Alzheimer’s disease (AD) is an increasingly prevalent neurodegenerative disorder worldwide, with women showing a higher risk of developing the disease. The decline in steroid hormones after menopause may contribute to the increased susceptibility of women to neurodegenerative conditions. In parallel, cannabis-derived compounds have been reported to alleviate certain symptoms associated with neurological disorders. The present study was designed to investigate the effects of marijuana extract on cognitive impairment and hippocampal molecular markers in an ovariectomized AD-like rat model, and to evaluate whether co-administration with estradiol modifies these effects. The marijuana extract used in this study was characterized by HPLC and was found to contain 8.5% Δ9-THC. AD-like cognitive impairment pathology was induced by intra-hippocampal administration of Aβ25–35 in OVX rats. Animals were treated with marijuana extract (60mg/kg/day, corresponding to approximately 5.1mg/kg/day Δ9-THC) either alone or in combination with 17β-estradiol (1mg/kg every 4 days) for 28 days. Cognitive performance was evaluated using the Morris water maze (MWM). In addition, hippocampal CB1/CB2 receptor expression and BDNF protein levels were measured to examine potential molecular associations. Our findings showed that chronic administration of the marijuana extract improved Aβ25–35-induced deficits in spatial learning and memory. Alterations in CB1 receptor expression and BDNF levels accompanied these behavioral effects. Notably, co-treatment with estradiol did not produce a synergistic effect, suggesting a complex interaction between cannabinoid-related and estrogen-related signaling pathways. These preclinical findings suggest that a THC-standardized marijuana extract may exert neuroprotective-like effects in an AD-like cognitive impairment model. However, because the extract was not fully phytochemically characterized and the mechanistic analyses were correlational, the results should be interpreted cautiously. Further studies are needed to clarify the underlying mechanisms and translational relevance of these findings.
Our previous study demonstrated that ovariectomy exacerbated stress-induced impairments in rats exposed to single prolonged stress (SPS) and that the treadmill exercise protocol used in that study improved the associated cellular responses. However, this protocol did not alleviate all anxiety-like behaviors. Therefore, the present study aimed to determine whether the voluntary wheel-running protocol used in this study would produce greater improvements in anxiety-like behavior than the previously evaluated treadmill exercise protocol.Female adult Wistar rats were randomly allocated to the control (CON), sham, and ovariectomized (OVX) groups. Following PTSD induction, animals within each group were randomly assigned to sedentary (SED), treadmill running (TR), or wheel running (RW) groups. Twenty days after ovariectomy, PTSD was induced by a Single Prolonged Stress (SPS) model. The exercise began on day 32 and continued for 4 weeks. On the 61st day, Open Field Test (OFT), Object Location Memory Task (OLMT), corticosterone, and neurotrophins levels were assessed. p<0.05 was considered a significant level.The findings revealed significant differences between the OVX/SPS/TR and OVX/SPS/RW groups in hippocampal and prefrontal cortical BDNF and IGF-1 levels. Significant differences were also observed between the OVX/SPS/SED group and both the OVX/SPS/TR and OVX/SPS/RW groups in the OLMT and OFT.The wheel-running protocol produced greater therapeutic benefits against SPS-induced impairments that were exacerbated by ovariectomy. Based on the results of the present study, the exercise protocol appears to be an important factor in achieving targeted therapeutic outcomes in an animal model of PTSD. Exercise-induced stress load, as well as dose and intensity should be considered when drawing conclusions.
