Kindling Model Research Articles

Reduction of seizures and inflammatory markers by betamethasone in a kindling seizure model.

Epilepsy is a chronic disease characterized by an ongoing predisposition to seizures. Although inflammation has emerged as a crucial factor in the etiology of epilepsy, no approaches to anti-inflammatory treatment have been clinically proven to date. Betamethasone (a corticosteroid drug used in the clinic for its anti-inflammatory and immunosuppressive effects) has never been evaluated in attenuating the intensity of seizures in a kindling animal model of seizures. Using a kindling model in male wistar rats, this study evaluated the effect of betamethasone on the severity of seizures and levels of pro-inflammatory interleukins. Seizures were induced by pentylenetetrazole (30mg/kg) on alternate days for 15days. The animals were divided into four groups: a control group treated with saline, another control group treated with diazepam (2mg/kg), and two groups treated with betamethasone (0.125 and 0.250mg/kg, respectively). Open field test was conducted. Betamethasone treatments were effective in reducing the intensity of epileptic seizures. There were lower levels of Tumor Necrosis Factor-α and interleukin-1β in the cortex, compared to the saline group, on the other hand, levels in the hippocampus remained similar to the control groups. There was no change in the levels of interleukin-6 in the evaluated structures. Serum inflammatory mediators remained similar. Lower quantities of inflammatory mediators in the central nervous system may have been the key to the reduced severity of seizures on the Racine scale.

Dynamic changes of striatal neurotransmitter systems activity cause changes in behavioral non-convulsive disorders of depressive nature in the course of chronic seizure activity formation in kindling model of epileptogenesis

Epilepsy is a chronic neurological disease characterized by recurrent behavioral seizures and affects approximately 1% of the global population. Epilepsy causes recurrent behavioral seizures, which are temporary behavioral changes caused by disordered, synchronized, and rhythmic activations of specific populations of neurons in the brain. Depressive disorders are one of the most common comorbid behavioral disorders in patients with epilepsy, yet they remain undiagnosed and untreated. The aim of the study is to investigate the severity of non-convulsive swimming and emotional behavior and mnemonic processes in rats with a kindling model of epilepsy under conditions of modulation of neurotransmitter systems of the caudate nuclei. It was found that in the dynamics of picrotoxin-induced kindling in rats, impaired swimming behavior, learning and memory abilities, as well as emotional dysfunctions, which are correlates of non-convulsive behavior, are recorded. It has been proven that the detected disorders of non-convulsive behaviors progressed in the dynamics of the formation of the kindling model of epilepsy and were maximal at the stages of completed kindling and postkindling. The obtained data demonstrate the key role of the striatum in the formation of non-convulsive behavioral disorders. On the 18th day, the pro-convulsive role of striatum is noted, which is confirmed by the enhancement of GABAergic and inhibition of dopaminergic neurotransmission of striatum along with the inhibition of its cholinergic mediation. The completion of the formation of kindling is confirmed by the enhancement of cholinergic and dopaminergic neurotransmission of striatum along with the inhibition of GABAergic mechanisms. In the postkindling stage, activation of cholinergic and dopaminergic neurotransmission of striatum and inhibition of its GABAergic activity are observed. The author considers the data obtained to be an experimental basis for the feasibility of testing clinical diagnostic effects in terms of dynamic monitoring of the behavior of patients with epilepsy during interictal periods, as well as possible corrective pharmacological effects in the case of modulation of the activity of certain neurotransmitter systems and intrastriatal neurotransmission.

Tetramethylpyrazine contributes to the neuroprotection in a rodent epileptic model of pentylenetetrazole-induced kindling.

