabolism–central clearance axis, reducing brain Aβ burden and improving cognitive function. This non-invasive approach presents a novel strategy for AD, particularly in the context of metabolic dysfunction.
Alzheimer’s disease is increasingly recognized as a multifactorial neurodegenerative disorder characterized by complex interactions among metabolic dysfunction, chronic inflammation, mitochondrial impairment, and lipid dysregulation. Although the amyloid cascade hypothesis has long dominated Alzheimer’s disease research, limited clinical success of amyloid-targeted therapies has highlighted the need for broader mechanistic frameworks integrating systemic metabolic and cellular dysfunction. Emerging evidence suggests that obesity-associated phospholipid remodeling, particularly dysregulation of phosphatidylethanolamine (PE), may critically influence neurodegenerative progression through alterations in membrane dynamics, mitochondrial homeostasis, autophagy, oxidative stress, and neuroimmune signaling. This review critically examines the molecular and cellular mechanisms underlying PE-mediated adipose–brain crosstalk in obesity-associated Alzheimer’s disease, with emphasis on mitochondrial dysfunction, blood–brain barrier disruption, ferroptosis, membrane remodeling, and microglial activation. The review further discusses how altered PE metabolism may impair synaptic integrity, promote lipotoxicity, and enhance neuronal vulnerability under chronic metabolic stress. Current evidence, however, remains constrained by methodological heterogeneity, inconsistent lipidomic findings, and substantial reliance on experimental animal models, limiting translational interpretation. In addition, this review proposes the concept of a “metabolic membrane remodeling axis” as an integrative framework linking obesity-driven phospholipid dysregulation with neuroimmune and neurodegenerative progression. A deeper understanding of PE-driven neuroimmunometabolic dysfunction may facilitate biomarker discovery and support the development of lipid-targeted therapeutic strategies for obesity-associated Alzheimer’s disease. Proposed “metabolic membrane remodeling axis” illustrating how obesity-driven phosphatidylethanolamine (PE) dysregulation may promote neuroimmune remodeling, mitochondrial dysfunction, oxidative stress, synaptic degeneration, and progressive neurodegeneration in Alzheimer’s disease while highlighting current translational limitations and future lipidomics-based therapeutic opportunities.
Hepatic encephalopathy (HE) is a complex neuropsychiatric syndrome characterized by a spectrum of symptoms ranging from mild cognitive and motor disturbances to severe coma. γ-Oryzanol (ORY), a natural compound derived from rice bran, has demonstrated potential to enhance memory and cognitive function in preclinical studies. This study investigated the neuroprotective and cognitive-enhancing effects of γ-oryzanol in a rat model of HE induced by bile duct ligation (BDL). Male Wistar rats underwent BDL surgery and received either γ-oryzanol (100 mg/kg) or vehicle via oral gavage for subchronic 28 days. Four experimental groups were included: sham-operated, sham + ORY, BDL, and BDL + ORY. Cognitive function was assessed using the Novel Object Recognition (NOR) test and Y-maze. Blood samples were analyzed for oxidative stress markers (antioxidant status, lipid peroxidation, thiol levels), liver function markers (bilirubin, albumin, AST, ALT, ALP, LDH), and prefrontal cortex tissues were examined histologically. BDL induction significantly impaired cognitive performance and increased oxidative stress and neuronal damage in the prefrontal cortex. γ-Oryzanol treatment effectively restored cognitive function, reduced oxidative stress, and attenuated neuronal damage without significantly altering liver enzyme levels. These findings suggest that γ-oryzanol exerts neuroprotective effects in HE possibly through antioxidant-related pathways, although this mechanism was not consistently supported by all oxidative stress markers. The protective effects appeared largely independent of major liver function improvement.
