Hippocampus Of Epileptic Rats Research Articles

Exploring the Therapeutic Potential of Baicalin: Mitigating Anxiety and Depression in Epileptic Rats.

Epilepsy is a serious neurological disorder that affects millions of people each year, often leading to cognitive issues and reduced quality of life. Medication is the main treatment, but many patients experience negative side effects. Male Sprague-Dawley (SD) rats were chosen as experimental animals for this experiment due to their physiological and genetic similarities to humans, cost-effectiveness, and ease of handling in a laboratory setting. The objective of this study was to assess the neuroprotective properties of baicalin (BA) in relation to its impact on anxiety and depressive-like behaviors in the epilepsy model. Thirty male Sprague-Dawley (SD) rats were selected for this experiment. Pentylenetetrazol (PTZ) kindling (40 mg/kg; i.p.) was utilized to establish an epilepsy model. The effect of BA (50 mg/kg; gavage) on seizure severity (assessed using the Racine scale), anxiety, and depressive- like behaviors (evaluated through open field experiments and forced swimming tests) was examined. Histological examinations, including hematoxylin and eosin (HE) staining and Nissl staining, were conducted to assess neuronal damage. Furthermore, the neuroprotective properties of BA were examined through the analysis of Doublecortin (DCX), MKI67 (KI67), and Brain-Derived Neurotrophic Factor (BDNF) levels in the hippocampus of rats. The inhibitory impact of BA on neuroinflammation was assessed via dual labeling for NOD-like receptor thermal protein domain associated protein 3 (NLRP3) and the microglial marker ionized calcium- binding adapter molecule 1 (Iba-1). The influence of BA on the expression of P2X7 receptor (P2X7R), NLRP3, and Interleukin-1β (IL-1β) was also assessed by reverse transcription quantitative PCR (RT-qPCR) in the brain. Finally, we employed a molecular docking model to assess the extent of receptor-ligand binding. Epilepsy models exhibited significant anxiety and depressive-like behaviors, and BA significantly reduced the severity of seizures in these rats while also alleviating their anxiety and depressive-like behaviors. Moreover, neuronal loss and damage were observed in the hippocampus of epileptic rats, but BA was able to effectively counteract this issue by enhancing BDNF expression and promoting neurogenesis within the hippocampus, especially in the DG region. The co-localization of Iba-1 with NLRP3 indicated the activation of NLRP3 inflammasome in microglia. Subsequent RT-PCR revealed that BA may alleviate anxiety and depressive-like behaviors in epileptic rats by activating the P2RX7/NLRP3/ IL-1β signaling pathway. The final molecular docking results indicated that BA had a good binding affinity with proteins, such as P2RX7, NLRP3, and IL-1β. This study confirmed the effectiveness of BA in improving anxiety and depressivelike behaviors associated with epilepsy. Moreover, it provides theoretical support for the neuroprotective role demonstrated by BA.

Mitochondria-targeting peptide SS-31 attenuates ferroptosis via inhibition of the p38 MAPK signaling pathway in the hippocampus of epileptic rats

Ferroptosis is a newly identified form of non-apoptotic regulated cell death (RCD) andplaysanimportantrole in epileptogenesis. The p38 mitogen-activated protein kinase (p38 MAPK) pathway has been confirmed to be involved in ferroptosis. The mitochondria-targeting antioxidant Elamipretide (SS-31) can reduce the generation of lipid peroxidation and the buildup of reactive oxygen species (ROS). Collectively, our present study was to decipher whether SS-31 inhibits ferroptosis via the p38 MAPK signaling pathway in the rat epilepsy model induced by pilocarpine (PILO).Adult male Wistar rats were randomly divided into four groups: control group (CON group), epilepsy group (EP group), SS-31 treatment group (SS group), and p38 MAPK inhibitor (SB203580) treatment group (SB group). Our results demonstrated that the rat hippocampal neurons after epilepsy were followed by accumulated iron and malondialdehyde (MDA) content, upregulated phosphorylated p38 MAPK protein (P-p38) and nuclear factor erythroid 2-related factor 2 (Nrf2) levels, reduced glutathione peroxidase 4 (Gpx4) content, and depleted glutathione (GSH) activity. Morphologically, mitochondrial ultrastructural damage under electron microscopy was manifested by a partial increase in outer membrane density, disappearance of mitochondrial cristae, and mitochondrial shrinkage. SS-31 and SB203580 treatment blocked the initiation and progression of ferroptosis in the hippocampus of epileptic rats via reducing the severity of epileptic seizures, reversing the expression of Gpx4, P-p38 , decreasing the levels of iron and MDA, as well as increasing the activity of GSH and Nrf2. To summarize, our findings proved that ferroptosis was coupled with the pathology of epilepsy, and SS-31 can inhibit PILO-induced seizures by preventing ferroptosis, which may be connected to the inhibition of p38 MAPK phosphorylation, highlighting the potential therapeutic value for targeting ferroptosis process in individuals with seizure-related diseases.

