Search Results (126)

Mutations in the transcriptional co-factor HCFC1 cause methylmalonic aciduria and homocystinemia, cblX type (cblX) (MIM#309541), non-syndromic X-linked intellectual disability (XLID), and focal epilepsy. Zebrafish studies have revealed increased activation of the Akt/mTor signaling pathway after mutation of hcfc1a, one ortholog of HCFC1. mTOR hyperactivation is linked to seizures, and its inhibition alleviates epilepsy in other preclinical models. We hypothesized that mTor overactivity in hcfc1a mutant zebrafish increases seizure susceptibility and/or severity. We employed a two-concentration model of the seizure-inducing agent, pentylenetetrazol (PTZ), with or without pretreatment of the mTor inhibitor, torin1. Mutation of hcfc1a did not alter the response to PTZ at sub-optimal concentrations, and the pharmaceutical inhibition of mTor using the compound Torin1 reduced response to 1 µM PTZ, but only in a dose-dependent manner. Higher doses of mTor inhibition did not reduce the seizure response in mutant larvae but were effective in wildtype siblings. These data suggest that inhibition of mTor in an hcfc1a-deficient background leads to a reaction that differs from the traditional response observed in wildtype siblings. Collectively, we present a model that can be used to test dose–response and the development of combinatorial treatment approaches in a high-throughput manner. Full article

Background/Objectives: Acute seizure models are widely used in epilepsy research to investigate seizure mechanisms and evaluate antiseizure drug efficacy. Among these models, pentylenetetrazole (PTZ)-induced seizures provide a controlled and reproducible approach to study acute seizure dynamics. However, existing PTZ protocols often suffer from inconsistent seizure induction, high mortality rates, and limited translational relevance. Methods: In this study, we systematically evaluated different PTZ dosing regimens in adult Sprague-Dawley rats to establish an optimized acute seizure model that balances seizure induction efficiency with minimal lethality. Results: We tested multiple PTZ administration protocols, identifying a two-step regimen of 50 mg/kg followed by 30 mg/kg (30 min later) as the most effective strategy. This dosing approach reliably induced generalized tonic-clonic seizures in 94% of the animals while eliminating mortality. Seizures induced by this regimen were validated through electrocorticography (ECoG) recordings. The behavioral and ECoG assessments of seizures showed a strong agreement in the latency and duration (r = 0.97, p < 0.0001; r = 0.81, p < 0.05, respectively) with minimal bias in Bland–Altman analysis, confirming that both methods provide statistically comparable and interchangeable measures of seizure characteristics. Conclusions: Our findings highlight the robustness and reproducibility of this modified PTZ-induced acute seizure model, offering an improved preclinical platform for studying seizure pathophysiology and screening for novel therapeutic interventions. Full article

Objectives: One-third of patients experience inadequate seizure control with antiseizure medications. Therefore, safer and more effective therapeutic strategies remain urgently needed. Research evidence indicates that Panax notoginseng flower may exhibit potential antiepileptic properties. The study aimed to investigate the neuroprotective and antiepileptic effects of Panax notoginseng flower (PNF) extract in a chronic pentylenetetrazole (PTZ)-kindled mouse model and explore its potential mechanisms, focusing on glutamate homeostasis. Methods: Chronic epilepsy was induced in ICR mice via repeated subconvulsive PTZ intraperitoneal injections. Following successful kindling, mice were orally treated with PNF (1.5 g/kg or 3 g/kg) for 30 days. Seizure behaviors were scored using Racines scale. Neuronal survival, systemic and cerebral cytokines, hippocampal glutamate levels (in vivo microdialysis with LC-MS/MS analysis), glutamate homeostasis related proteins glutamate transporter-2 (GLT-1), glutamate-aspartate transporter-1 (GLAST), and glutamine synthetase (GS) were investigated. Results: PNF treatment significantly reduced seizure severity and restored neuronal nuclei (NeuN+) cell neurons in the cortex and hippocampal CA1 region of PTZ kindled mice. PNF attenuated systemic and neuroinflammation by lowering interleukin-1ß (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-a (TNF-α) levels and increasing interleukin-10 (IL-10) in serum and brain of PTZ mice. PNF reduced hippocampal glutamate accumulation and upregulated GLT-1, GLAST, and GS expression, which were altered by PTZ stimulation. Conclusions: The PNF extract exhibits significant neuroprotective and antiepileptic effects in PTZ-kindled mice, likely through restoring glutamate homeostasis, and suppressing inflammation. These findings, with further clinical development, support the therapeutic potential of PNF as a complementary approach for epilepsy management. Full article