In this study, tetramethylpyrazine (TMP) was evaluated for its therapeutic potential as an alternative therapy for epileptogenesis and its associated comorbidities in rats. The sub-convulsant dose of pentylenetetrazole (PTZ) (35 mg/kg, intraperitoneally) was injected on alternative days to produce kindling for 32 days and observed for seizure score percent of kindled animals in each group. After kindling, the animals were evaluated in models of anxiety, memory and predictive of depression. The neuroprotective effect of TMP was assessed by estimating the biochemical parameters in the cortex and hippocampus of the brain. Histopathological alterations were also observed in the cortex and hippocampus (CA1, CA3 and DG). The administration of TMP reduced the seizure score and percentage of kindled animals dose-dependently. Furthermore, TMP significantly improved the behavioural parameters measured in the predictive models of depression but not in the anxiety and cognitive performances of the animals. The oxidative-nitrosative stress, excitotoxicity, neuroinflammation and histological alterations in the brain induced by PTZ were significantly mitigated by administering the TMP high dose of 60 mg/kg. In conclusion, the TMP attenuated the depression behaviour in the PTZ-induced kindled rats, and reduced the oxidative-nitrosative stress, excitotoxicity, neuroinflammation and histological alterations of the brain.

Neurodegenrative changes in retina of rats with chronic epileptic syndrome under conditions of treatment with niacin-oxy-ethylidene-phosphonate germanate (MIGU-4)

Background:Epilepsy is accompanied by neurodegenerative changes, particularly those in the retina. Elucidation of the mechanisms of retinal alterations in a model of epileptic syndrome may allow for the development of new approaches to pharmacological vision correction. Purpose: To assess the morphological characteristics of the retina, particularly treated with niacin oxy-ethylidene phosphonate germinate (MIGU-4), in the pentylenetetrazole (PTZ)-induced kindling model of chronic epileptic syndrome. Material and Methods: PTZ was administered in Wistar male rats intraperitoneally (i.p.) at a daily dose of 35.0 mg/kg for 21 days. Rats with fully-developed generalized seizures were treated with MIGU-4 ip at a daily dose of 5.0 mg/kg or 25.0 mg/kg for 28 days. Thereafter, the animals were euthanized, their globes were enucleated, and retinal sections were prepared and stained with hematoxylin and eosin for morphological examination. Results: Cell density in the retinal ganglion cell layer was 2.14 times lower, and in the inner nuclear layer and outer nuclear layer, 41.0% and 19.0%, respectively, lower for the rats with fully developed kindled seizures than for controls (р less 0.05) In the presence of treatment with MIGU-4 ip at a daily dose of 25.0 mg/kg, cell density in the retinal ganglion cell layer and in the inner nuclear layer was 38% and 30,5%, respectively, higher than for controls (р less 0,05). There was no significant difference in the cell density in the outer nuclear layer between rats treated with MIGU-4 ip at a daily dose of 5 mg/kg or 25,0 mg/kg and controls (р more 0.05). Conclusion: The PTZ-induced kindling model of chronic epileptic syndrome is accompanied by degenerative changes in the eye. A course of treatment with MIGU-4 causes neuroptotective effects in a model of PTZ-induced retinopathy.

TC-G 1008 facilitates epileptogenesis by acting selectively at the GPR39 receptor but non-selectively activates CREB in the hippocampus of pentylenetetrazole-kindled mice

The pharmacological activation of the GPR39 receptor has been proposed as a novel strategy for treating seizures; however, this hypothesis has not been verified experimentally. TC-G 1008 is a small molecule agonist increasingly used to study GPR39 receptor function but has not been validated using gene knockout. Our aim was to assess whether TC-G 1008 produces anti-seizure/anti-epileptogenic effects in vivo and whether the effects are mediated by GPR39. To obtain this goal we utilized various animal models of seizures/epileptogenesis and GPR39 knockout mice model. Generally, TC-G 1008 exacerbated behavioral seizures. Furthermore, it increased the mean duration of local field potential recordings in response to pentylenetetrazole (PTZ) in zebrafish larvae. It facilitated the development of epileptogenesis in the PTZ-induced kindling model of epilepsy in mice. We demonstrated that TC-G 1008 aggravated PTZ-epileptogenesis by selectively acting at GPR39. However, a concomitant analysis of the downstream effects on the cyclic-AMP-response element binding protein in the hippocampus of GPR39 knockout mice suggested that the molecule also acts via other targets. Our data argue against GPR39 activation being a viable therapeutic strategy for treating epilepsy and suggest investigating whether TC-G 1008 is a selective agonist of the GPR39 receptor.