Parkinson’s disease (PD) is the world’s fastest-growing neurodegenerative disorder, presenting a significant burden on patients, caretakers, and healthcare systems. Despite advances in symptomatic management, disease-modifying therapies remain inaccessible, underscoring the need to focus on prevention and neuroprotection. Estrogen has emerged as a potential neuroprotective agent due to its diverse effects on neuronal survival and function. This review examines the neuroprotective role of estrogen in the pathogenesis of PD. A comprehensive literature search was conducted using the PubMed, Scopus, and Web of Science databases, with a focus on studies published within the last 10 years. The combinatorial keyword strategies, including “estrogen,” “parkinson’s disease,” “estrogen receptor alpha (ERα),” “estrogen receptor beta (ERβ)”, “G-protein coupled estrogen receptor 1 (GPER1),” “neurodegeneration,” “oxidative stress,” “neuroinflammation,” “neuroprotection,” and “preclinical studies.” Findings indicate that estrogen deficiency promotes Parkinson’s-like pathology, which includes oxidative stress, mitochondrial impairment, synaptic dysfunction, and neuroinflammation. Activating ERα, ERβ, and GPER1 through both genomic and non-genomic mechanisms slows disease progression by reducing oxidative damage, inflammatory cytokine release, and neurodegeneration. Preclinical evidence suggests that several natural, synthetic, and semisynthetic estrogen compounds protect neurons by modulating PI3K/Akt, MAPK/ERK, and NF-κB signalling pathways, thereby reducing dopaminergic neuronal loss and improving behavioural outcomes. Estrogen protects the brain from neurotoxic insults, while a deficiency of estrogen exacerbates oxidative stress, mitochondrial dysfunction, and neuroinflammation. While activation of ERα, ERβ, and GPER promotes antioxidant, anti-inflammatory, and anti-apoptotic effects, reducing PD pathogenesis. These findings support estrogen receptors as potential therapeutic targets for PD management.
Huntington’s disease (HD) is a neurological condition with limited treatment. Its hallmarks are progressive loss of neurons, chorea, cognitive, motor, and metabolic impairments. The current study uses 3-nitropropionic acid (3-NP) to cause HD-like indications in rats. L-theanine (L-TH), an active component of Camellia sinensis, has already been shown to possess anti-inflammatory and anti-oxidant effects. The impact of the isolated phytoconstituent, L-TH, on 3-NP-induced changes in three distinct brain regions was assessed in the current study. The animals received a 7-day pretreatment consisting of normal saline, extract of C. sinensis (100 and 200 mg/kg b.w., i.p.), L-TH (10 mg/kg b.w., i.p.), and standard Tetrabenazine (TBZ, 25 mg/kg, b.w., i.p.). The administration of 3-NP (10 mg/kg b.w., i.p.) to the treatment groups began on the 8th day of the protocol and continued until the 21st day. The brain homogenates underwent biochemical and neurochemical assessments. In conclusion, L-TH improved behavioral outcomes and attenuated 3-NP-induced neurochemical alterations in brain homogenates. Histopathological analysis revealed lower levels of tissue damage in the L-TH-treated group compared to the 3-NP group. Molecular docking studies further indicated favorable interactions of L-TH with the HTT protein. These findings suggest that L-TH may attenuate 3-NP-induced Huntington’s disease-like alterations in rats. However, additional studies using genetic HD models and molecular mechanisms are required to confirm its therapeutic efficacy and underlying mode of action. A schematic representation of the isolation of L-theanine (L-TH) from the hydroalcoholic extract ofCamellia sinensis (HECS) and the evaluation of its protective effect against 3-NP-induced HD-like alterations in a 3-nitropropionic acid (3-NP) model
Background and aims: The relationship between type A hepatic encephalopathy, a highly lethal disease, and gut microbiota remains unclear, and research on this topic is limited. The objective of our study was to investigate the correlation between an imbalance in the gut microbiota and type A hepatic encephalopathy and the impact of fecal microbiota transplantation. Methods: We established a mouse model of gut microbiota disorder and type A hepatic encephalopathy. Feces from grades III and IV type A hepatic encephalopathy mice were transplanted into healthy mice. Antibiotic administration, intestinal symbiosis, and pathogenicity experiments were conducted. Behavioral, biochemical, pathological, 16 S rRNA gene amplicon sequencing analyses, and correlation analyses were performed. Results: Antibiotic treatment caused a gut microbiota imbalance in mice. The degree of thioacetamide-induced type A hepatic encephalopathy was significantly aggravated after oral antibiotic administration, leading to a decline in the survival curve, accompanied by behavioral, biochemical, and pathological changes, as well as decreased Rikenellaceae levels. Transplanting feces from type A hepatic encephalopathy mice into healthy mice resulted in thioacetamide-like behavioral, biochemical, and tissue changes, as well as a significant decline in the abundance of the gut microbiota, an increase in the abundance of Prevotellaceae NK3B31, and a decrease in the abundance of Akkermansia muciniphila and Odoribacter. Additionally, significant correlations were observed between the abundances of the four intestinal microbial species and the majority of measured indicators in mice with type A hepatic encephalopathy. Notably, Akkermansia muciniphila exhibited particularly strong associations with these indicators. Although significant between-group differences were observed for Bacillus, Paenibacillus, Candidatus Saccharimonas, Escherichia-Shigella, UCG_002, Acinetobacter, and Proteus, no significant correlations were detected between these microbial taxa and any of the measured indicators. Conclusions: Gut microbiota disorder aggravates lesions in thioacetamide-induced type A hepatic encephalopathy mice. Transplanting feces from mice with type A hepatic encephalopathy causes healthy mice to exhibit type A hepatic encephalopathy symptoms.