Introduction Dimethyl fumarate (DMF) is an oral disease-modifying therapy recommended for patients with relapsing-remitting multiple sclerosis (MS). However, a lack of efficacy, safety concerns, tolerability issues, and poor compliance often lead to discontinuation of DMF. In an innovative approach, this exploratory proof-of-concept clinical trial aimed to assess the feasibility and safety of introducing variability into treatment regimens to overcome compensatory mechanisms that underlie the loss of drug effectiveness and to generate preliminary signals of potential improvements in response to DMF. Methods Seven MS patients treated with DMF were enrolled in an open-label, uncontrolled feasibility clinical trial in which an app provided a personalized therapeutic regimen, resulting in variability in dosages and administration times within predefined ranges. Prespecified feasibility endpoints included: (1) successful app installation and use; (2) patient adherence to the algorithmic dosing schedule; (3) completion of the 12-week follow-up; and (4) absence of serious adverse events. Exploratory outcome measures included the Expanded Disability Status Scale (EDSS), neurofilament light chain (NfL) levels, and MRI, which were assessed only as hypothesis-generating signals. Results Although this study is a small exploratory proof of concept, the data suggest that AI-assisted personalized treatment is feasible and could improve clinical responses to DMF. The EDSS score improved in 2 patients (29%, p = 0.371), and the MRI remained stable in 6 patients (100%). These findings did not reach statistical significance and should be interpreted as exploratory only. A high patient engagement rate with the app was recorded during the study. No serious adverse events were observed. Summary The results of this feasibility trial support the safety and feasibility of using a personalized algorithm that randomizes DMF regimens and provide preliminary hypothesis-generating signal data regarding potential improvements in therapy response. These findings are insufficient to support conclusions about efficacy. Controlled studies are needed to confirm these findings. (NCT06385197)
Background:Older adults with Alzheimer's disease (AD) frequently present to emergency departments (EDs) with multiple coexisting conditions. However, age-specific patterns of multimorbidity in AD-related ED encounters remain incompletely characterized. Objective:To identify and describe age-specific multimorbidity subtypes among AD-associated ED encounters using unsupervised machine learning and heatmap-based diagnostic profiling. Methods:We conducted a cross-sectional analysis of the 2022 Nationwide Emergency Department Sample. AD-associated encounters were identified by an ICD-10-CM G30.x diagnosis code in any diagnosis field. The cohort included 125,461 ED encounters and was stratified into four age groups: 60-64, 65-74, 75-84, and ≥ 85 years. For each age group, the 30 most frequent co-occurring diagnoses were converted into binary indicators. KMeans clustering with eight clusters per age group was used for heatmap-based subtyping, while Uniform Manifold Approximation and Projection and Hierarchical Density-Based Spatial Clustering of Applications with Noise were used to visualize diagnostic structure. Results:Distinct multimorbidity profiles were identified across all age groups. Among adults aged 60-74 years, clusters commonly included psychiatric and metabolic conditions, such as depression, anxiety, diabetes, chronic kidney disease, substance-use diagnoses, and dehydration. Among adults aged ≥ 75 years, clusters more frequently included cardiorenal disease, urinary tract infection, metabolic encephalopathy, respiratory failure, do-not-resuscitate status, and palliative care. The cohort had a mean age of 81.7 ± 7.2 years, and 62.6% of encounters involved women. Conclusion:Patterns of coexisting conditions in AD-related ED visits differed substantially by age. Adults aged 60-74 years more often showed psychiatric and metabolic combinations, whereas those aged ≥ 75 years more often showed infection, cardiorenal disease, respiratory failure, and end-of-life care markers. Recognizing these age-related patterns may help clinicians anticipate common care needs when evaluating patients with AD in the ED.
Type 1 diabetes mellitus is a chronic disease where the destruction of insulin-producing pancreatic β cells causes hyperglycemia and its onset typically occurs during childhood or adolescence. We previously reported that long-term depression (LTD) in the CA1 region of the hippocampus, induced by a low-frequency stimulation to the Schaffer collaterals, was attenuated in juvenile-onset diabetes (JDM) rats. We herein investigated whether LTD was triggered by the activation of G-protein coupled group I metabotropic glutamate receptors (mGluR-LTD) or M1 muscarinic acetylcholine receptors (mAChR-LTD) at Schaffer collateral-CA1 synapses in hippocampal slices from JDM rats. Our results indicated that the magnitude of mGluR-LTD induced by the group I mGluRs agonist (S)-3,5-dihydroxyphenylglycine (DHPG) was smaller in JDM rats than in age-matched control rats. Paired-pulse facilitation (PPF) is primarily associated with enhanced presynaptic transmitter release. To investigate the interaction between PPF and mGluR-LTD, PPF ratios conducted at interstimulus intervals (ISIs) of 50 and 200 ms were compared at the baseline and LTD stage. In control rats, the PPF ratio at ISIs of 50 ms was higher during the mGluR-LTD stage than at the baseline. However, PPF ratios at ISIs of 50 and 200 ms were higher during the mGluR-LTD stage than at the baseline in JDM rats. In contrast, the magnitude of mAChR-LTD did not significantly differ between control and JDM rats. Therefore, the expression of mGluR-LTD in JDM rats may be associated with presynaptic changes, since the increase in the PPF ratio occurred concomitantly with the attenuation of LTD.