Role and mechanism of EphB3 in epileptic seizures and epileptogenesis through Kalirin

BackgroundThe EphB receptor tyrosine kinase family participates in intricate signaling pathways that orchestrate neural networks, guide neuronal axon development, and modulate synaptic plasticity through interactions with surface-bound ephrinB ligands. Additionally, Kalirin, a Rho guanine nucleotide exchange factor, is notably expressed in the postsynaptic membrane of excitatory neurons and plays a role in synaptic morphogenesis. This study postulates that Kalirin may act as a downstream effector of EphB3 in epilepsy. This investigation focuses on understanding the link between EphB3 and epilepsy. Materials and methodsChronic seizure models using LiCl-pilocarpine (LiCl/Pilo) and pentylenetetrazol were developed in rats. Neuronal excitability was gauged through whole-cell patch clamp recordings on rat hippocampal slices. Real-time PCR determined Kalirin's mRNA expression, and Western blotting was employed to quantify EphB3 and Kalirin protein levels. Moreover, dendritic spine density in epileptic rats was evaluated using Golgi staining. ResultsModulation of EphB3 functionality influenced acute seizure severity, latency duration, and frequency of spontaneous recurrent seizures. Golgi staining disclosed an EphB3-driven alteration in dendritic spine density within the hippocampus of epileptic rats, underscoring its pivotal role in the reconfiguration of hippocampal neural circuits. Furthermore, our data propose Kalirin as a prospective downstream mediator of the EphB3 receptor. ConclusionsOur findings elucidate that EphB3 impacts the action potential dynamics in isolated rat hippocampal slices and alters dendritic spine density in the inner molecular layer of epileptic rat hippocampi, likely through Kalirin-mediated pathways. This hints at EphB3's significant role in shaping excitatory circuit loops and recurrent seizure activity via Kalirin.

RelB mediated by GSK3β/IκBα as a potential therapeutic target in pilocarpine seizure model rats and drug-resistant epilepsy patients

BackgroundDrug-resistant epilepsy (DRE) affects more than 20 million people worldwide. DRE patients do not respond to anti-seizure medications. Shifting from anti-seizure to anti-epileptic and disease-modifying therapy will be an important aim for future research. The canonical nuclear factor kappa B (NF-κB) signaling pathway plays a pivotal role in epilepsy and neuroinflammation. However, the expression and regulation of RelB, which can be activated via both canonical and non-canonical NF-κB signaling, are obscure in epilepsy. To clarify the expression and localization of RelB in the DRE phenotype and to determine the proteins related to RelB regulation, we conducted the following studies. MethodsQuantitative PCR was performed to detect the transcription of RELB in pilocarpine-induced epileptic rats. Western blotting was used to determine the abundance of RelB and proteins related to RelB regulation in brain tissue from both epilepsy model rats and DRE patients and in liposaccharide-induced HT22 cells. Immunofluorescence staining and immunohistochemistry were used to locate RelB in brain sections from DRE patients. ELISA was used to determine inflammatory cytokines secreted by HT22 cells into the culture medium. ResultsRELB transcription and expression were increased in the hippocampus and cortex of epileptic rats during the acute and latent phases and in epileptic foci of patients with temporal lobe epilepsy. Additionally, RelB levels were mainly increased in epileptic rat neurons and accumulated in the nuclei of hippocampal neurons. Further research demonstrated increased GSK3β phosphorylation at the Ser9 inhibitory site and decreased IκBα levels, which contributed to RelB accumulation in hippocampal neurons after seizure and in liposaccharide-stimulated HT22 cells. ConclusionsThis study is the first to demonstrate that RelB is widely distributed and increased in epileptogenic foci neurons in epileptic phenotypes. These results indicate that RelB may play roles in DRE and is mediated by GSK3β and IκBα, providing a new target for anti-epileptic and disease-modifying therapy.