Type 2 diabetes mellitus impairs brain metabolic and mitochondrial homeostasis, yet the intensity-dependent neuroprotective effects of exercise remain poorly defined. This study examined the intensity‑dependent effects of interval training on brain mitophagy, metabolic signaling, oxidative stress, and neuroinflammation in a type 2 diabetes model. Fifty male rats were assigned to five groups (n = 10 each): healthy control (HC), diabetic control (DC), diabetic + low-intensity interval training (LIIT), diabetic + moderate-intensity interval training (MIIT), and diabetic + high-intensity interval training (HIIT). Following the intervention, hippocampal and cortical tissues were analyzed for metabolic signaling markers (AMPK, ULK1, mTOR), mitophagy-related proteins (PINK1, Parkin, LC3-II/I, p62), oxidative and antioxidant indices (MDA, SOD, CAT, TAC), pro-inflammatory cytokines (TNF-α, IL-6, IL-1β), and lipid peroxidation (4-HNE). Statistical significance was set at p < 0.05. MIIT and HIIT activated AMPK–ULK1 signaling and suppressed mTOR in the hippocampus and cortex, leading to enhanced mitophagy, particularly in the hippocampus. These adaptations were accompanied by improved redox balance, reduced lipid peroxidation, and attenuated neuroinflammation, with effects increasing in an intensity-dependent manner (LIIT < MIIT < HIIT). Interval training may support neuroprotective adaptations in type 2 diabetes, with moderate- and high-intensity protocols potentially offering greater benefits than low intensity. Still, these effects require confirmation in broader experimental contexts.
Therapeutic hypothermia (TH) is the standard treatment to reduce mortality and neurological impairments after neonatal hypoxia–ischemia (HI). However, TH does not fully suppress neuroinflammation, leading to persistent neuroinflammatory responses that may exacerbate brain injury and limit TH efficacy. In addition, the influence of sexual dimorphism on these outcomes remains poorly understood. We investigated short- and long-term microglial and astrocytic reactivity in male and female neonatal rats following HI and TH treatment. At postnatal day 7 (PND7), male and female pups underwent right common carotid occlusion followed by exposure to hypoxia (8
Neurological disorders, including stroke, Parkinson’s disease (PD), and Alzheimer’s disease (AD), are among the most prevalent and debilitating neuropathological conditions, affecting over 50 million people globally. Effective prevention and treatment strategies require a comprehensive understanding of reliable and disease-specific biomarkers. Such insights can enhance diagnostic precision, enable early detection, and support the development of targeted therapeutic approaches. In this review, we focus on exosome-derived proteins and small non-coding RNAs as promising non-invasive biomarkers for stroke, PD, and AD, given the accessibility and stability of exosomes in biological fluids. Furthermore, we examine their potential diagnostic and therapeutic relevance, emphasizing their functional roles, specificity, and sensitivity. Finally, we discuss current limitations, emerging challenges, and future research directions to guide the advancement of exosome-based biomarker discovery and clinical translation in neurological disorders. However, we emphasize that the comparative effectiveness of exosomal microRNAs (miRNAs) versus protein biomarkers requires direct validation in future head‑to‑head cohort studies. Conceptual framework linking the pathophysiological basis of neurological disorders to exosome-based diagnostics and therapeutics. In Alzheimer’s disease (AD), Parkinson’s disease (PD), and stroke, neurodegeneration is driven by pathological molecules such as amyloid-β (Aβ), α-synuclein, tau, NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome components, and disease-associated microRNAs. These factors are actively secreted within exosomes from affected neurons into the bloodstream. The minimally invasive isolation of circulating exosomes enables biomarker assays that can facilitate early diagnosis, a critical window when neuroprotective interventions are most effective. Furthermore, based on the exosomes’ content, targeted therapeutics, including small non-coding RNA replacement or antagonists, antibodies, and gene therapy, can be designed. This dual diagnostic–therapeutic potential highlights exosome biology as a promising frontier for improving clinical outcomes in neurodegenerative and cerebrovascular diseases.