Alzheimer's disease (AD) involves complex changes, including synaptic dysfunction, neuroinflammation, and metabolic impairment, yet the role of lipid raft-associated gene networks in these processes remains unclear. In this study, we performed an integrative transcriptomic analysis using the GSE5281 dataset and validated the findings in an independent cohort (GSE33000). By combining differential expression analysis with weighted gene co-expression network analysis, we identified 72 lipid raft-associated genes linked to mitochondrial, synaptic, and immune-related pathways. Using network analysis and machine learning approaches (LASSO and SHAP), we further identified eight key biomarkers (CBL, CD44, EZR, FAS, FYN, ITGB1, MAPK1, and TGFBR1) that showed strong diagnostic performance and consistent results across datasets. Functional analysis revealed increased inflammatory signaling, including pyroptosis and cytokine pathways, alongside reduced oxidative phosphorylation and synaptic activity. Interestingly, immune profiling showed only minor differences in immune cell infiltration, but clear activation of multiple immune pathways, such as Th2 and Treg signaling, CD8⁺ T-cell signatures, and IL-6/IL-10-mediated inflammation. These immune changes were strongly associated with lipid raft-related biomarkers. Overall, our findings suggest that lipid raft dysregulation may act as a key link between immune activation, mitochondrial dysfunction, and synaptic impairment in AD.
Background Post-traumatic stress disorder (PTSD) arises after traumatic or life-threatening events and involves persistent anxiety and altered stress responses. Physical exercise is a proposed non-pharmacological intervention, yet evidence comparing voluntary and forced exercise regarding their behavioral and biochemical effects in experimental PTSD models is still limited. Methods In this experimental study, adult male rats were subjected to a single prolonged stress (SPS) protocol to induce a PTSD-like phenotype and were subsequently assigned to voluntary exercise, forced exercise, or control conditions. Anxiety-like behaviors were evaluated using behavioral tests, including the open field test, elevated plus maze, and light–dark box. The hippocampal mRNA expression levels of inflammatory cytokines, including IL-1β, IL‑10, and TNF‑α, were quantified by reverse transcription polymerase chain reaction. Results Exposure to SPS induced a marked anxiety‑like behavioral phenotype, as evidenced by reduced exploration of anxiogenic zones across all behavioral paradigms, without significantly affecting general locomotor activity. SPS exposure was also associated with increased hippocampal expression of the pro‑inflammatory cytokines IL‑1β and TNF‑α, accompanied by reduced expression of the anti‑inflammatory cytokine IL‑10. Both voluntary and forced exercise attenuated SPS-induced anxiety-like behaviors and significantly improved hippocampal inflammatory gene expression. Voluntary exercise produced greater improvement across several behavioral measures. At the molecular level, both exercise modalities attenuated SPS-induced pro-inflammatory alterations, whereas voluntary exercise alone significantly restored the anti-inflammatory cytokine IL-10. Conclusion Both voluntary and forced exercise effectively attenuated SPS-induced anxiety-like behavior and hippocampal inflammatory dysregulation. Voluntary exercise demonstrated greater behavioral improvement across anxiety-related paradigms. Although only the VolExc group reached statistical significance relative to the SPS group for IL-10, direct comparison between VolExc and ForExc did not reveal a significant difference, indicating that molecular outcomes were broadly comparable between the two exercise modalities. These findings suggest that voluntary exercise may provide behavioral advantages following traumatic stress, although the two modalities were not statistically distinguishable from one another at the molecular level.