Assessment of Biochemical and Neuroactivities of Cultural Filtrate from Trichoderma harzianum in Adjusting Electrolytes and Neurotransmitters in Hippocampus of Epileptic Rats.

Epilepsy is a serious chronic neurological disorder, which is accompanied by recurrent seizures. Repeated seizures cause physical injuries and neuronal dysfunction and may be a risk of cancer and vascular diseases. However, many antiepileptic drugs (AEDs) have side effects of mood alteration or neurocognitive function, a reduction in neuron excitation, and the inhibition of normal activity. Therefore, the present study aimed to evaluate the effect of secondary metabolites of Trichoderma harzianum cultural filtrate (ThCF) when adjusting different electrolytes and neurotransmitters in the hippocampus of epileptic rats. Cytotoxicity of ThCF against LS-174T cancer cells was assessed using a sulforhodamine B (SRB) assay. Quantitative estimation for some neurotransmitters, electrolytes in sera or homogenate of hippocampi tissues, and mRNA gene expression for ion or voltage gates was assessed by quantitative Real-Time PCR. Treatment with ThCF reduces the proliferative percentage of LS-174T cells in a concentration-dependent manner. ThCF administration improves hyponatremia, hyperkalemia, and hypocalcemia in the sera of the epilepticus model. ThCF rebalances the elevated levels of many neurotransmitters and reduces the release of GABA and acetylcholine-esterase. Also, treatments with ThCF ameliorate the downregulation of mRNA gene expression for some gate receptors in hippocampal homogenate tissues and recorded a highly significant elevation in the expression of SCN1A, CACNA1S, and NMDA. Secondary metabolites of Trichoderma (ThCF) have cytotoxic activity against LS-174T (colorectal cancer cell line) and anxiolytic-like activity through a GABAergic mechanism of action and an increase in GABA as inhibitory amino acid in the selected brain regions and reduced levels of NMDA and DOPA. The present data suggested that ThCF may inhibit intracellular calcium accumulation by triggering the NAADP-mediated Ca2+ signaling pathway. Therefore, the present results suggested further studies on the molecular pathway for each metabolite of ThCF, e.g., 6-pentyl-α-pyrone (6-PP), harzianic acid (HA), and hydrophobin, as an alternative drug to mitigate the side effects of AEDs.

Inhibition of mTORC2 improves brain injury in epileptic rats by promoting chaperone-mediated autophagy

Epilepsy can seriously affect children's cognitive and behavioral development. The mechanistic target of rapamycin(mTOR) pathway plays an important role in neurodevelopment and epilepsy, but the mechanism of mechanistic target of rapamycin complex 2 (mTORC2) in epilepsy is still unclear. Here, we compared the similarities and differences of the mechanisms of action of mechanistic target of rapamycin complex 1 (mTORC1) and mTORC2 complex in the pathogenesis of epilepsy. Our research results show that the levels of apoptosis in cortical and hippocampal neurons were upregulated in epileptic rats (F = 32.15, 30.96; both P < 0.01), and epilepsy caused neuronal damage (F = 8.13, 9.43; both P < 0.01). The mTORC2-Akt pathway was activated in the cortex and hippocampus of epileptic rats. Inhibition of mTORC2 resulted in decreased levels of apoptosis and reduced neuronal damage in the cortex and hippocampus of epileptic rats. In the hippocampus, selective inhibition of mTORC2 increased lysosome-associated membrane protein 2 A (LAMP2A) protein expression compared with the control group, and the difference was statistically significant (F = 3.02, P < 0.05). Finally, we concluded that in the hippocampus, selective inhibition of mTORC2 can improve epileptic brain injury in rats by increasing chaperone-mediated autophagy (CMA) levels.

