Pentylenetetrazole-induced Seizures In Mice Research Articles

The effects of ethanolic extract of Punica granatum Peel on Pentylenetetrazole-induced seizures in mice

Introduction: Pomegranate (Punica granatum L) is an ancient and unique fruit used as antioxidant, antidiabetic, anti-obesity, and antihypertensive. Considering oxidative stress and its relationship with convulsions and the antioxidant properties of pomegranate extract, the present study aimed to investigate the effect of ethanolic extract of Punica granatum peel on pentylenetetrazole (PTZ)-induced seizures in mice. Method: In this experimental study, male rats were randomly divided into four groups (n=8 in each group, PTZ group, and three groups received 100, 200, and 400 mg/kg of the extract). Thirty minutes after peritoneal injection of different doses of the extract, PTZ (100 mg/kg) was injected, and the mice were immediately transferred to a special cage, and the seizure parameters were evaluated for 60 minutes. Results: The results implicated that treating with ethanolic extract of Punica granatum peel in different doses of 100, 200, and 400 mg/kg body weight could delay the onset of minimal clonic seizures (MCS) caused by PTZ injection. In addition, doses of 200 and 400 mg/kg delay the onset of generalized tonic-clonic seizures (GTCS) caused by PTZ injection (P<0.001 to P<0.05). The anti-seizure effects of the ethanolic extract of Punica granatum were dose-dependent; therefore, the high doses have the most prophylactic effects against MCS and GTCS seizures (P<0.05). Conclusion: The results of this study showed that treatment with ethanolic extract of Punica granatum peel has an anti-seizure effect on mice, and the highest anti-seizure effect was observed in the highest dosage

Seizure event detection using intravital two-photon calcium imaging data.

Intravital cellular calcium imaging has emerged as a powerful tool to investigate how different types of neurons interact at the microcircuit level to produce seizure activity, with newfound potential to understand epilepsy. Although many methods exist to measure seizure-related activity in traditional electrophysiology, few yet exist for calcium imaging. To demonstrate an automated algorithmic framework to detect seizure-related events using calcium imaging-including the detection of pre-ictal spike events, propagation of the seizure wavefront, and terminal spreading waves for both population-level activity and that of individual cells. We developed an algorithm for precise recruitment detection of population and individual cells during seizure-associated events, which broadly leverages averaged population activity and high-magnitude slope features to detect single-cell pre-ictal spike and seizure recruitment. We applied this method to data recorded using awake in vivo two-photon calcium imaging during pentylenetetrazol-induced seizures in mice. We demonstrate that our detected recruitment times are concordant with visually identified labels provided by an expert reviewer and are sufficiently accurate to model the spatiotemporal progression of seizure-associated traveling waves. Our algorithm enables accurate cell recruitment detection and will serve as a useful tool for researchers investigating seizure dynamics using calcium imaging.

Quercetin Exerts Anticonvulsant Effect through Mitigation of Neuroinflammatory Response in Pentylenetetrazole-induced Seizure in Mice.

Epilepsy is a chronic disease of the brain characterized by seizures. The currently available anticonvulsants only treat symptoms with serious adverse drug reactions. Therefore, there is need for new therapeutic intervention that will prevent epileptogenesis with greater therapeutic success. Quercetin (QT) is a flavonoid with known neuroprotective and anti-inflammatory properties. The study aimed to investigate its effects against pentylenetetrazole (PTZ)-induced seizures. Animals were divided into four groups (n = 10). Group 1(control) only received vehicle (10 mL/kg), group 2 received vehicle, groups 3 and 4 received QT 12.5 mg/kg and 25 mg/kg respectively. Sixty minutes after treatments, animals in groups 2 to 4 were injected with sub-convulsive dose of pentylenetetrazole (35 mg/kg, i.p.) on every alternate day (48±2h) for 21 days. The mice were observed for 30 minutes after each PTZ injection for seizure activity. Brain samples were collected for biochemical assays. Administration of PTZ caused significant increase in the intensity of seizures, neuronal degeneration and level of proinflammatory cytokines in animals compared to control. These behavioural alterations were attenuated significantly by QT (12.5 and 25 mg/kg). The PTZ-induced increase in IL-12, TNF-α and IFN-ɣ were significantly reduced by pre-treatment with the QT (12.5 and 25 mg/kg, p.o). Quercetin also reduced neuronal loss compared to control. Quercetin attenuates seizures in kindled mice and reduces neuroinflammation and neurodegeneration. This neuroprotective effect may be attributed to its ability to inhibit inflammatory mediators in the brain.