Background: Epilepsy is a high-burden neurological disorder worldwide, and several sedative drugs are used as therapy. Topiramate is among the more recent drugs shown to be effective in some patients, although its benefits are limited. Two carbohydrate derivatives, FB1 (from D-fructose) and AB1 (from D-arabinose), as well as phenylboronic acid, were recently reported as sedative and safe agents in mice. Their sedative properties and structural similarity to topiramate suggest potential antiseizure activity. Objective: The objective of this study was to evaluate the antiseizure potential of FB1 and AB1. Methods: Boron-containing compounds were administered to mice with seizures induced by pentylenetetrazol (a GABA-A receptor antagonist), 4-aminopyridine (a non-selective K⁺ channel blocker), or pilocarpine (a muscarinic agonist) to assess efficacy across models and explore potential mechanisms of action. Neuronal and glial toxicity was evaluated both in vitro and in vivo. Results: AB1 reduced seizure activity after intraperitoneal administration, whereas FB1 did not exhibit anticonvulsant effects, although it modified motor performance and limited neuronal loss. The effect of AB1 was comparable to that of topiramate across all three seizure models. Docking studies suggested that these compounds can interact with GABA-A (chloride), NMDA (glutamate), calcium, and potassium channels. Toxicity assays indicated that the concentrations required to affect neurons or glial cells were ≥300 µM, supporting the safety of these compounds. Conclusions: This preliminary evaluation demonstrates the antiseizure potential of AB1. Further experimental studies are needed to clearly establish its mechanism(s) of action. Full article

Background: Epilepsy is a cluster of central nervous system (CNS) disorders identified by recurrent seizures, which affects about 60 million people around the world. In this research, a total of 40 types of 3,4,5-trimethoxycinnamic acid (TMCA) piperazine amide derivatives were designed and synthesized, inspired by the traditional Chinese medicine (TCM) herb pair drugs Polygala tenuifolia and Acori tatarinowii, followed by determination of their anticonvulsant potency. Methods: All the TMCA analogues were tested for their anticonvulsant potential through two acute models of seizures induced in mice: the maximal electroshock (MES) and sc-pentylenetetrazole (PTZ) models. In addition, the lactate dehydrogenase (LDH) inhibitory activity was determined in vitro. Results: The results showed that compounds A3, A9, A12, A14, B9, and B12 exhibited preferable anticonvulsant activity in the primary evaluation. In addition, the molecular docking results predicted good interactions of screened analogues with the LDH. Molecular dynamic simulation was used to reveal the consensual binding affinity between the most promising compound (B9) and active site interactions with LDH. Electroencephalogram (EEG) analysis and silver and immunofluorescence staining were performed to illustrate the anti-epilepsy potential of compound B9. Conclusions: Novel derivatives in this study provide new cores for the further design and optimization inspired by TCM herb pair drugs P. tenuifolia and A. tatarinowii, with the aim to explore new anticonvulsant agents. Full article

Background: The pathophysiology of epilepsy is characterized by increased neuronal activity due to an excess of the excitatory neurotransmitter glutamate and a deficiency in the inhibitory neurotransmitter gamma–aminobutyric acid (GABA). Epilepsy presents with seizures, neuronal loss, and hyperactivity in the subthalamic nucleus (STN). Astrocytes play a crucial role by absorbing extracellular glutamate through glutamate transporter–1 (GLT–1), thereby reducing neuronal excitation. Upregulating the expression of astrocytic GLT–1 is a promising therapeutic strategy for epilepsy. Sulbactam (SUL), a β–lactam antibiotic, has been demonstrated to exert neuroprotective effects by upregulating GLT–1 expression. Objectives: This study investigated the impact of SUL on neuronal and behavioral changes in epilepsy by using a pentylenetetrazol (PTZ)-induced rat model of epilepsy. Methods: Rats were treated with saline, SUL (50 and 150 mg/kg), or a combination of SUL and the GLT–1 blocker dihydrokainate (DHK) for 20 days. Subsequently, behavioral tasks were conducted to assess recognition, anxiety, and memory. Results: Histological analyses revealed that SUL ameliorated neuronal deficits, increased astrocytic GLT–1 expression, and reduced hyperactivity in the STN. Additionally, SUL promoted astrocyte proliferation, indicating a new dimension of its neuroprotective properties. However, the beneficial effects of SUL were prevented by DHK. Conclusions: This pioneering study highlights multiple benefits of SUL, including seizure suppression, increased GLT–1 expression, and astrocyte proliferation, underscoring its high potential as a treatment for epilepsy. Full article