PtNPs/rGO-GluOx/mPD Directionally Electroplated Dual-Mode Microelectrode Arrays for Detecting the Synergistic Relationship between the Cortex and Hippocampus of Epileptic Rats.

Precise and directional couplings of functional nanomaterials with implantable microelectrode arrays (IMEAs) are critical for the manufacture of sensitive enzyme-based electrochemical neural sensors. However, there is a gap between the microscale of IMEA and conventional bioconjugation techniques for enzyme immobilization, which leads to a series of challenges such as limited sensitivity, signal crosstalk, and high detection voltage. Here, we developed a novel method using carboxylated graphene oxide (cGO) to directionally couple the glutamate oxidase (GluOx) biomolecules onto the neural microelectrode to monitor glutamate concentration and electrophysiology in the cortex and hippocampus of epileptic rats under RuBi-GABA modulation. The resulting glutamate IMEA exhibited good performance involving less signal crosstalk between microelectrodes, lower reaction potential (0.1 V), and higher linear sensitivity (141.00 ± 5.66 nA μM-1 mm-2). The excellent linearity ranged from 0.3 to 68 μM (R = 0.992), and the limit of detection was 0.3 μM. For epileptic rats, the proposed IMEA sensitively obtained synergetic variations in the action potential (Spike), local field potentials (LFPs), and glutamate of the cortex and hippocampus during seizure and RuBi-GABA inhibition. We found that the increase in glutamate preceded the burst of electrophysiological signals. At the same time, both changes in the hippocampus preceded the cortex. This reminded us that glutamate changes in the hippocampus could serve as important indicators for early warning of epilepsy. Our findings provided a new technical strategy for directionally stabilizing enzymes onto the IMEA with versatile implications for various biomolecules' modification and facilitated the development of detecting tools for understanding the neural mechanism.

Dehydroepiandrosterone Attenuates Astroglial Activation, Neuronal Loss and Dendritic Degeneration in Iron-Induced Post-Traumatic Epilepsy

Iron-induced experimental epilepsy in rodents reproduces features of post-traumatic epilepsy (PTE) in humans. The neural network of the brain seems to be highly affected during the course of epileptogenesis and determines the occurrence of sudden and recurrent seizures. The aim of the current study was to evaluate astroglial and neuronal response as well as dendritic arborization, and the spine density of pyramidal neurons in the cortex and hippocampus of epileptic rats. We also evaluated the effect of exogenous administration of a neuroactive steroid, dehydroepiandrosterone (DHEA), in epileptic rats. To induce epilepsy, male Wistar rats were given an intracortical injection of 100 mM solution (5 µL) of iron chloride (FeCl3). After 20 days, DHEA was administered intraperitoneally for 21 consecutive days. Results showed epileptic seizures and hippocampal Mossy Fibers (MFs) sprouting in epileptic rats, while DHEA treatment significantly reduced the MFs' sprouting. Astroglial activation and neuronal loss were subdued in rats that received DHEA compared to epileptic rats. Dendritic arborization and spine density of pyramidal neurons was diminished in epileptic rats, while DHEA treatment partially restored their normal morphology in the cortex and hippocampus regions of the brain. Overall, these findings suggest that DHEA's antiepileptic effects may contribute to alleviating astroglial activation and neuronal loss along with enhancing dendritic arborization and spine density in PTE.

Investigation of the effects of pinacidil and glibenclamide administration on HCN1, KCNT1, Kir 6.1, SUR1 gene expressions in hippocampus and cortex regions in epileptic rats