Synthesis Anticonvulsant and Cytotoxic Evaluation of Benzimidazole-Quinoline Hybrids Schiff Base Analogs

Both, benzimidazole and quinoline pharmacophores have shown various activities against cancer, tuberculosis, malaria, fungal, viral, and bacterial infections. These moieties are considered highly potent nuclei in medicinal chemistry. Thus, this research work aims to produce a synergistic effect involving both nuclei in a single structure. A Series of literature work resulted in the synthesis of novel derivatives of 2‐(1H‐1,3‐benzothiazole‐2‐yl)‐N'‐substituted quinoline‐4‐carbohydrazide 5a–q using a multiple-step reaction. Characterization of all compounds was successfully done using IR and 1HNMR, 13CNMR as well as elemental analysis. Furthermore, screening of all synthesized compounds has been shown a positive response as an anticonvulsant and antidepressant activity in mice. Compounds 5b, 5h, 5l, and 5n have shown potentially active on pentylenetetrazole-induced seizures in mice. The synthesized compounds were also screened on a panel of 60 cell lines by the National Cancer Institute according to standard screening protocol. Ten compounds were selected and evaluated via a single high dose (10−5 M). In a one-dose assay, compounds 5j and 5o showed the highest activity on leukemia (CCRF-CEM) and renal cancer (A498) with growth inhibition (GI) of 70.91 and 61.15%.

Tiagabine suppresses pentylenetetrazole-induced seizures in mice and improves behavioral and cognitive parameters by modulating BDNF/TrkB expression and neuroinflammatory markers

Tiagabine (Tia), a new-generation antiseizure drug that mimics the GABAergic signaling by inhibiting GABA transporter type-1, is the least studied molecule in chronic epilepsy models with comorbid neurobehavioral and neuroinflammatory parameters. Therefore, the current study investigated the effects of Tia in a real-time manner on electroencephalographic (EEG) activity, behavioral manifestations and mRNA expression in pentylenetetrazole (PTZ)-kindled mice. Male BALB/c mice were treated with tiagabine (0.5, 1 and 2 mg/kg) for 21 days with simultaneous PTZ (40 mg/kg) injection every other day for a total of 11 injections and monitored for seizure progression with synchronized validation through EEG recordings from cortical electrodes. The post-kindling protection from anxiety and memory deficit was verified by a battery of behavioral experiments. Isolated brains were evaluated for oxidative alterations and real-time changes in mRNA expression for BDNF/TrkB, GAT-1 and GAT-3 as well as neuroinflammatory markers. Experimental results revealed that Tia at the dose of 2 mg/kg maximally inhibited the development of full bloom seizure and reduced epileptic spike discharges from the cortex. Furthermore, Tia dose-dependently exerted the anxiolytic effects and protected from PTZ-evoked cognitive impairment. Tia reduced lipid peroxidation and increased superoxide dismutase and glutathione levels in the brain via augmentation of GABAergic modulation. PTZ-induced upregulated BDNF/TrkB signaling and pro-inflammatory cytokines were mitigated by Tia with upregulation of GAT-1 and GAT-3 transporters in whole brains. In conclusion, the observed effects of Tia might have resulted from reduced oxidative stress, BDNF/TrkB modulation and mitigated neuroinflammatory markers expression leading to reduced epileptogenesis and improved epilepsy-related neuropsychiatric effects.

Deneysel Epileptik Nöbet Modelinde, Artemisinin'in Fare Kalp ve Akciğer Dokularında Oksidatif Stress Belirteçleri Üzerine Etkisi.