Epilepsy remains a widespread neurological disorder, with approximately 30% of patients showing resistance to current antiepileptic therapies. To address this unmet need, a series of 2-(isoindolin-2-yl) esters derived from natural amino acids were designed and evaluated for their potential interaction with the GABA_A receptor. Sixteen derivatives were subjected to in silico assessments, including physicochemical and ADMET profiling, virtual screening–ensemble docking, and enhanced sampling molecular dynamics simulations (metadynamics calculations). Among these, compounds derived from the aromatic amino acids, phenylalanine, tyrosine, tryptophan, and histidine, exhibited superior predicted affinity, attributed to π–π stacking interactions at the benzodiazepine binding site of the GABA_A receptor. Based on computational performance, the tyrosine and tryptophan derivatives were synthesized and further assessed in vivo using the pentylenetetrazole-induced seizure model in zebrafish (Danio rerio). The tryptophan derivative produced comparable behavioral seizure reduction to the reference drug diazepam at the tested concentrations. The results implies that aromatic amino acid-derived isoindoline esters are promising anticonvulsant candidates and support the hypothesis that π–π interactions may play a critical role in modulating GABA_A receptor binding affinity. Full article

Oxidative stress and membrane damage are believed to be principally involved in the pathogenesis of epilepsy. This study aimed to assess the effects of phenosanic acid (PA), an antioxidant and membrane protector, in acute pentylenetetrazole and chronic lithium–pilocarpine seizure models in male Wistar rats. PA was administered acutely (ip, 120 mg/kg BW ip, or 240 mg/kg BW per os) or chronically (80 mg/kg BW/day per os). Indices of free radical oxidation, the hypothalamo–pituitary–adrenocortical axis, and the nitrergic system were assessed in blood and brain regions. Morphological analysis of the hippocampus was performed in the lithium–pilocarpine model. PA exerted an acute anti-seizure effect in the pentylenetetrazole model. In the lithium–pilocarpine model, acute PA treatment decreased the death rate and corticosterone levels in the neocortex and brainstem. In contrast, the level of free radical oxidation products reacting with thiobarbituric acid declined in the brain stem in response to chronic PA treatment. In the lithium–pilocarpine model, the neuronal density in the dentate gyrus was elevated, and the proliferating cell nuclear antigen positive (PCNA+) cell counts in the subgranular zone did not differ between groups. Doublecortin positive (DCX+) cell count was significantly increased after chronic PA treatment. PA-induced reduction in mortality in the lithium–pilocarpine epilepsy model may be partially mediated by decreasing the lipid peroxidation and corticosterone levels in different brain regions. Chronic PA treatment may affect adult hippocampal neurogenesis by either prolonging the action of factors that increase neurogenesis after status epilepticus or by slowing down the neuronal differentiation rate. These data suggest that PA may be a disease-modifying AED able to hamper epileptogenesis. Full article

Epilepsy is a complex neurological disorder characterized by abnormal neural synchronization and interactions between local foci and global brain networks during seizures. Understanding seizure mechanisms across multiple scales is essential for advancing our understanding of epileptic network dynamics and guiding personalized treatment strategies. However, neural recording technologies are limited by insufficient spatial resolution, signal fidelity, and the inability to simultaneously capture network- and cellular-level dynamics. To address these limitations, we developed a high-density, flexible deep-brain probe with excellent mechanical compliance and wideband recording capabilities, enabling high-fidelity recordings of high-frequency oscillations (HFOs, 80–500 Hz) and action potentials (APs). Using a pentylenetetrazol (PTZ)-induced epilepsy model, we identified distinct spatiotemporal dynamics of HFOs and APs across epileptic stages, indicating that CA3 plays a key role in seizure onset, while CA1 is crucial for propagation. AP-HFO coupling analysis further uncovered neuronal heterogeneity, offering insights into the diverse roles of neurons in epileptic networks. This study highlights the potential of a flexible deep-brain probe for advancing epilepsy research and guiding personalized therapeutic interventions. Full article