The purpose of this study was to look into the effects of pinacidil and glibenclamide on HCN1, KCNT1, Kir 6.1, and SUR1 gene expression in epileptic rats hippocampus and cortex. Male Wistar-Albino rats were used in this study. The drugs pinacidil and glibenclamide were utilized. Control, Epilepsy, Epilepsy-O, and Epilepsy-B were the five groups formed. The epileptic focus was created by intracortical administration of penicillin at a dose of 500 IU/2 μl. Hippocampus and Cortex are removed from all animals and Kir 6.1, SUR1, HCN1, and KCNT1 gene expression levels were determined by qPCR. The SPSS 21 program was used for statistics. HCN1 gene expression level is equal in the hippocampus and cortex (p&lt;0.05). KCNT1, SUR1, and KIR6.1 are more expressed in the hippocampus than in the cortex (p&lt;0.05). In epilepsy groups, HCN1 gene expression level was found to be higher than KCNT1, SUR1, and KIR6.1 gene expression levels (p&lt;0.05). KIR6.1, SUR1, gene expression levels decreased with the application of pinacidil and glibenclamide (p&lt;0.05). SUR1 and KIR6.1 gene expression levels were considerably lower in the epilepsy pinacidil group compared to the other groups. The gene expression levels in the hippocampus were found to be considerably higher than in the cortex group, according to this study. The fact that HCN1 gene expression levels are significantly greater in both the brain and the hippocampus 24 hours following the commencement of epileptic convulsions suggests that preventive medication may be possible.

Proteomic Analysis Reveals the Vital Role of Synaptic Plasticity in the Pathogenesis of Temporal Lobe Epilepsy.

Temporal lobe epilepsy (TLE) is a chronic neurological disorder that is often resistant to antiepileptic drugs. The pathogenesis of TLE is extremely complicated and remains elusive. Understanding the molecular mechanisms underlying TLE is crucial for its diagnosis and treatment. In the present study, a lithium-pilocarpine-induced TLE model was employed to reveal the pathological changes of hippocampus in rats. Hippocampal samples were taken for proteomic analysis at 2 weeks after the onset of spontaneous seizure (a chronic stage of epileptogenesis). Isobaric tag for relative and absolute quantization (iTRAQ) coupled with liquid chromatography-tandem mass spectrometry (LC–MS/MS) technique was applied for proteomic analysis of hippocampus. A total of 4173 proteins were identified from the hippocampi of epileptic rats and its control, of which 27 differentially expressed proteins (DEPs) were obtained with a fold change > 1.5 and P < 0.05. Bioinformatics analysis indicated 27 DEPs were mainly enriched in “regulation of synaptic plasticity and structure” and “calmodulin-dependent protein kinase activity,” which implicate synaptic remodeling may play a vital role in the pathogenesis of TLE. Consequently, the synaptic plasticity-related proteins and synaptic structure were investigated to verify it. It has been demonstrated that CaMKII-α, CaMKII-β, and GFAP were significant upregulated coincidently with proteomic analysis in the hippocampus of TLE rats. Moreover, the increased dendritic spines and hippocampal sclerosis further proved that synaptic plasticity involves in the development of TLE. The present study may help to understand the molecular mechanisms underlying epileptogenesis and provide a basis for further studies on synaptic plasticity in TLE.

Protective Effects of Intranasally Administrated Oxytocin-Loaded Nanoparticles on Pentylenetetrazole-Kindling Epilepsy in Terms of Seizure Severity, Memory, Neurogenesis, and Neuronal Damage.

Pentylenetetrazole (PTZ)-induced kindling is an animal model for studying human temporal lobe epilepsy (TLE), which is characterized by alterations of hippocampal neurons and memory. Although the intranasal (IN) administration of oxytocin (OT) has limited efficiency, nanoparticles (NPs) are a promising candidate to deliver OT to the brain. However, there are very limited data on epilepsy research about oxytocin-loaded nanoparticles (NP-OTs). The aim of this study is to investigate the effects of IN administration of chronic NP-OTs on the hippocampus of PTZ-induced male epileptic rats in terms of seizure severity, memory, neurogenesis, and neuronal damage. Saline/OT/NP-OTs were administrated to both control (Ctrl) and PTZ groups intranasally. Consequently, saline and PTZ were injected, respectively, 25 times every 48 h. Then, seizure severity (score and latency) was calculated for the PTZ groups. A spatial working memory evaluation test (SWMET) was performed after the last injection. Hippocampus histopathology, neurogenesis, and apoptosis were demonstrated. Serum total antioxidant status (TAS) and total oxidant status (TOS) levels and the oxidative stress index (OSI) were measured. We showed that OTs and NP-OTs prevented the kindling development and had positive effects on seizure severity. SWMET-related behaviors were also recovered in the PTZ + NP-OT group. A significant increase of neurogenesis and decrease of apoptosis in the hippocampus of the PTZ + NP-OT group were observed, while OTs and NP-OTs had protective effects against PTZ-induced damage to hippocampal neurons. Our results indicate that the chronic administration of NP-OTs may have positive effects on hippocampal damage via increasing neurogenesis and decreasing apoptosis and seizure severity.