Aim: The current study investigated the effects of artemisinin on the heart and lung tissue against pentylenetetrazol-induced seizures in mice. For this purpose, malondialdehyde (MDA), advanced oxidation protein products (AOPP), Catalase (CAT), glutathione (GSH), and glutathione peroxidase (GSH-Px) levels were evaluated in both tissue homogenates.Material and Method: Swiss albino male mice (n=42) were used in the experiment. Animals were divided into six groups; Control (C), pentylenetetrazol (PTZ), valproate 100 mg/kg (VPA), artemisinin 30 mg/kg (ARS)+PTZ, ARS 60 mg/kg+PTZ, ARS 120 mg/kg+PTZ. On the 26th day of the experiment, the mice were sacrificed and the samples were kept at -80 0C for biochemical analysis.Results: There were significant differences in the five biochemical parameters analyzed in heart and lung tissues. Heart and lung MDA levels of the PTZ group were found to be significantly higher than the C and ARS-60 groups (p<0.05). Heart and lung MDA levels of the PTZ group were found to be significantly higher than the C and ARS-60 groups. Likewise, heart AOPP levels decreased significantly in the VPA and ARS-60 groups compared to the PTZ group (p<0.05). There was no significant difference between the groups in terms of lung AOPP levels (p>0.05). Heart CAT and GSH levels were decreased in the PTZ group compared to the other groups. However, in terms of Lung CAT levels, the PTZ group had the highest value compared to the other groups, while it had the lowest value in terms of GSH level. The GSH-Px level did not differ significantly between the groups in heart tissue (p>0.05). The lung GSH-Px level was significantly increased in the ARS-30 group when compared to the PTZ group (p<0.05).Conclusion: Consequently ARS treatment can inhibit PTZ-induced oxidative stress in peripheral tissues. In addition, ARS may provide improvements in decreased antioxidant enzymes. ARS may contribute to the antioxidant defense system.

Protective Effect of Metformin Alone or in Combination with Valproic acid on Pentylenetetrazole-Induced Seizures in Mice

Abstract Objective: The aim of this study was to investigate the effects of metformin on pentylenetetrazole (PTZ)-induced seizures and the neuroprotective effect of metformin on neuronal damage after pentylenetetrazole administration. Material and Methods: Thirty-five (35) Male BALB-c Albino mice weighing 35-38 g were divided randomly into five groups: Control group (1), Saline+PTZ group (2), Valproic Acid (VPA, 200 mg/kg intraperitoneal-i.p.)+PTZ group (3), Metformin (200 mg/kg i.p.)+PTZ group (4), and VPA+Metformin+PTZ group (5). The PTZ (60 mg/kg, i.p.) was injected 30 min after drugs injection to induce seizures and seizure stages and behavioral scoring were evaluated. After completing procedure, brain tissues were removed and analyzed with biochemical and histopathological procedures. The hippocampal Cornu Ammonis (CA) 1, CA2, CA3 and DG (dentate gyrus) regions were histopathologically evaluated and oxidative stress markers (total antioxidant status (TAS), total oxidant status (TOS)) were measured. Results: Compare to Saline+PTZ group, metformin administration alone did not affect the onset time of the first myoclonic jerk (FMJ), but combination of VPA and metformin significantly increased FMJ onset time (p<0.05). Additionally, the treatment of metformin with or without VPA reduced the brain oxidative stress (p<0.05). Furthermore, histopathological assessment demonstrated that metformin administration and the combination of VPA and metformin decreased dark neuron formation in the hippocampal CA1, CA2, CA3, and DG areas (p<0.05). Conclusion: Metformin was found to be significantly effective in reducing epileptic seizures, brain oxidative stress, and preventing neural damage after PTZ-induced seizure

Mentha piperita Oil Exerts an Antiepileptic Effect in Pilocarpine and Pentylenetetrazol-Induced Seizures in Mice.

Introduction Epilepsy is a progressive, chronic neurological disorder characterized by recurrent seizures. Peppermint (Mentha piperita L.) (MP) is one of the most commonly ingested herbal teas or tisanes with a single component. Aim We aimed to assess the potential antiepileptic and neuroprotective features of MP essential oil (MPO) in pilocarpine (P) and pentylenetetrazol (PTZ) models of epilepsy. Methods The study used eight groups of mice to assess the anticonvulsant activity of MPO in both the P and PTZ acute models in mice. P (350 mg/kg, i.p.) was given 30 minutes after MPO (1.6, 3.2, and 6.4 ml/kg, i.p.). As a positive control group, diazepam (1 mg/kg, i.p) was used. PTZ (95 mg/kg, i.p.) was given 30 minutes after MPO (6.4 ml/kg, i.p.). The first convulsion's latency time, the number of convulsions, the latency time to death, and the percentage of deaths were calculated in all groups. Results MPO significantly (P < 0.05) increases the first convulsion's latency time and the death's latency time. Moreover, the essential oil significantly decreases the number of convulsions and reduces the mortality rate compared to the negative control group. Conclusion MPO at 3.2 and 6.4 ml/kg doses can reduce the percentage and the number of convulsions and increase the latency time of both the first convulsion and death so that it can be used as a supplement in the treatment of epilepsy.