AD is a neurodegenerative disorder marked by progressive cognitive decline, particularly memory impairment, largely driven by cholinergic dysfunction. This study aimed to investigate the pharmacological basis and modes of action of the identified TT-TeMac™ compounds againsts cholinergic dysfunction associated with memory loss. TT-TeMac™ compounds targets which were identified by LC-MS were extracted from SwissTarget Prediction and PharmMapper, while cholinergic dysfunction-related targets were obtained from DisGeNET and GeneCards. Fifteen common targets were identified, with nine key targets highlighted by STRING, Cytoscape, and Venny analyses and confirmed by molecular docking using MOE. The in vivo validation used a rat model with scopolamine-induced cholinergic dysfunction (1 mg/kg bw/day, ip) for 7 days. Subsequently, behavioral (NOR and MWM), biochemical (AchE and BuhE) and histological (H&E and CV) analyses were performed. Seven compounds were identified in TT-TeMac™ (terminolic acid, sericic acid, arjunolic acid, gallic acid, ellagic acid, 3-O-methyl ellagic acid and 3,3'-di-O-methyl ellagic acid). Network pharmacological analysis showed that TT-TeMac™ acted on 15 common targets of which ACHE, IL6, TNF, SNCA, AKT1, SERPINE1, STAT3, ACE, and ALB were the pivotal genes. Also, docking studies confirmed the involvement of the target within the network with meaningful binding energies. Furthermore, TT-TeMac™ prevented cholinergic dysfunction associated with memory loss in rats by significantly reducing cholinesterase activity and protecting against morphological alterations and neuronal loss in the hippocampus. Our study shows that the ingredient TT-TeMac™ has a multi-targeted mode of action on protein targets involved in cholinergic dysfunction and counteracts this dysfunction in scopolamine-treated rats.
Aging and Alzheimer's disease are complex processes marked by continuous neuronal loss, disrupted neural networks, and cognitive decline. The past decade has seen advances in genomics, proteomics, and single-cell RNA sequencing, enabling the discovery of cellular markers with distinct gene expression patterns. This review aims to compile recent markers of aging and Alzheimer’s disease in immune, glial, and neuronal cells, which are increasingly vital for enhancing diagnostic precision and monitoring disease progression. Markers such as Triggering receptor expressed on myeloid cells 2 (TREM2) and Cluster of differentiation 33 (CD33) facilitate immune responses and function as indicators of neuroinflammation and amyloid-beta clearance; Glial fibrillary acidic protein (GFAP) and aquaporin-4 serve as indicators of gliosis and impaired interstitial fluid drainage; Postsynaptic density protein 95 (PSD-95), Synaptosomal-associated protein 25 (SNAP25), and aberrant tau phosphorylation signify synaptic degradation and cytoskeletal instability characteristic of Alzheimer's pathology. The breakdown of the blood-brain barrier is associated with endothelial nitric oxide synthase (eNOS) and vascular cell adhesion molecule 1 (VCAM-1). The markers were identified using cutting-edge technologies that unveiled variations in gene expression across cell types, brain regions, and disease stages. This cellular heterogeneity improves understanding of Alzheimer's disease (AD) progression and brain aging, clarifies molecular pathways, and may be used for prognostic and diagnostic purposes after thorough validation. Novel cellular markers with altered expression profiles are still being identified through ongoing research, which could be crucial for improving our understanding of and ability to treat neurodegenerative diseases.
Nuclear factor kappa B (NF-κB) has been implicated in both protective and detrimental processes in Alzheimer disease (AD), creating an apparent contradiction in the literature. Increasing evidence, however, indicates that these divergent effects reflect differences in cell type, molecular configuration, activation kinetics, disease state, and the surrounding pathological environment rather than a true biological paradox. Under transient and tightly regulated conditions, NF-κB can support neuronal survival, stress adaptation, antioxidant defense, and selected compensatory responses to amyloid-β pathology. In contrast, persistent or disease-conditioned signaling is more consistently associated with amyloidogenic processing, tau propagation, chronic neuroinflammation, impaired proteostasis, neuronal dysfunction, and neurovascular injury. The available evidence therefore supports an asymmetric framework in which protective effects are confined to relatively specific molecular and temporal settings, whereas sustained pathological signaling is supported across a broader range of disease-relevant models and cellular processes. This distinction argues against indiscriminate activation or global inhibition of NF-κB, either of which could disrupt physiological functions while failing to selectively suppress disease-driving pathways. Therapeutic strategies should instead target defined pathological NF-κB programs within the relevant cell type and disease context while preserving adaptive and homeostatic signaling. This review synthesizes the context-dependent roles of NF-κB in AD and provides a framework for reconciling its apparently opposing effects.