Tauroursodeoxycholic Acid (TUDCA) Regulates Inflammation and Hypoxia in Autonomic Tissues of Rats with Seizures.

Inflammation and hypoxia have an effect on the molecular mechanism of cardiovascular and respiratory pathologies accompanying seizures. Against this, Tauroursodeoxycholic Acid (TUDCA) can regulate oxidative stress, inflammation and cellular survival by suppressing endoplasmic reticulum (ER) stress. We evaluated the expression changes of NF-κB p65, TNF-α, HIF1α and Kir6.2 proteins associated with seizures in brain stem, heart and lung tissues representing the autonomous network. Additionally, we examined the protective effects of TUDCA administration against damage caused by seizures in terms of immunohistochemistry and pathology. 4 groups of Wistar Albino male rats (250-300 g, n=32) were formed as control, pentylenetetrazole (PTZ), TUDCA and PTZ+TUDCA. The epilepsy kindling model was created by intraperitoneal (i.p.) injection of PTZ chemical (35 mg/kg, every 2 days) for one month. TUDCA (500 mg/kg; every 2 days) treatment was given intraperitoneally 30 minutes before seizures for 1 month. Brain stem, heart (atria, ventricle) and lung tissues of rats were isolated. NF-κB p65, TNF-α, HIF1α and Kir6.2 proteins in the obtained tissues were evaluated by immunohistochemical staining. The immunoreactivity of the investigated proteins in the brainstem heart and lung tissues of rats with chronic PTZ administration was significantly increased. Recurrent seizures led to accumulation of inflammatory cells in tissues, hemorrhage, vasodilation, and apoptosis. Following TUDCA administration, expression of NF-κB p65, TNF-α and Kir6.2 was significantly reduced in all tissues (except the atrium of the heart) compared to control rats. HIF-1α levels were significantly suppressed in ventricular and lung tissues of epileptic rats given TUDCA. However, TUDCA pretreatment improved histopathological changes due to chronic seizures and partially reduced apoptosis. We showed that epileptic seizures may cause tissue damage with the development of inflammatory and hypoxic conditions in the brainstem and organs that represent the autonomic network. TUDCA therapy could be an effective agent in the treatment of cardiac and respiratory problems associated with seizures.

CRISPR-Based KCC2 Upregulation Attenuates Drug-Resistant Seizure in Mouse Models of Epilepsy.

The precise intervention of K-Cl cotransporter isoform 2 (KCC2) as a promising target for drug-resistant epilepsy remains elusive. Here, we used a CRISPRa system delivered by adeno-associated viruses to specifically upregulate KCC2 in the subiculum to confirm its therapeutic potential in various in vivo epilepsy models. Calcium fiber photometry was used to reveal the role of KCC2 in the restoration of impaired GABAergic inhibition. CRISPRa system effectively upregulated KCC2 expression both in in vitro cell culture and in vivo brain region. Delivery of CRISPRa with adeno-associated viruses resulted in upregulating the subicular KCC2 level, contributing to alleviating the severity of hippocampal seizure and facilitating the anti-seizure effect of diazepam in a hippocampal kindling model. In a kainic acid-induced epilepticus status model, KCC2 upregulation greatly increased the termination percentage of diazepam-resistant epilepticus status with the broadened therapeutic window. More importantly, KCC2 upregulation attenuated valproate-resistant spontaneous seizure in a kainic acid-induced chronic epilepsy model. Finally, calcium fiber photometry showed CRISPRa-mediated KCC2 upregulation partially restored the impaired GABAA -mediated inhibition in epilepsy. These results showed the translational potential of adeno-associated viruses-mediated delivery of CRISPRa for treating neurological disorders by modulating abnormal gene expression that is directly associated with neuronal excitability, validating KCC2 as a promising therapeutic target for treating drug-resistant epilepsy. ANN NEUROL 2023.

Chemical composition and anticonvulsant activities of herb pair of Gastrodia elata Blume-Acorus tatarinowii Schott decoction on experimentally induced seizures in mice.