Dual effects of anandamide in the antiepileptic activity of diazepam in pentylenetetrazole-induced seizures in mice.

The prototype endocannabinoid, anandamide activates both CB 1 and transient receptor potential vanilloid type 1 channels (TRPV1) receptor at different concentrations. At high concentrations, anandamide-mediated TRPV1 effects are opposite to its effects at low concentrations via CB 1 receptor. Thus, synaptic concentrations of anandamide govern the neuronal activity and consequently might affect the response of a drug. This study was undertaken to investigate the influence of high and low doses of anandamide on the anticonvulsant action of diazepam on the subcutaneous dose of pentylenetetrazole (PTZ) in Swiss mice weighing 20-25 g. Results revealed that intracerebroventricular administration of capsazepine (a TRPV1 antagonist: 1, 10, or 100 µg/mouse) and the low doses (10 µg/mouse) of anandamide, AM404 (anandamide transport inhibitor), or URB597 (fatty acid amide hydrolase inhibitor) augmented the anticonvulsant effect of diazepam. Conversely, higher dose of anandamide, AM404, URB597 (100 µg/mouse) as well as capsaicin (a TRPV1 agonist: 1, 10, or 100 µg/mouse) attenuated the protective effect of diazepam against PTZ-induced seizures. Thus, this study demonstrates that the effects of diazepam may be augmented by activating CB 1 receptors or dampened via TRPV1 receptors. The findings of the present study can be extrapolated to understand the use of TRPV1 blockers alone or in combination of benzodiazepines in the treatment of benzodiazepines-refractory status epilepticus, a condition associated with maladaptive trafficking of synaptic gamma-aminobutyric acid and glutamate receptors. However, potential clinical applications are needed to further support such preclinical studies.

Deficit of PKHD1L1 in the dentate gyrus increases seizure susceptibility in mice.

Epilepsy is a chronic neurological disorder featuring recurrent, unprovoked seizures, which affect more than 65 million people worldwide. Here, we discover that the PKHD1L1, which is encoded by polycystic kidney and hepatic disease1-like 1 (Pkhd1l1), wildly distributes in neurons in the central nervous system (CNS) of mice. Disruption of PKHD1L1 in the dentate gyrus region of the hippocampus leads to increased susceptibility to pentylenetetrazol-induced seizures in mice. The disturbance of PKHD1L1 leads to the overactivation of the mitogen-activated protein kinase (MAPK)/extracellular regulated kinase (ERK)-Calpain pathway, which is accompanied by remarkable degradation of cytoplasmic potassium chloride co-transporter 2 (KCC2) level together with the impaired expression and function of membrane KCC2. However, the reduction of membrane KCC2 is associated with the damaged inhibitory ability of the vital GABA receptors, which ultimately leads to the significantly increased susceptibility to epileptic seizures. Our data, thus, indicate for the first time that Pkhd1l1, a newly discovered polycystic kidney disease (PKD) association gene, is required in neurons to maintain neuronal excitability by regulation of KCC2 expression in CNS. A new mechanism of the clinical association between genetic PKD and seizures has been built, which could be a potential therapeutic target for treating PKD-related seizures.

Seratrodast, a thromboxane A2 receptor antagonist, inhibits neuronal ferroptosis by promoting GPX4 expression and suppressing JNK phosphorylation

More than 30 % of individuals with epilepsy are refractory to currently available drugs, highlighting the urgent need to develop novel candidate drugs. Accumulating evidence implicates the key role of ferroptosis in the pathophysiology of epileptic seizuresand its potential as a new drug target. Drug repurposing is a promising strategy for the rapid generation of new candidate drugs from the market drugs with new therapeutic indications, such as the best-selling drug thalidomide. Herein, we reported the discovery of Seratrodast, a market drug of thromboxane A2 receptor antagonist as a new ferroptosis inhibitor (IC50: 4.5 μmol·L-1). Seratrodast could reduce lipid ROS production, regulate the system xc-/glutathione (GSH)/glutathione peroxidase 4 (GPX4) axis, and inhibit JNK phosphorylation and p53 expression. In addition, Seratrodast elevated GPX4 expression and decreased JNK phosphorylation in pentylenetetrazole-induced seizures in mice. Seratrodast increased the latency of seizures and reduced seizure duration in pentylenetetrazole-induced seizures. Our results suggest Seratrodast might be either a ferroptosis inhibitor or a novel lead compound for further optimization of novel drug discovery.