Expression of matrix metalloproteinases and tissue inhibitors of metalloproteinases in the hippocampus of lithium-pilocarpine-induced acute epileptic rats.

Epilepsy is characterised by abnormal neuronal discharges, including aberrant expression of extracellular matrix (ECM) components and synaptic plasticity stabilisation. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) interact to remodel the ECM in the central nervous system (CNS), to modulate synaptic plasticity in epileptogenesis. In the present study, the expression of MMP activators (tPA and uPA), 10 MMPs, and 3 TIMPs was detected by western blot analysis and quantitative polymerase chain reaction (RT-qPCR) to assess their potential pathogenetic role in the epileptogenesis in the hippocampus of lithium-pilocarpine hydrochloride-induced epileptic rats. Our results showed that The expression of MMP7 and MMP14 was impeded in the hippocampus of lithium-pilocarpine-induced acute epileptic rats compared with that in controls. The transcriptional level of tPA was enhanced on day 1 post-seizure in the hippocampus, while the levels of several MMPs and TIMPs did not change on days 1 and 3 post-seizure compared with that in controls. The expression of MMPs and TIMPs reflects a novel feature of epileptogenesis and may offer new perspectives for future therapeutic interventions.

Alterações comportamentais e estruturais do hipocampo de ratos Wistar epilépticos são minimizadas pela acupuntura em associação ou não com fenobarbital

ABSTRACT The aim of this study was to analyze the behavior and histopathological changes in the hippocampus of epileptic Wistar rats treated with acupuncture associated or not with phenobarbital. The experiment used 44 male rats with 90 days of birth, induced to status epileptics with pilocarpine hydrochloride in a single dose of 350mg/kg, separated into treatment groups and submitted for 5 minutes to the elevated plus-maze test. Group 1 received 0.2mL of saline solution orally; Group 2 treated with acupuncture at the yintang, baihui, shishencong, jizhong, naohu, thianzu points; Group 3 received orally phenobarbital, daily dose of 20mg/kg; Group 4 treated with an association of acupuncture and oral phenobarbital; Group 5 random needling. The results obtained showed that Groups 2 (acupuncture) and 4 (acupuncture and phenobarbital) presented decreased anxiety, epileptic seizures, and neuronal death in the CA1, CA3 areas of the hippocampus when compared to animals in groups 1, 3 and 5. It is concluded that the association of phenobarbital and acupuncture points used in the experiment allowed for the control of epileptic seizures, reduction of anxiety and reduction of lesions in the subareas of the hippocampus.

Neuroprotective effects of 1,8-cineole on inhibition of apoptosis and Bcl-2/Caspase-3 mRNA expression in the hippocampus of Pilocarpine model epileptic rats

Neuroprotective effects of 1,8-cineole on inhibition of apoptosis and Bcl-2/Caspase-3 mRNA expression in the hippocampus of Pilocarpine model epileptic rats

Melissa officinalis L. ameliorates oxidative stress and inflammation and upregulates Nrf2/HO-1 signaling in the hippocampus of pilocarpine-induced rats.

Epilepsy is characterized by recurrent epileptic seizures, and its effective management continues to be a therapeutic challenge. Oxidative stress and local inflammatory response accompany the status epilepticus (SE). This study evaluated the effect of Melissa officinalis extract (MOE) on oxidative stress, inflammation, and neurotransmitters in the hippocampus of pilocarpine (PILO)-administered rats, pointing to the involvement of Nrf2/HO-1 signaling. Rats received PILO via intraperitoneal administration and were treated with MOE for 2 weeks. MOE prevented neuronal loss; decreased lipid peroxidation, Cox-2, PGE2, and BDNF; and downregulated glial fibrillary acidic protein in the hippocampus of PILO-treated rats. In addition, MOE enhanced GSH and antioxidant enzymes, upregulated Nrf2 and HO-1 mRNA abundance, and increased the nuclear translocation of Nrf2 in the hippocampus of epileptic rats. Na+/K+-ATPase activity and GABA were increased, and glutamate and acetylcholine were decreased in the hippocampus of epileptic rats treated with MOE. In conclusion, MOE attenuated neuronal loss, oxidative stress, and inflammation; activated Nrf2/HO-1 signaling; and modulated neurotransmitters, GFAP, and Na+/K+-ATPase in the hippocampus of epileptic rats. These findings suggest that M. officinalis can mitigateepileptogenesis, pending further studies to explore the exact underlying mechanisms.