Epilepsy is a serious public health problem in the world. At present, over 30% of affected patients remain refractory to currently available treatment. Medicinal plants as pharmaceuticals and healthcare treatments have been frequently used in the management of epilepsy in China for many centuries. Gastrodia elata-Acous tatarinowii (GEAT), as a classic and most commonly used herb pair in traditional Chinese medicine (TCM), has been employed to control seizures for thousands of years. However, the animal experiment data on its anticonvulsant effect is limited in the literature. Thus, this study aimed to reveal the therapeutic actions of GEAT decoction against seizures in mice. UHPLC-MS/MS was performed to analyze the chemical components of GEAT decoction. The mice were given GEAT decoction for 7days, and MES, PTZ, and 3-MP injection was given 30min after the last administration. Video monitoring was performed for comparisons. In addition, the PTZ-induced kindling models were conducted to investigate the seizure severity, anxiety and cognitive profile, inflammation, and oxidative stress parameters in mice. The results showed that GEAT decoction dose-dependently protected mice against MES, 3-MP, and PTZ-induced acute seizures. Furthermore, GEAT decoction significantly ameliorated seizure severity, decreased the accumulation of inflammatory mediators TNF-α, IL-1β, and IL-6, mitigated oxidative stress, as well as alleviated anxious-like behavior and cognitive deficits in PTZ-kindled mice. These results suggest that GEAT decoction possesses certain anticonvulsant properties, which might be clinically useful as phytotherapy alone or as an adjunct therapy for the prevention and treatment of seizures and epilepsy.

Bibliometric Analysis of the Published Studies on the Kindling Model between 1980 and 2023

Objective: Kindling is an animal model of epilepsy induced by electrical stimulation of the brain. The present study aimed to present a different perspective with a bibliometric approach by using the literature data on the “Kindling model” related keywords in the Web of Science (WoS) online database between 1980 and 2023.
 Methods: The bibliometric data were obtained from the online database WoS and analyzed and visualized with the VoS Viewer Program. The bibliometric datasets were analyzed and visualized regarding article productivity numbers according to years, article productivity numbers according to countries, the most used keywords according to authors, and cross-country cooperation.
 Result: Considering the results of the analysis of the published datasets, 2022 was determined as the year with the highest article productivity, and an acceleration was observed in the publication increase rate on the subject in general. When the order of the countries in the top three in the number of article productivity was examined, the USA, Germany, and Japan are the main countries, respectively. The most used keywords by the authors were determined as “Epilepsy”, “Kindling”, and “Hippocampus”. In the cooperation among countries, it was found that the USA, Germany, and Japan had more cooperation with other countries, respectively.
 Conclusıon: This study will contribute to the literature by providing a detailed understanding of the research basis, relevant research results, current research boundaries and main research focus in the Kindling Model.

Syntaxin 7 modulates seizure activity in epilepsy

The exact pathogenesis of epilepsy, one of the most common and devastating diseases of the nervous system, is not fully understood. Syntaxin7 (STX7) is a member of the SNARE superfamily, which mediates membrane fusion events in all cells. However, the role STX7 plays in epilepsy remains unclear. Therefore, this study investigates the role of STX7 in epilepsy. Our study found that the expression of STX7 was reduced in the epileptic brain and that overexpression of STX7 decreased the susceptibility to epileptic seizures and alleviated epileptic activity in a kainic acid–induced model and pentylenetetrazole-induced kindling model of epilepsy, whereas the downregulation of STX7 showed opposite effects. Whole-cell patch-clamp recordings showed that STX7 does not affect the intrinsic excitability of neurons, but rather the excitation/inhibition ratio mediated by affecting the release of presynaptic γ-aminobutyric acid neurotransmitters. Transmission electron microscopy results showed that STX7 did not affect the density of inhibitory synapses but could affect the density of inhibitory vesicles. Taken together, these results reveal a previously unknown function of STX7 in epilepsy and suggest that STX7 may serve as a novel target for epilepsy therapy.