Hepatic encephalopathy (HE) represents a severe neuropsychiatric manifestation secondary to advanced hepatic dysfunction, predominantly driven by hyperammonaemia and associated neuroinflammatory processes. Although lactulose remains a mainstay in the clinical management of HE due to its ammonia-lowering properties, its limited efficacy in attenuating neuroinflammation necessitates the use of adjunctive therapeutic strategies. Previously, we identified berberine, a plant-derived isoquinoline alkaloid, as an anti-HE agent via the reduction of neuroinflammation and NLRP3 inflammasome activation. Therefore, in the present study, we investigated the potential beneficial effects of co-administering berberine and lactulose in a rat model of thioacetamide (TAA)-induced hepatic encephalopathy. HE was induced via intraperitoneal injections of TAA (200 mg/kg) on alternate days for one week. Animals received oral pre-treatment with lactulose (8 mL/kg) alone or in combination with berberine (100 mg/kg) for 14 consecutive days. Comprehensive behavioural assessments were conducted alongside biochemical quantification of systemic ammonia, hepatic transaminases (ALT, AST), and oxidative stress markers (MDA, GSH) in both hepatic and cerebral tissues. Histopathological evaluations of the liver and brain further corroborated the biochemical findings. Notably, combination therapy significantly ameliorated cognitive deficits, reduced systemic ammonia burden, normalised liver and brain enzyme profiles, and markedly attenuated central neuroinflammatory signalling, as evidenced by reduced hippocampal NF-κB activation and GFAP immunoreactivity, in addition to suppressing hepatic NF-κB expression, surpassing the effects observed with monotherapy. Histological analyses revealed reduced neuronal degeneration and hepatic architectural disruption. These findings underscore the therapeutic potential of berberine-lactulose co-treatment in mitigating both hepatic and neuroinflammatory pathologies associated with HE, offering a promising adjunctive strategy for improving neurocognitive outcomes in liver failure. Berberine potentiates lactulose efficacy in thioacetamide-induced hepatic encephalopathy. Combination therapy reduces hyperammonemia and improves neurobehavioral deficits. Berberine-lactulose co-treatment attenuates hepatic injury, fibrosis, and oxidative stress. Combination therapy suppresses hippocampal NF-κB activation and reactive astrogliosis while reducing hepatic inflammation. Combined therapy offers superior hepato-neuroprotection over lactulose monotherapy.
Peripheral nerve injury-induced persistent pain hypersensitivity cannot be fully explained by increased neuronal excitability alone. Rather, it is increasingly recognized as a pathological tissue state characterized by sustained glial activation, immune signaling, and synaptic remodeling within the spinal cord. In this context, metabolic remodeling of the spinal cord has emerged as a potentially important feature of neuropathic pain. However, the metabolic patterns associated with this state, and their relationship to underlying molecular programs, remain incompletely defined. Here, we performed GC–MS-based untargeted metabolomic profiling of the spinal cord dorsal horn on day 7 after spinal nerve transection (SNT). To provide orthogonal validation, we integrated pathway-analysis results from four independent spinal cord RNA-sequencing datasets derived from distinct neuropathic pain models and further conducted qPCR-based validation. Metabolic profiling revealed a clear separation between SNT and sham samples, marked by broad depletion of the free fatty acid pool and features consistent with an immunometabolic shift. Consistently, analyses across RNA-sequencing datasets and qPCR validation demonstrated upregulation of immune and inflammatory programs, together with downregulation of fatty acid metabolism and cholesterol homeostasis. Collectively, these findings suggest that persistent neuropathic pain should be interpreted not simply as a consequence of neuronal and immune signaling, but also through the metabolic tissue environment that supports and sustains this pathological state.