Background/Objectives: The main biologically active molecules of Cannabis sativa L. are cannabidiol (CBD) and Δ9-tetrahydrocannabinol (THC). Both exert anticonvulsant effects when evaluated as single drugs, but their possible interaction as components of C. sativa extracts has been scarcely studied. For this reason, we evaluated CBD and THC, combined or not, in two seizure models in mice, using an improved vehicle formula. Methods: Firstly, acute seizures were induced by intraperitoneal (i.p.) pentylenetetrazole (PTZ, 80 mg/kg), and mice received CBD or THC at 1, 3, 6, and 10 mg/kg, or a CBD/THC 1:1 combination at 1.5, 3, and 6 mg/kg, per os (p.o.), one hour before PTZ administration. Secondly, mice received p.o. CBD (10 mg/kg), CBD/THC (1.5, 3, and 6 mg/kg), valproic acid (50 mg/kg), or vehicle (nanoemulsions without CBD or THC), one hour before PTZ (30 mg/kg, i.p.) every other day for 21 days. Behavioral, biochemical, and immunohistochemical analyses were performed to assess the response to PTZ, oxidative stress, and astroglial activation. Results: In the acute model, CBD and THC at 3–10 mg/kg, and their combinations, significantly increased latency to generalized seizures and death, and improved survival rates. In the chronic model, similarly to valproic acid, CBD 10 mg/kg and CBD/THC at 1.5 and 3 mg/kg delayed kindling acquisition, while CBD/THC 6 mg/kg had no effect. CBD and CBD/THC treatments reduced oxidative and nitrosative stress and attenuated astrogliosis, as indicated by decreased glial fibrillary acidic protein and GABA transporter 1 expression and increased inwardly rectifying potassium channel 4.1 expression in hippocampal regions. However, no cannabinoid treatment prevented the impairment in novel object recognition and Y maze tests. Conclusions: These findings support the potential role of cannabinoids in counteracting seizures, possibly by reducing oxidative stress and astrogliosis. The study also highlights the importance of nanoemulsions as a delivery vehicle to enhance cannabinoid effectiveness while considering the risks associated with direct cannabinoid receptor activation. Full article

Background/Objectives: Epilepsy is a common neurological condition characterized by the occurrence of a seizure. It affects around 50 million individuals worldwide, and despite the large quantity of anti-seizure medications available, 30% of epileptic patients still suffer from seizures. Therefore, it is necessary to find new therapeutic options. Interestingly, polydatin has shown promising effects on epilepsy treatment due to its antioxidant and anti-inflammatory properties. Thus, this study aimed to evaluate the effects of polydatin (200, 300, and 400 µM) on a pentylenetetrazol (PTZ)-induced seizure model in wild-type zebrafish (Danio rerio) larvae. Methods: Seizure-like behavior, cell death, reactive species (RS) production, and lipid peroxidation were analyzed. Results: Pre-treatment with polydatin at 200 and 300 µM did not have a significant impact on seizure occurrence and the behavior of animals exposed to PTZ. Diazepam decreased seizure occurrence and increased the latency to achieve each seizure stage. Exposure to PTZ increased the swimming activity, and this effect was suppressed by diazepam but not by polydatin. PTZ exposure increased the RS production, which was significantly attenuated by polydatin at 400 µM and DMSO. Cell death and lipid peroxidation were not changed when compared to the experimental groups. Conclusions: Only the experimental positive control (diazepam) showed anti-seizure effects. Therefore, we failed to observe any anti-seizure effects of polydatin using a zebrafish experimental model. However, we cannot rule out its effects in other experimental models and different treatment protocols. Full article

Background/Objectives: Hemorphins, considered to be bioactive atypical oligopeptides, are products of hemoglobin metabolism. Recently, our team reported the synthesis and characterization of three N-modified analogs of hemorphin-4 (H4) with rhodamine B (Rh). In the present study, the Rh-1, Rh-2, and Rh-3 compounds were intracerebroventricularly infused at doses of 1, 2.5, 5, and 10 µg/5 µL, respectively, and evaluated for their antiseizure activity in 6-Hz and maximal electroshock (MES) tests and in a pentylenetetrazol (PTZ)-induced kindling model in mice. Phenytoin and diazepam were used as the reference drugs. The role of opioid receptors (ORs) underlying their mechanism of action was also evaluated in silico and pharmacologically. Results: The three Rh-H4 compounds showed a good safety profile at a concentration of 100 µg/mL in the mouse embryonic fibroblasts. They suppressed psychomotor seizures and seizure spreading as follows: Rh-1 at doses of 5 and 10 µg/5 µL, Rh-2 at the highest dose, and Rh-3 at doses of 1–10 µg/5 µL, respectively. Administered at doses of 5 µg/5 µL (Rh-1 and Rh-3) and 10 µg/5 µL (Rh-2), the compounds suppressed clonic seizures in the kindled mice comparable to the reference drug diazepam. A combination of selective delta (DOR), kappa (KOR), and mu (MOR) OR antagonists with the highest doses of the Rh-1, Rh-2, and Rh-3 compounds was used to elucidate the possible role of ORs in the underlying mechanism related to their protective activity against seizure spread. Only the selective DOR antagonist, natrindole, suppressed the effect of the Rh-1 peptide analog on seizures. The OR antagonist naloxone prevented the antiseizure activity of Rh-1 in the kindled mice. The results of docking analysis also showed the model-specific interaction of the three Rh-H4 compounds with the OR. Conclusions: Our results suggest that the antiseizure activity of Rh-1 is mediated by the OR, and in particular by the DOR, while the mechanism underlying the antiseizure effect of Rh-3 is more complex and may involve other receptors. Full article