Temporal stability of neurophysiological drug cue reactivity before and after acute stress in cannabis users: A test of incentive sensitization

BackgroundGiven increasing rates of Cannabis Use Disorder (CUD), objective measures are needed that can reliably index risk and track cannabis use progression. Based on incentive sensitization models, neurophysiological reactivity to cannabis cues, measured with the electroencephalography-recorded late positive potential (LPP), may be a candidate biomarker. To serve as such, the cannabis cue-elicited LPP must demonstrate adequate retest reliability and sensitivity to cannabis use change. Moreover, incentive sensitization theory suggests that state-level contextual variables, such as acute stress, can impact drug cue reactivity. Therefore, the present study evaluated the three-month retest reliability of the cannabis cue-elicited LPP, recorded before and after a laboratory stress induction, as well as its sensitivity to cannabis use change. MethodCannabis and neutral cue-elicited LPPs were measured in 102 adults reporting frequent cannabis use (86 % with current CUD) before and after an acute stress induction at two lab visits three-months apart. Physiological and subjective stress reactivity were also measured. ResultsManipulation checks confirmed expected cannabis cue and acute stress effects. Cannabis cue-elicited LPP amplitudes showed significant three-month retest reliability of poor-to-fair through moderate-to-good size. Change in cannabis use frequency significantly predicted change in cannabis cue-elicited LPP amplitudes, particularly at post-stress. ConclusionConsistent with incentive sensitization models of addiction, the cannabis cue-elicited LPP demonstrated trait-like, moderate three-month stability and responsivity to change in cannabis use behavior. Greater predictive validity of the post-stress LPP may arise through kindling effects of acute stress on incentive salience-related neural activity, which should be explored in future studies.

Assessment of in vivo antiepileptic potential and phytochemical analysis of Cassia absus seed extracts

Cassia absus, a member of Fabaceae family, has been a part of traditional medicine for various ailments such as Hypertension, Diabetes, and Cancer. This family of plants has been utilized for Anticonvulsant and Anxiolytic effects. The ongoing investigation is aimed to seek the antiepileptic potential of C. absus seed extracts in pentylenetetrazole-induced kindling mice. The seeds of C. absus were subjected to a sequential extraction process for the preparation of n-hexane, chloroform, methanol, and aqueous extracts. The PTZ-induced kindling model was employed to assess the antiepileptic activity of each extract. Seizure activity and antioxidant biomarkers in the brain tissue such as levels of CAT, SOD, tGSH, and MDA were assessed. Mechanism of action was elucidated by Flumazenil. Through GC-MS analysis, the phytochemical components in the chloroform extract of C. absus were evaluated.The outcomes showed that C. absus extracts markedly reduced the seizure activity in kindling mice. The extracts exhibited significant Antioxidant properties by enhancing the levels of antioxidant biomarkers in the brain tissue such as CAT, SOD, and tGSH, and decreasing the MDA level. The results demonstrated that C. absus extracts showed antiepileptic effects may be via GABA pathway.According to the results of this investigation, C. absus has significant antiepileptic potential in PTZ-induced kindling mice via GABA pathway modulation and combating reactive oxygen species.

The effects of kindling during pregnancy on long-term potentiation (LTP) induction and M1 muscarinic acetylcholine receptors in male rat offspring.

Neonates of pregnant women with epilepsy may compromise normal neurodevelopment and hippocampal morphology. Memory and learning disorders and a decrease in verbal IQ scores are seen in these children later in life. In the previous study, we suggested that the central muscarinic cholinergic receptors had an important role in learning and memory deficits induced by prenatal pentylenetetrazol-kindling in pups born to kindled mothers. This study aimed to investigate the effects of kindling during pregnancy on long-term potentiation (LTP) induction and the role of M1 muscarinic acetylcholine receptors in the hippocampus of male offspring. Twenty female Wistar rats were divided into two groups on the 13th day of their gestation (kindled and control; n = 10). Animals in the first group were kindled by i.p. injections of 25 mg/kg body weight pentylenetetrazol every 15 min until seizures occurred and the control group received normal saline. The effect of maternal seizures and perfusion of specific M1 muscarinic receptors antagonist (telenzepine at doses of 0.01, 0.1, and 1 nmol) on the LTP induction of 80 pups were tested at 12 weeks of age by field potential recordings. The results of the electrophysiological study revealed that recurrent seizures during pregnancy impaired field excitatory postsynaptic potentials (fEPSP)-LTP induction and normal development of M1 muscarinic receptors in the hippocampus of male offspring. Also, the results demonstrated that maternal seizure did not significantly affect the paired-pulse indexes and population spike-LTP in the hippocampus of male offspring. Our study showed that recurrent seizures during pregnancy cause impaired fEPSP-LTP induction and abnormal development of the M1 muscarinic receptor in the hippocampus.