The Anticonvulsant Activity of Thiamine, Vitamin D3, and Melatonin Combination on Pentylenetetrazole-induced Seizures in Mice

Objective:Epilepsy is a neurological disorder associated with the elevation of oxidative stress levels and alteration in vitamins and endogenous antioxidant levels. The current study was designed to evaluate the effects of Vit D3, thiamine, melatonin, and their combination on pentylenetetrazol (PTZ)-induced tonic-clonic seizures in mice along with measuring the alteration in oxidative stress markers.Methods:Male mice were randomly divided into seven groups; group I received normal saline (0.5 ml, i.p.) on the 15th day, group II received PTZ (60 mg/kg, i.p) on the 15th day, groups III-VI received diazepam (4 mg/kg), Vit D3 (6000 IU/kg, p.o.), thiamine (200 mg/kg, p.o.), and melatonin (20 mg/kg, p.o.), respectively, before PTZ (60 mg/kg, i.p.) injection on the 15th day, and group VII received the combination of Vit D3, thiamine, and melatonin, before PTZ injection on the 15th day. After PTZ injection, the seizure latency and duration were measured. The malondialdehyde (MDA) level and superoxide dismutase (SOD) activity were evaluated in mice brains.Results:In the treatment group, the seizure latency was greater and the seizure duration was lower than in the PTZ group. These agents decreased the MDA level and increased SOD activity compared to the PTZ group. The combination of these agents was more effective than each alone to increase seizure latency and reduce seizure duration.Conclusion:The combination of Vit D3, thiamine, and melatonin could improve PTZ-induced seizures more effectively than each one alone; this may be mediated by inhibiting oxidative stress and regulating various mechanisms involved in seizures.

The possible role of nitric oxide signaling and NMDA receptors in allopurinol effect on maximal electroshock- and pentylenetetrazol-induced seizures in mice

Allopurinol, a uric-acid-lowering medication, has shown its efficacy in several studies suggesting that allopurinol can be prescribed as adjunctive cure meant for intractable epilepsy. The exact mechanism of allopurinol is still unknown. This study evaluates allopurinol’s effect on seizure threshold, seizure incidence, and mortality rate in mice models. Moreover, the possible involvement of nitric oxide (NO) pathway and N-methyl-D-aspartate (NMDA) receptors are investigated.To evaluate the effect of allopurinol on seizure, we used the pentylenetetrazole (PTZ)-induced seizure along with maximal electroshock (MES)-induced seizure. To assess the underlying mechanism behind the allopurinol activity, we used nitric oxide synthase (NOS) substrate (L-arginine), NOS inhibitors (L-NAME, aminoguanidine, 7-nitroindazole), and NMDA receptor antagonist (MK-801).Intraperitoneal allopurinol administration at a dose of 50 mg/kg in mice showed a significant (p < 0.001) anti-convulsant activity in the PTZ-induced seizure. Even though pre-treatment with L-Arginine (60 mg/kg) potentiates allopurinol’s anti-convulsant effect in the PTZ-induced seizure, pre-treatment with L-NAME (10 mg/kg), aminoguanidine (100 mg/kg), and 7-nitroindazole (30 mg/kg) reversed the anti-convulsant effect of allopurinol in the PTZ-induced seizure. In addition, pre-treatment with MK-801 also decreased the anti-convulsant effect of allopurinol in the PTZ-induced seizure. While allopurinol at a dose of 50 mg/kg and 100 mg/kg did not induce protection against seizure incidence in the MES-induced seizure, it revealed a remarkable effect in reducing the mortality rate in the MES-induced seizure.Allopurinol increases the seizure threshold in PTZ-induced seizure and enhances the survival rate in MES-induced seizure. Allopurinol exerts its anti-convulsant effect, possibly through targeting NO pathway and NMDA receptors.