Epilepsy is a widespread neurological disorder that remains a critical global public health challenge. While numerous antiepileptic drugs (AEDs) are available, many patients either fail to achieve adequate seizure control or experience significant side effects. One promising alternative is pure cannabidiol (CBD), but using a whole cannabis extract may be equally effective and preferred for some patients. In the current study, we employed the pentylenetetrazole (PTZ)-induced hyperactivity model in zebrafish to compare the effects of CBD with various cannabis extracts. We evaluated three cannabis strains, each subjected to three different extraction methods, and benchmarked the results against the commercially available AED valproic acid (VPA). Our findings revealed that 5.7 µg/mL of CBD and 10 µg/mL of different extracts significantly reduced movement compared to PTZ and VPA. In addition, effective extracts produced effects similar to pure CBD despite containing much lower molecule levels. These results reinforced and expanded previous evidence supporting the clinical potential of both CBD and whole cannabis extracts for seizure control while suggesting a possible entourage effect. Further research is necessary to determine which patients may benefit more from pure CBD versus those who might prefer whole cannabis extracts. Full article

As a phytotherapeutic agent with historical applications in epilepsy management, Acorus tatarinowii Schott (ATS) remains pharmacologically enigmatic, particularly regarding its pathophysiological mechanisms. This knowledge gap significantly hinders the clinical application of ATS-based treatments. To explore the potential of ATS in combating epileptogenesis, we utilized a pentylenetetrazole (PTZ)-induced chronic epilepsy rat model. Brain metabolomic analysis was performed by ultra-performance liquid chromatography coupled with mass spectrometry (UPLC/MS). Principal component analysis (PCA) and orthogonal projections to latent structures-discriminant analysis (OPLS-DA) were performed for screening differential metabolites. Gut microbiota composition was analyzed through 16S rRNA gene sequencing and examined using Spearman correlation analysis. The results show that oral ATS (50 mg/kg) significantly improved the seizure latency and pathology of rats with epilepsy. Ascorbate and aldarate metabolism, glycerophospholipid metabolism, arachidonic acid metabolism, and intestinal flora were crucial for ATS’s ability to counteract epilepsy. The therapeutic effects of ATS against epilepsy were investigated with brain metabolomics and gut microbiota analysis, providing the basis for further comprehensive research. Full article

Zuranolone (SAGE-217), the first FDA-approved oral neurosteroid (NAS), a positive allosteric modulator (PAM) of γ-aminobutyric acid type A (GABA_A) receptor for postpartum depression approved in 2023, has limitations such as short half-life, low bioavailability, and central inhibitory side effects. To address these, we designed novel C-21 modified derivatives of Zuranolone, identifying the triazolone scaffold as key for enhancing GABA_A activity. Here, we synthesized Zuranolone analogs with diverse triazolone substituents, finding that pyridine-derived modifications improved the activity correlated with LogP. The optimal derivative, S9 (2-(trifluoroethoxy)pyridine-triazolone, LogP 4.61), showed 2.5-fold greater potency (EC₅₀) and efficacy (Emax) than Zuranolone (LogP 4.78) at synaptic/extrasynaptic GABA_A receptors, attributed to stronger binding via molecular docking. In rats, S9 exhibited 5-fold longer plasma T_1/2, 6-fold higher AUC, 3-fold greater brain exposure, and 30% improved bioavailability. It also outperformed Zuranolone in pentylenetetrazole (PTZ)-induced seizure suppression and threshold dose for loss of righting reflex (LORR) in rats. The C21-pyridine-triazolone pharmacophore in S9 enhances receptor activity potency without increasing lipophilicity, optimizing pharmacokinetics and safety, which makes it a promising therapeutic candidate for depression and epilepsy. Full article