Modified Atkins Diet Delayed the Onset of Epileptogenesis, Improved Motor Coordination and Enhanced Learning Memory in Mice

Modified Atkins diet is a high fat, moderate protein and carbohydrate diet that has long been used to manage intractable epilepsy in developed countries. However, the use of this diet for same condition is uncommon in most low- and middle-income countries including Nigeria. In a previous research, we formulated modified Atkins diet using cheap and locally-available food materials in Nigeria. This research is thus, aimed at evaluating the effect of the formulated modified Atkins diet on seizure plasticity using the PTZ-induced kindling model of seizure. We also investigated the effect of sub-chronic use (60 days) of the diet on motor coordination using positional sense test and on visual-spatial reference and learning memory using Morris-water maze. The result revealed that the formulated modified Atkins diet significantly (<i>p<0.05</i>) increased seizure plasticity by reducing the severity of seizure as well as delaying the onset of tonic-clonic seizure and death. The diet also enhanced motor coordination, and improved learning memory, but not reference memory in mice. We therefore concluded that the formulated modified Atkins diet may be of benefit in the management of intractable/treatment-resistant epilepsy, especially in patients who are already suffering from amnesia and/or postural instability since the diet was able to offer protection against seizure-induced memory loss and motor-incoordination in mice.

Life stress and Bipolar Disorder: regarding a clinical case

IntroductionResearch on life stress in bipolar disorder largely fails to account for the possibility of a dynamic relationship between psychosocial stress and episode initiation. The kindling hypothesis states that over the course of recurrent affective disorders, there is a weakening temporal relationship between major life stress and episode initiation that could reflect either a progressive sensitization or progressive autonomy to life stress.ObjectivesTo explore the concept of the Kindling model applied to bipolar disorder and to present a clinical case of a bipolar patient whose latter mood episodes were caused by adverse life events.MethodsWe performed a non-systematic literature review using the most relevant papers found on the database PubMed with the keywords “kindling effect”, “allostatic load”, “bipolar disorder” and “prevention”. Description of the clinical case report.ResultsThe phenomenon of kindling was first discovered by Goddard in 1967 who described it in epilepsy. Later, Post applied it to the bipolar disorder, arguing that the initial episodes of both unipolar and bipolar affective disorders are often precipitated by psychosocial stressors, but after multiple recurrences, not only do precipitated episodes continue to occur, but so do spontaneous ones as well. We present the case report of a 62 years old woman, divorced, diagnosed with type 1 bipolar disorder since she was 20 years old. She always have had poor adherence to her medication and follow-up with Psychiatry consultation, with a non-containing sociofamily environment that does not promote clinical stability. Over the time, her admissions on the Psychiatry ward were more frequent and precipitated by adverse life events, mainly caused by the deteriorated relationship with her children.ConclusionsThe kindling model clarifies aspects of the longitudinal course of episode development, recurrence, and progression to spontaneity, as well as further conceptual and theoretical rationales for intervention in order to prevent illness progression.Disclosure of InterestNone Declared

Do prospective longitudinal studies of bipolar disorder support the hypothesis of neuroprogression?