The "Xingnao Kaiqiao" method is an acupuncture approach primarily applied at the PC6 and GV26 acupoints for ischemic stroke (IS). Whether Electroacupuncture (EA) at these acupoints influences the cGAS-STING signaling pathway, affects microglial polarization, and reduces neuroinflammation in IS was the aim of this study. Adult male Sprague-Dawley rats were randomly assigned to five groups: sham, middle cerebral artery occlusion (MCAO), MCAO + EA, MCAO + RU.521, and MCAO + EA + cGAMP. Neurological function scoring, 2,3,5-triphenyltetrazolium chloride (TTC) staining, hematoxylin-eosin (HE) staining, Nissl staining, Masson staining, flow cytometry, enzyme-linked immunosorbent assay (ELISA), RT-qPCR, and Western blotting were conducted for comprehensive evaluation. The MCAO group showed significant neurological impairments, widespread brain damage, and showed elevated pro-inflammatory cytokine levels relative to sham controls. These pathological alterations were substantially reversed by EA treatment. EA administered at the PC6 and GV26 acupoints suppresses excessive activation of the cGAS-STING signaling cascade, thereby decreasing the production of pro-inflammatory mediators while enhancing anti-inflammatory responses. This modulation further facilitates the shift of microglia (MG) from a pro-inflammatory M1 state toward a reparative M2 phenotype. These changes collectively alleviate neuroinflammation and mitigate brain injury.
Sleep disorders such as insomnia, sleep apnea, and circadian rhythm dysfunctions constitute a major public health challenge due to their high prevalence and associated risks for physical and mental health. In response to the limitations and potential side effects of pharmacological treatments, non-pharmacological interventions have become increasingly central to management strategies. Among these, structured physical exercise has emerged as a particularly prominent and evidence-supported approach, valued for its demonstrated efficacy and favorable safety profile. This narrative review aims to consolidate and examine the contemporary body of scientific evidence elucidating how distinct forms of exercise, primarily categorized as aerobic endurance training, resistance or strength training, and mindful mind-body practices, act to alleviate various sleep disturbances. The underlying biological pathways through which these exercise modalities exert their positive effects are complex and interconnected, encompassing thermoregulatory processes, neurochemical adaptations, endocrine signaling, and shifts in autonomic nervous system balance.
Background Topiramate (TPM) is a broad-spectrum antiseizure medication associated with dose-limited mood-related adverse effects, including depression, which significantly impair patient quality of life and treatment adherence. Although physical exercise confers neuroprotective and mood-stabilizing benefits, its potential to counteract TPM-induced behavioral deficits and the underlying neurobiological mechanisms, particularly within the serotonergic system, remains largely unexplored. Objective The present study investigated whether exercise reduced the effective dose of topiramate and attenuates its depressive-like side effects in epileptic rats. Methods Male Wistar rats were randomly assigned to nine groups (n = 10/group): Sham, Seizure, Exercise (EX), TPM (25 mg), TPM (50 mg), TPM (70 mg), EX + TPM (25 mg), EX + TPM (50 mg), and EX + TPM (70 mg).The exercise protocol consisted of treadmill running (30 min/day, 5 days/week for 4 weeks). Seizure severity and latency were assessed using the Racine scale. Depressive-like behaviors and anxiety were evaluated by the Tail Suspension Test (TST) and the Elevated Plus Maze (EPM). Immunohistochemistry was performed to quantify 5-HT1A receptor immunoreactive staining intensity in the hippocampal CA1, CA3, and cerebral cortex. Results Combination therapy (EX+TPM (50 mg) and EX + TPM (70 mg)) demonstrated superior anticonvulsant effects, significantly reducing seizure scores and prolonging seizure latency compared to TPM monotherapy (p < 0.001). TPM monotherapy dose-dependently exacerbated depressive-like behaviors (e.g., increased immobility in the TPM (70 mg) group, p < 0.001). Crucially, exercise attenuated these adverse effects in all EX + TPM groups. 5-HT1A receptor expression was significantly downregulated in the Seizure and TPM (25 mg) groups compared with the Sham and Exercise groups. In contrast, the EX + TPM (50 mg) and EX + TPM (70 mg) groups showed significant upregulation of 5-HT1A receptors in the hippocampus and cerebral cortex (p < 0.001). Conclusion The current findings demonstrate that adjunctive moderate-intensity treadmill exercise potentiates the anticonvulsant efficacy of topiramate and effectively counteracts its depressive-like side effects and anxiety. This behavioral improvement was correlated with exercise-mediated upregulation of 5-HT1A receptors, suggesting a potential mechanistic link. These findings provide a preclinical rationale for further investigation into the potential benefits of exercise as an adjunctive therapy in epilepsy, though clinical translation requires additional studies.