Synthesis and anticonvulsant activity evaluation of n-[(2,4-dichlorophenyl)methyl]-2-(2,4-dioxo-1h-quinazolin-3-yl)acetamide novel 1-benzylsubstituted derivatives

The aim. Synthesis of 1-benzylsubstituted derivatives of N-[(2,4-dichlorophenyl)methyl]-2-(2,4-dioxo-1h-quinazolin-3-yl)acetamide, and determination of affinity to GABAergic biotargets with the following anticonvulsant activity estimation using PTZ-induced seizures model in mice.&#x0D; Materials and methods. Standard organic synthesis methods were used; the structure of the synthesized compounds was proved by elemental analysis, 1H and 13C NMR spectroscopy, and LC/MS method; composition of the synthesized compounds – by elemental analysis, their individuality – by TLC and LC/MS methods. AutoDockTools-1.5.6, as well as AutoDock Vina software, was used to perform molecular docking. Anticonvulsant activity was studied using pentylenetetrazole-induced seizures in mice.&#x0D; Results. A targeted N-[(2,4-dichlorophenyl)methyl]-2-(1-(R-benzyl)-2,4-dioxo-quinazolin-3-yl)acetamides were obtained by alkylation of N-[(2,4-dichlorophenyl)methyl]-2-(2,4-dioxo-1H-quinazolin-3-yl)acetamide by corresponding 1-chloromethylbenzene in dimethylformamide environment with excess of potassium carbonate at a temperature 70-80 ˚С. Prediction of activity of 1-benzyl derivatives in the pentylenetetrazole-induced seizures in an in vivo experiment was carried out according to the obtained results of docking studies – affinity calculation for GABA receptor and GABA enzyme active sites, as well as analysis of conformational placement in them. In relation to the binding energy, the studied ligands were inferior to the reference drugs: GABA receptor positive allosteric modulators – benzamidine and diazepam, and GABA inhibitor – vigabatrin. The synthesized substances did not show anticonvulsant activity: only 2 compounds have shown a tendency to their activity manifestation according to the criterion of integral protective indicator – reduction of mortality by 17 % compared to control, as well as prolonging the time death of the animals. Comparison with the preliminary obtained results of the activity of the promising anticonvulsant N-[(2,4-dichlorophenyl)methyl] -2-(2,4-dioxo-1H-quinazolin-3-yl) acetamide N-[(2,4-dichlorophenyl)methyl]-2-(2,4-dioxo-1H-quinazolin-3-yl)acetamide made possible to prove the pharmacophore role of the cyclic amide fragment in anticonvulsant activity manifestation.&#x0D; Conclusion. The synthesis of N-[(2,4-dichlorophenyl)methyl]-2-(1-(R-benzyl)-2,4-dioxo-quinazolin-3-yl)acetamides, which have not still described in the literature, was carried out, as well as the structure of the mentioned compounds was proved. Unfortunately, the substances did not show anticonvulsant activity on the model of pentylenetetrazole-induced seizures. However, the obtained results allowed establishing the key role of the NHCO cyclic fragment on anticonvulsant activity. A positive correlation between the results of in vivo studies and in silico calculations was found – the model of pentylenetetrazole-induced seizures and docking into the active sites of PAMs GABAА receptor and enzyme inhibitor GABAАТ, which allows to recommend the given docking methodology as a tool to streamline and optimize the screening on the mentioned model

Chemical composition of Zhumeria majdae essential oil and its effects on the expression of morphine withdrawal syndrome and tolerance to the anticonvulsant effect of morphine on pentylenetetrazole-induced seizures in mice

Regarding the proven anticonvulsant effect of Zhumeria majdae essential oil (ZMEO) in previous studies we were prompted to investigate the ZMEO effects on the tolerance to the anticonvulsant effects of morphine and the morphine withdrawal syndrome. Tolerance to the morphine anticonvulsant effect was induced in mice by subcutaneous injection of 2.5 mg/kg of morphine for 4 days. Subsequent doses of ZMEO (20 mg/kg) were used to study the expression and development of morphine tolerance. Clonidine was used as the standard drug to inhibit the morphine withdrawal syndrome symptoms. To study the ZMEO effect on withdrawal syndrome, mice received appropriate morphine values for 4 days and on the fifth day, 60 min before administration of naloxone. The effective dose of ZMEO was determined and the number of jumps, stands and changes in the dry stool weight, as symptoms of withdrawal syndrome were evaluated. The dose of 20 mg/kg of ZMEO decreased the tolerance in development and expression groups significantly. Counting the number of jumping, standing and defecation were assessed 30 min after morphine and 1 h after the vehicle and clonidine. The dose of 40 mg/kg ZMEO decreased all the signs of withdrawal syndrome significantly. ZMEO was analyzed by GC/MS and linalool (53.1%) and camphor (23.8%) were characterized as the main components. The results suggest that ZMEO possesses constituent(s) that have activity against tolerance to the anticonvulsant effects of morphine and the morphine withdrawal symptoms.