IntroductionBipolar I disorder is a mental disorder with the risk of severe clinical outcomes. Bipolar disorder was initially defined based on having a better outcome than schizophrenia. However, while recent longer-term findings in schizophrenia do not support neuroprogression, bipolar disorder is increasingly depicted as having neuroprogressive elements. There are, however, remarkably few prospective longitudinal studies of representative bipolar I cohorts followed from the first treatment.ObjectivesTo study the clinical development of a representative cohort of bipolar disorder patients recruited at their first treatment.MethodsPatients with DSM-IV Bipolar I or Bipolar NOS were consecutively recruited from in-and outpatient units in the larger Oslo area during their first treatment year and extensively clinically characterized at baseline. They then participated in personal one- and ten-year follow-ups.ResultsSixty-nine patients participated in the 10-year follow-up. Age at follow-up was 39.0 (+ 9.6) years, 59% were females. A total of 12% had unipolar mania, 58% had psychotic bipolar disorder, and 20% had experienced rapid cycling. At follow-up, 75% were in full affective remission, 60% had regained full functioning, and 54% were in stable full recovery.Mood episode relapses clustered around the first episode. Despite occasional relapses, 2/3 were mainly euthymic during the follow-up period. A small sub-group was highly affected from the first 2-3 years of treatment, but there were no apparent signs of kindling effects or indications of neuroprogressionConclusionsThe follow-up of this cohort of first-treatment Bipolar I patients does not support the hypothesis of neuroprogression.Disclosure of InterestNone Declared

Neuroprotection impact of biochanin A against pentylenetetrazol-kindled mice: Targeting NLRP3 inflammasome/TXNIP pathway and autophagy modulation.

Recurrent seizures characterize epilepsy, a complicated and multifaceted neurological disease. Several neurological alterations, such as cell death and the growth of gorse fibers, have been linked to epilepsy. The dentate gyrus of the hippocampus is particularly vulnerable to neuronal loss and abnormal neuroplastic changes in the pentylenetetrazol (PTZ) kindling model. Biochanin A has potent anti-inflammatory and antioxidant properties, according to previous evidence and its possible impact in epilepsy has never previously been claimed. The current work aimed to investigate biochanin A's anti-epileptic potential in PTZ-induced kindling model in mice. Chronic epilepsy was established in mice by giving PTZ (35mg/kg, i.p) every other day for 21days. Biochanin A (20mg/kg) was given daily till the end of the experiment. Biochanin A pretreatment significantly reduced the severity of epileptogenesis by 51.7% and downregulated the histological changes in the CA3 region of the hippocampus by 42% along with displaying antioxidant/anti-inflammatory efficacy through upregulated hemeoxygenase-1 (HO-1) and, erythroid 2-related factor 2 (Nrf2) levels in the brain by 1.9-fold and 2-fold respectively, parallel to reduction of malondialdehyde (MDA), myeloperoxidase (MPO), glial fibrillary acidic protein (GFAP) and L-glutamate/IL-1β/TXNIB/NLRP3 axis. Moreover, biochanin A suppressed neuronal damage by reducing the astrocytes' activation and significantly attenuated the PTZ-induced increase in LC3 levels by 55.5%. Furthermore, molecular docking findings revealed that BIOCHANIN A has a higher affinity for phosphoinositide 3-kinase (PI3k), threonine kinase2 (AKT2), and mammalian target of rapamycin complex 1 (mTORC1) indicating the neuroprotective and anti-epileptic characteristics of biochanin A in the brain tissue of PTZ-kindled mice.

Combination of metformin and sub-therapeutic dose of valproic acid prevent valproic acid-induced toxicity in animal model of epilepsy

Valproic acid (VPA) is one of the most prescribed drugs for epilepsy. Extended use of VPA not only induces hepatotoxicity but also impairs the cognitive functions. Metformin has been reported to prevent epileptogenesis and enhance memory. To counter the VPA-induced adverse events, it is hypothesized that combination of sub-therapeutic dose of VPA with metformin may attenuate the toxicity stemming from the therapeutic dose of VPA. Pentylenetetrazole (PTZ)-induced kindling model of epilepsy in mice was used to assess the combined effects of sub-therapeutic dose of VPA (100 mg/kg) and metformin (200 mg/kg). The memory performance was analyzed by passive avoidance test, while alkaline comet assay was used to determine genotoxicity. Histopathological examination and serum biochemical analysis was performed to determine hepatotoxicity. Results showed that combination dose of VPA with metformin reduced seizure scores. VPA (300 mg/kg) administered as a single agent did not enhance memory impairment caused by PTZ, however, combination of sub-therapeutic dose of VPA with metformin enhanced memory function. Furthermore, in alkaline comet assay, combination therapy demonstrated reduced genotoxicity compared to the VPA 300 mg/kg. Histopathological examination of liver and analysis of serum hepatic enzymes revealed that combination therapy (VPA + metformin) reversed the toxicity as seen in case of PTZ or VPA (300 mg/kg) treated animals with no other treatment given. Based on the study data, it is concluded that the combination of sub-therapeutic dose of VPA with metformin might be used for epileptic seizures. This will prevent the hepatotoxicity and enhanced memory functions as compared to the VPA given as a single agent therapy.