Majoon-e-Boolis (MB), a traditional Unani formulation, has been used for neurological conditions; however, its molecular mechanism remains largely unexplored. The current study was designed to explore the novel mechanistic insights between antioxidant activity, anti-inflammatory effects, dopaminergic support and anti-aggregation properties of the formulation for modulating the pathogenesis of Alzheimer’s Disease (AD). Behavioral deficits were studied via Novel Object Recognition, Elevated Plus Maze and Open Field Test. In addition, the study looked at the immunohistochemistry of superoxide dismutase 2, tumor necrosis factor-alpha, tyrosine hydroxylase and α-synuclein (α-Syn) against Aβ1–42-induced AD model. Additionally, histopathological alterations were also performed against the AD model. The observed findings demonstrated that MB treatment in Aβ1–42-induced AD in rat models significantly improved their cognitive impairments and alleviated anti-anxiety behaviour with enhanced locomotor activity. Moreover, MB induced significant reduction in oxidative stress and neuroinflammation with enhanced dopaminergic neuronal activity. GC-MS analysis of MB revealed the presence of bioactive phytochemicals such as thiophene, guanidinopropionic acid, 5-hydroxymethylfurfural etc. The findings underscore the potential of MB as a promising therapeutic agent in mitigating the multifaceted pathology of Aβ1–42-induced AD in a rat model by exerting its neuroprotective effects, cognition restoration and overall maintenance of neuronal architecture through a combination of antioxidant, anti-inflammatory and anti-aggregation mechanisms, making it a multifaceted therapeutic candidate for AD.
Ischemic stroke (IS) is a detrimental neurological disorder with limited clinical therapeutic options. The Astragalus-Safflower herb pair (AS) shows therapeutic effects against IS, but its mechanism remains unclear. Herein, this study combined cerebral untargeted metabolomics, mitochondrial ultrastructural observation and 16 S rDNA gut microbiota profiling to systematically explore its neuroprotective mechanism. This study aimed to evaluate the efficacy of AS in alleviating IS, and investigate its mechanism of action. Rat models of middle cerebral artery occlusion (MCAO) were established using the intraluminal filament method. Behavioral tests confirmed that AS significantly improved neurological deficits and motor function. Untargeted metabolomics revealed that AS reversed nine IS-related energy metabolites in brain tissue. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment indicated involvement of nicotinamide metabolism and the TCA cycle. Molecular studies demonstrated that AS regulated mitochondrial energy metabolism via the AMPK/Sirt1/PGC-1α axis. Additionally, 16 S rDNA sequencing showed that AS restructured gut microbiota composition. Correlation analysis suggested a gut-brain axis mechanism linking microbial changes to cerebral energy metabolism. In conclusion, AS exerts neuroprotection against IS by activating the AMPK/Sirt1/PGC-1α pathway, thereby enhancing TCA cycle activity and maintaining energy homeostasis. Concurrently, AS reshapes the gut microbiota, contributing to its effects via the gut-brain axis. Taken together, this integrated multi-omics study systematically clarifies the multi-target pharmacological characteristics of AS. These findings provide a novel multi-target mechanistic basis for the clinical application of AS in the treatment of IS and enrich the understanding of its synergistic neuroprotective mechanism.
Dysregulated lipid metabolism and neuroinflammation are increasingly recognized as interacting contributors to Alzheimer’s disease (AD), but the cell-type-specific genetic links between fatty acid metabolism and AD remain incompletely defined. This study aimed to identify fatty acid metabolism-related genes associated with AD risk and to characterize their relevance to perivascular macrophage (PVM) states. Single-cell RNA-sequencing data from GSE160936 and bulk transcriptomic data from GSE270454 were integrated to evaluate cell-type-specific fatty acid metabolism activity in AD and control samples.The primary data sources, fatty acid metabolism gene set, AUCell scoring strategy, CellChat workflow, GSEA resources, and MR software settings were specified to improve reproducibility. PVMs were further analyzed for differential expression, ligand-receptor communication, pathway enrichment, transcription factor regulation, and pseudotime-associated transcriptional changes. Cis-eQTL-based two-sample Mendelian randomization was performed using eQTLGen exposure data and AD GWAS summary statistics, followed by sensitivity analyses, reverse MR, and Bayesian colocalization.The analyses were interpreted across three distinct evidence levels: cell-type-resolved transcriptional association, systemic genetic expression prioritization, and tissue-level protein expression. Nominal MR findings were interpreted alongside multiple-testing considerations and colocalization support.Candidate protein expression was examined in hippocampal tissue from APP/PS1 and wild-type mice by Western blotting. Single-cell analysis identified eight major cell populations and showed increased fatty acid metabolism activity in PVMs from AD samples. Mendelian randomization prioritized ten fatty acid metabolism-related genes associated with AD risk, among which ACSL1, EPM2AIP1, MALT1, and RASGRP3 showed strong colocalization support (PP.H4 > 0.9). Pathway analyses linked these genes to lipid metabolic regulation, inflammatory signaling, phagocytosis, and mitochondrial/peroxisomal fatty acid metabolism. Co-expression analysis suggested associations between MALT1 and fatty acid oxidation-related genes, including ACADM and ACOX1. Western blotting in whole hippocampal lysates from APP/PS1 mice provided exploratory tissue-level protein evidence, showing increased ACSL1 and MALT1 and decreased RASGRP3 expression; these findings should not be interpreted as confirmatory PVM-specific validation. This integrative analysis prioritizes ACSL1, EPM2AIP1, MALT1, and RASGRP3 as candidate genes connecting fatty acid metabolic dysregulation with AD-associated neuroinflammatory processes. Because the genetic instruments were blood-derived and the protein assays used whole hippocampal lysates, the findings should be interpreted as candidate-gene prioritization and hypothesis generation rather than proof of direct PVM-specific causality.Accordingly, the study supports a prioritized candidate framework for future functional testing, not validated therapeutic targets or direct causal proof.