Low-Intensity Focused Ultrasound-Mediated Attenuation of Acute Seizure Activity Based on EEG Brain Functional Connectivity.

(1) Background: Ultrasound has been used for noninvasive stimulation and is a promising technique for treating neurological diseases. Epilepsy is a common neurological disorder, that is attributed to uncontrollable abnormal neuronal hyperexcitability. Abnormal synchronized activities can be observed across multiple brain regions during a seizure. (2) Methods: we used low-intensity focused ultrasound (LIFU) to sonicate the brains of epileptic rats, analyzed the EEG functional brain network to explore the effect of LIFU on the epileptic brain network, and continued to explore the mechanism of ultrasound neuromodulation. LIFU was used in the hippocampus of epileptic rats in which a seizure was induced by kainic acid. (3) Results: By comparing the brain network characteristics before and after sonication, we found that LIFU significantly impacted the functional brain network, especially in the low-frequency band. The brain network connection strength across multiple brain regions significantly decreased after sonication compared to the connection strength in the control group. The brain network indicators (the path length, clustering coefficient, small-worldness, local efficiency and global efficiency) all changed significantly in the low-frequency. (4) Conclusions: These results revealed that LIFU could reduce the network connections of epilepsy circuits and change the structure of the brain network at the whole-brain level.

Increased immunoreactivity of glutamate receptors, neuronal nuclear protein and glial fibrillary acidic protein in the hippocampus of epileptic rats with fast ripple activity.

Epilepsy is a neurological disorder in which an imbalance between excitatory and inhibitory transmission is observed. Glutamate is the principal excitatory neurotransmitter that acts through ionic and metabotropic receptors; both types of receptors are involved in temporal lobe epilepsy (TLE). High frequency oscillations called fast ripples (FR, 250-600Hz) have been observed, particularly in the hippocampus, and they are involved in epileptogenesis. The present study analyzed the immunoreactivity of the principal glutamate receptors associated with epilepsy in epileptic animals with FR activity. Male Swiss-Wistar rats (210-250 gr) were injected with pilocarpine (2.4mg/2µl) and were video monitored (24/7) until the appearance of spontaneous and recurrent seizures. Then, a deep microelectrode implantation surgery was performed in the DG, CA3 and CA1 regions, and FR activity was observed 1-, 2-, 3-, 7-, and 14-day postsurgery. The animals were sacrificed on day 15, and fluorescence immunohistochemistry was carried out in the hippocampus for the α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), N-methyl-D-aspartate (NMDA) and mGlu-R5 glutamate receptors as well as Neuronal Nuclear Protein (NeuN) and Glial Fibrillary Acidic Protein (GFAP). An increase in the immunoreactivity for the three receptors was found. However, the AMPA receptor showed an increase in the three regions analyzed (i.e., DG, CA1 and CA3). The findings showed a decrease of NeuN in the DG and an increase of GFAP. These results suggest an important role of glutamate receptors in the hippocampus of epileptic rats with FR activity.

Mechanisms of microRNA‑142 in mitochondrial autophagy and hippocampal damage in a rat model of epilepsy.