Dietary Zinc Differentially Regulates the Effects of the GPR39 Receptor Agonist, TC-G 1008, in the Maximal Electroshock Seizure Test and Pentylenetetrazole-Kindling Model of Epilepsy.

The G-protein coupled receptor 39 (GPR39) is gaining increasing attention as a target for future drugs, yet there are gaps in the understanding of its pharmacology. Zinc is an endogenous agonist or an allosteric modulator, while TC-G 1008 is a synthetic, small molecule agonist. Zinc is also a positive allosteric modulator for the activity of TC-G 1008 at GPR39. Activation of GPR39 by TC-G 1008 facilitated the development of epileptogenesis in the pentylenetetrazole (PTZ)-induced kindling model of epilepsy. Congruently, TC-G 1008 decreased the seizure threshold in the maximal electroshock seizure threshold (MEST) test. Here, we investigated the effects of TC-G 1008 under the condition of zinc deficiency. Mice were fed a zinc-adequate diet (ZnA, 50 mg Zn/kg) or a zinc-deficient diet (ZnD, 3 mg Zn/kg) for 4 weeks. Following 4 weeks of dietary zinc restriction, TC-G 1008 was administered as a single dose and the MEST test was performed. Additional groups of mice began the PTZ-kindling model during which TC-G 1008 was administered repeatedly and the diet was continued. TC-G 1008 administered acutely decreased the seizure threshold in the MEST test in mice fed the ZnD diet but not in mice fed the ZnA diet. TC-G 1008 administered chronically increased the maximal seizure severity and the percentage of fully kindled mice in those fed the ZnA diet, but not in mice fed the ZnD diet. Our data showed that the amount of zinc in a diet is a factor contributing to the effects of TC-G 1008 in vivo.

Neuroprotective Effect of Celastrus Paniculatus Seed Extract on Epilepsy and Epilepsy-associated Cognitive Deficits.

Cognitive deficit is one of the common comorbidity accompanying epilepsy. The present study evaluated the effect of Celastrus paniculatus seed extract on seizure severity and cognitive deficit following the pentylenetetrazole (PTZ)-induced chemical kindling model. PTZ kindling model was developed by daily administration of the sub-convulsive dose of PTZ 30 mg/kg for four weeks. After four weeks of induction, the following treatment, namely sodium valproic acid (SVA) 200 mg/kg, C. paniculatus 500 mg\kg, pergolide 2 mg/kg, C. paniculatus (250 mg\kg)+ Pergolide (1 mg/kg), and C. paniculatus (250 mg\kg)+ SVA (100 mg/kg) were administered 30 minutes prior to PTZ (30 mg/kg) injection for a period of next 14 days. Neurobehavioral parameters, including superoxide dismutase (SOD), Catalase (CAT), glutathione (GSH), and dopamine levels were assessed and the Morris water maze test (MWM) and Grip strength test (GPS) were performed. Hematoxylin & Eosin (H&E) staining of hippocampal cornu ammonis (CA1), CA2, CA3, dentate gyrus (DG), and frontal cortex was performed. C. paniculatus (500 mg/kg) alone and in combination (C. paniculatus (250 mg\ kg)+ pergolide (1 mg/kg) and C. paniculatus (250 mg\kg)+ SVA (100 mg/kg)) significantly (P<0.05) reduced the seizure score, mean latency time, and distance traveled in the MWM. However, no significant effect was seen in GPS. Biochemical analysis showed elevated antioxidant markers, namely GSH, CAT, and SOD, and also elevated dopamine levels. C. paniculatus and its combination also significantly (P<0.05) protected against neuronal loss in the hippocampus and frontal cortex evidenced by H&E staining. C. paniculatus alone and in combination with other agents may have the potential to treat epilepsy and associated cognitive deficits.