This study explored how the TLR4-NF-κB-FDX1 cuproptosis axis contributes to type 1 diabetes (T1DM)-related cognitive impairment, seeking to uncover novel intervention targets. Key genes were screened from T1DM datasets via bioinformatics analysis. Primary hippocampal neurons, HBMECs, and a Transwell co-culture BBB model were established under high glucose plus copper overload. Cells received TLR4 inhibitor TAK-242 or FDX1 knockdown. A streptozotocin-induced T1DM mouse model was generated, followed by stereotaxic injection of TLR4-shRNA or FDX1-shRNA adenovirus. Cell viability, cuproptosis markers, BBB permeability, cognitive behaviors, and underlying mechanisms were assessed by CCK-8, ICP-MS, TEER, behavioral tests, Western blot, immunofluorescence, and ChIP-qPCR. FDX1 was markedly upregulated in T1DM and strongly correlated with TLR4/NF-κB. In vitro, high glucose plus copper overload activated the TLR4–NF-κB–FDX1 axis, induced DLAT lipoylation aggregation and cuproptosis, and caused neuronal synaptic injury along with disruption of endothelial tight junctions. Blocking TLR4 or knocking down FDX1 significantly attenuated these damages. In vivo, T1DM mice showed cognitive deficits and BBB disruption, both rescued by TLR4 or FDX1 knockdown. ChIP-qPCR confirmed that NF-κB bound directly to the FDX1 promoter to enhance its transcription. This study demonstrated for the first time that the TLR4–NF-κB–FDX1 cuproptosis axis plays a key pathological role in T1DM-related cognitive impairment, offering a new avenue for targeted therapy.
Hypothyroidism has been associated with cognitive disorders and high risk of neurodegenerative disorders like Alzheimer's disease (AD), which is caused by amyloid-β (Aβ) accumulation in brain. The aim of the study is to examine the therapeutic ability of triiodothyronine (T 3 ) and curcumin in regulating two key extracellular matrix proteins (ECM), collagen IV and collagen VI in brain since both drugs have neuroprotective and anti-fibrotic effects. The study further explored the possible role of signaling molecules such as transforming growth factor β1 (TGFβ1) and specificity protein 1 (Sp1) in regulating the effects of T 3 and curcumin on Aβ accumulation and ECM protein expression. Experiment was carried on Balb/c mouse model by inducing hypothyroidism with 0.05% PTU followed by treatment of T3, curcumin and a combination of both. The study revealed a significant increase in the expression of collagen IV and VI, along with elevated levels of TGFβ1 and Sp1 signalling proteins, in hypothyroid conditions in response to increased Aβ expression in both the cortex and hippocampus. Western blot and RT-PCR analysis showed that T 3 and curcumin treatment downregulate the levels of ECM proteins, the APP gene, Aβ protein, and the signaling factors. The study found that T3 and curcumin treatment downregulates the collagen IV and VI expression by negatively regulating TGFβ1 and Sp1 when expression of APP and Aβ are low. T 3 and coadministration of T 3 with curcumin more effectively regulate thyroid function and ECM protein expression in both the cortex and hippocampus. Whereas curcumin alone exhibits limited impacts under the experimental conditions in our study, this may due to bioavailability, dosage or treatment duration which need to further investigation. Our results indicate a novel therapeutic approach for understanding Aβ pathology associated with hypothyroidism, emphasizing the potential association between thyroid hormone and nature-based compounds, such as curcumin, in modulating ECM dynamics and neuroprotection.