Background:This study aimed to comprehensively examine the interplay between demographic factors, peripheral oxidative stress markers, disability, depressive symptoms, cognitive outcomes, and hand dexterity in Relapsing-remitting Multiple sclerosis (RRMS) patients with mild/moderate disability. Methods:In this cross-sectional study, RRMS patients with Expanded Disability Status Scale (EDSS)≤ 3 underwent assessments of cognitive function (Symbol Digit Modalities Test [SDMT], Paced Auditory Serial Addition Test [PASAT-3]), depressive symptoms (Beck Depression Inventory-II [BDI-II]), hand dexterity (Nine-Hole Peg Test [9HPT]), and neurological disability (EDSS). Oxidative stress markers (malondialdehyde [MDA], total antioxidant capacity [TAC], superoxide dismutase [SOD], catalase activity [CAT], glutathione peroxidase [GPx], and reduced glutathione [GSH]) were also measured. Results:The studied cohort comprised 83 patients (77.1% females; mean age: 32.65 ± 8.05 years). In addition to negative association between the EDSS and cognitive outcomes in binary logistic regression analysis (SDMT: OR=0.95; p = 0.02; PASAT: OR=0.92; p = 0.01); neurological disability also correlated with cognitive scores in multivariable linear regression (SDMT: β=-0.23, p = 0.047; PASAT: β=-0.36, p = 0.002). Education also correlated with the cognitive outcomes (SDMT: β=0.29, p < 0.001; PASAT: β=0.27, p = 0.01). MDA was associated with abnormal cognitive function (SDMT≤44; OR: 2.98 (95%CI: 1.33, 6.66) but correlation between these outcomes was no longer significant in multivariable linear regression (β=-0.21; p = 0.051). Except for a marginal correlation between EDSS and MDA (β=0.21, p = 0.04) and between 9HPT-M and TAC (β=0.26, p = 0.01), there was no significant finding regarding the correlations between the clinical and laboratory outcomes. Conclusion:Even in patients with mild/moderate disability, EDSS and education level were emerged as the strongest predictors of cognitive outcomes in RRMS.