Researchers have confirmed the microRNA (miRNA/miR)-epilepsy association in rodent models of human epilepsy via a comprehensive database. However, the mechanisms of miR-142 in epilepsy have not been extensively studied. In the present study, a rat model of epilepsy was first established by an injection of lithium chloride-pilocarpine and the successful establishment of the model was verified via electroencephalogram monitoring. The levels of miR-142, phosphatase and tensin homolog deleted on chromosome 10 (PTEN)-induced putative kinase 1 (PINK1), marker proteins of mitochondrial autophagy, and apoptosis-related proteins were measured. Additionally, the pathological changes in the hippocampus, the ultrastructure of the mitochondria, and degeneration and the apoptosis of neurons were observed using different staining methods. The malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the hippocampus, mitochondrial membrane potential (MTP) and reactive oxygen species (ROS) generation were detected. Furthermore, the targeting association between miR-142 and PINK1 was predicted and verified. Consequently, apoptosis increased, and mitochondrial autophagy decreased, in the hippocampus of epileptic rats. Following miR-142 inhibition, the epileptic rats exhibited an increased Bax expression, a decreased Bcl-2 expression, upregulated marker protein levels of mitochondrial autophagy, a reduced MDA content, an enhanced SOD activity, an increased MTP and decreased ROS generation. PINK1 is a target gene of miR-142, and its overexpression protected against hippocampal damage. Taken together, the results of the present study demonstrated that miR-142 inhibition promotes mitochondrial autophagy and reduces hippocampal damage in epileptic rats by targeting PINK1. These findings may provide useful information for the treatment of epilepsy.

Short-Term Amygdala Low-Frequency Stimulation Does not Influence Hippocampal Interneuron Changes Observed in the Pilocarpine Model of Epilepsy.

Temporal lobe epilepsy (TLE) is characterized by changes in interneuron numbers in the hippocampus. Deep brain stimulation (DBS) is an emerging tool to treat TLE seizures, although its mechanisms are not fully deciphered. We aimed to depict the effect of amygdala DBS on the density of the most common interneuron types in the CA1 hippocampal subfield in the lithium-pilocarpine model of epilepsy. Status epilepticus was induced in male Wistar rats. Eight weeks later, a stimulation electrode was implanted to the left basolateral amygdala of both pilocarpine-treated (Pilo, n = 14) and age-matched control rats (n = 12). Ten Pilo and 4 control animals received for 10 days 4 daily packages of 50 s 4 Hz regular stimulation trains. At the end of the stimulation period, interneurons were identified by immunolabeling for parvalbumin (PV), neuropeptide Y (NPY), and neuronal nitric oxide synthase (nNOS). Cell density was determined in the CA1 subfield of the hippocampus using confocal microscopy. We found that PV+ cell density was preserved in pilocarpine-treated rats, while the NPY+/nNOS+ cell density decreased significantly. The amygdala DBS did not significantly change the cell density in healthy or in epileptic animals. We conclude that DBS with low frequency applied for 10 days does not influence interneuron cell density changes in the hippocampus of epileptic rats.

The Expression of ZnT3 and GFAP Is Potentiated in the Hippocampus of Drug-Resistant Epileptic Rats Induced by Amygdala Kindling

Objective: The first-line treatment for epilepsy, a chronic neurological disorder characterized by spontaneous seizures, includes the application of anticonvulsant drug therapy. Only one-third of patients are incapable of complete controlling of their seizures after the administration of ≥2 pharmaceuticals. Here, we aimed to observe the ultrastructure changes and the expression of ZnT3 and GFAP in the hippocampus of drug-resistant epileptic rats. Methods: A total of 50 healthy adult male SD rats were used to generate the model ofepilepsy by amygdala kindling. After the rats were successfully kindled, pharmacoresistant epileptic (PRE) rats were selected according to their response to phenobarbital and phenytoin. The ultrastructure as well as the expression of zinc transporter 3 (ZnT3, a member of a growing family of mammalian zinc transporters) and glial fibrillary acidic protein (GFAP) were compared among PRE, pharmacosensitive epileptic (PRE), and normal (NRC) rats. Results: The PRE rats displayed severe synapses, neuronal degeneration, and necrosis. Moreover, the expression of ZnT3 and GFAP was significantly increased in both PRE and PSE rats; compared with NRC rats, the promotion of this expression was more pronounced in the PRE rats. Conclusions: Taken together, obvious synapses, neuronal degeneration, necrosis, mossy fiber sprouting, and astrogliosis were found in the drug-resistant epileptic rat model induced by amygdala kindling.