Objective:This study aimed to investigate the association between the HOTAIR-miR-9-5p axis and the inflammatory response in ischemic stroke (IS) and elucidate the underlying molecular mechanisms. Methods:Middle cerebral artery occlusion/reperfusion (MCAO/R) and oxygen-glucose deprivation/reoxygenation (OGD/R) were applied to simulate ischemic/reperfusion conditions in vivo and in vitro. The expression levels of HOTAIR and miR-9-5p in the serum of patients or in the brain tissue of MCAO/R mice were assessed by qRT-PCR, and the secretion levels of IL-1β and IL-18 were analyzed by ELISA. Dual-luciferase reporter assays, RNA-binding protein immunoprecipitation (RIP), and RNA pull-down assays were performed to validate the target relationship. Western blotting was applied to assess the expression of NLRP3, CASP1, and FOXP1. Moreover, MCAO/R mice with intracerebroventricular injection of antagomir-9-5p were used to evaluate the effect of antagomir-9-5p on cerebral ischemia-reperfusion injury (CIRI). Results:A significant association was observed between the HOTAIR-miR-9-5p axis and inflammation in both IS patients and MCAO/R mice. HOTAIR was abnormally expressed at a low level, whereas miR-9-5p and the associated protein NLRP3 inflammatory response were increased in the serum of IS patients, as well as in the brain tissue of MCAO/R mice and in SH-SY5Y cells. Mechanistically, miR-9-5p negatively regulated the expression of HOTAIR and FOXP1. Furthermore, HOTAIR was found to regulate NLRP3 expression via the miR-9-5p/FOXP1 pathway. Functional experiments revealed that silencing miR-9-5p protected against cerebral ischemia/reperfusion injury and suppressed NLRP3 inflammasome activation. Conclusion:These findings collectively demonstrate that the HOTAIR/miR-9-5p/FOXP1 axis plays a critical role in NLRP3 inflammasome activation following IS, suggesting that its blockade could be a potential therapeutic strategy for ischemic brain injury.
Purpose The ketogenic diet (KD) is a well-established therapy for drug-resistant epilepsy, reducing seizure burden in children and improving cognition in epileptic encephalopathy patients, but its underlying mechanisms remain incompletely understood. This study aimed to establish a translational zebrafish model to investigate KD’s anti-epileptic potential and mechanisms, focusing on neurotransmitter homeostasis. Method Acute epilepsy was induced in 7-day post-fertilization (dpf) zebrafish larvae (n = 24/group, randomly assigned) using pentylenetetrazol (PTZ), a selective GABA-A receptor antagonist. KD was administered to the treatment group, with behavioral assays (locomotor activity, thigmotaxis), electroencephalographic (EEG) recordings, and high-performance liquid chromatography (HPLC) for neurotransmitter quantification. Statistics were analyzed via two-way ANOVA/Tukey’s test or Student’s t-test. Results Results showed KD significantly ameliorated PTZ-induced epileptiform behaviors (prolonged seizure latency, reduced stage-specific duration), attenuated EEG multi-peak discharges, increased inhibitory neurotransmitters (GABA, glycine, taurine), and reversed PTZ-induced excitatory-inhibitory neurotransmitter imbalance (e.g., Glu/GABA, Gln/Tau ratios). Conclusion We established a PTZ-induced amphibian epilepsy model with KD intervention. KD exerts anti-epileptic effects, likely mediated partially by rebalancing central nervous system neurotransmitter homeostasis, providing a valuable translational platform for further mechanism research.
Microglia-mediated inflammation plays a critical role in secondary brain injury following ischemic stroke. Glycogen synthase kinase-3β (GSK-3β) and hexokinase 2 (HK2) are involved in regulating microglial inflammatory responses and can promote the release of pro-inflammatory factors under ischemic and hypoxic conditions, thereby exacerbating brain damage. Early exercise intervention has been shown to improve post-stroke neuroinflammation, but whether it exerts this effect by modulating GSK-3β/HK2-associated inflammatory signaling in microglia remains unclear. This study aimed to investigate the effects of early exercise on microglial inflammation and the potential involvement of GSK-3β/HK2 signaling. A twenty-eight-day early exercise regimen was applied in a rat model of middle cerebral artery occlusion (MCAO), and complementary in vitro experiments were conducted using BV2 microglial cells under oxygen-glucose deprivation. Results demonstrated that early exercise significantly improved neurological outcomes and reduced brain tissue damage, accompanied by decreased expression of microglial pro-inflammatory factors (IL-6, TNF-α, CD86, and iNOS) and increased levels of anti-inflammatory factors (IL-10 and Arg1). In vitro, inhibition of GSK-3β in BV2 cells markedly alleviated inflammatory responses under hypoxic conditions. In conclusion, early exercise intervention can attenuate post-stroke neuroinflammation, potentially by modulating GSK-3β/HK2 signaling in microglia to regulate.