Anchusa italica Retz. Hydro-Alcoholic Extract through Attenuation of Oxidative Stress Exerts an Anticonvulsant Effect on the Pentylenetetrazole-Induced Seizure in Mice

AbstractIntroduction It has been shown that oxidative stress is involved in the pathophysiology of seizure. Current anticonvulsants have side effects, thus further studies are needed to find new agents with low side effects. Anchusa italica Retz. has been suggested to have antioxidant and neuroprotective effects. The present study aimed to determine the anticonvulsant effects of A. italica hydro-alcoholic extract on pentylenetetrazole (PTZ)-induced seizures in mice focusing on its possible antioxidative stress properties.Materials and Methods Sixty mice were randomly divided into six groups. The intervention groups received the A. italica extract at the doses of 50, 100, and 200 mg/kg, 30 minutes before the injection of PTZ, whereas the positive control group received phenobarbital. The seizure threshold was then recorded.Results A. italica extract significantly increased the seizure threshold. The extract significantly increased serum and prefrontal cortex total antioxidant capacity compared with the control group. The A. italica extract significantly reduced serum and prefrontal cortex malondialdehyde and nitrite levels compared with the control group.Conclusions The results showed the anticonvulsant effects of A. italica extracts on the PTZ-induced seizure in mice is partially due to the attenuation of oxidative stress.

Effect of 70% Ethanol Extract and its Solvent Fractions of Artemisia afra (Jacq. Ex Willd.) against Pentylenetetrazole-Induced Seizure in Mice.

Introduction Artemisia afra (Jacq. ex Willd.), commonly called African wormwood, is a highly aromatic perennial herb and a well-known medicinal plant, claimed to be effective and safe in the treatment of epilepsy. The whole-plant extract is traditionally used as an antiepileptic agent in Ethiopia. Aim of the Study. The aim of this study was, therefore, to evaluate the anticonvulsant effect of the hydroethanolic extract and solvent fractions of A. afra whole part in mice. Materials and Methods The effects of A. afra hydroethanolic extract and its solvent fractions were evaluated against pentylenetetrazole- (PTZ-) induced convulsions in mice. The onset and duration of PTZ-induced convulsions were determined with hydroethanolic A. afra extract and its solvent fractions. Data were analyzed using a one-way analysis of variance (ANOVA) followed by post hoc Tukey's multiple comparisons test. p < 0.05 was considered statistically significant. Results The hydroethanolic extract of A. afra, with all the three doses of 250, 500, and 1000 mg/kg, showed a significant delay (504.833 ± 62.835∗ s; p < 0.05∗; 551.833 ± 47.69∗∗ s; p < 0.01∗∗; and 808.333 ± 64.8∗∗∗s; p < 0.001∗∗∗, respectively) in the mean onset of convulsion and a decrease (17.000 ± 1.88∗∗∗ s, p < 0.05∗; 13.000 ± 1.8∗∗ s, p < 0.01∗∗; and 7.833 ± 1.07∗∗∗ s, p < 0.001, respectively) in the mean duration of convulsion against PTZ-induced convulsion in a dose-dependent manner compared to the control (92.833 ± 13.006 s; 34.167 ± 3.683 s), and its anticonvulsant activity was significantly less compared to that of diazepam (1001.167 ± 68.430 s; 4.500 ± 0.619 s). The solvent fractions, however, did not show anticonvulsant activity against PTZ-induced convulsion. Conclusion Crude extract of A. afra has an anticonvulsant effect in mice. This might be attributed to the synergistic effects of two or more active ingredients present in the herb.

Порівняльне дослідження антиконвульсивного потенціалу серцевих ґлікозидів за умов пентилентетразолових судом у мишей

Aim. The research provides a comparative analysis of the possible anticonvulsant action of different cardiac glycosides, namely: digoxin, lanatoside C, strophanthin G and corglycone. In addition, it detrmines the leading medication among the abovementioned ones by dose-dependence of its anticonvulsant action. Material and Methods. The research was performed on 66 random-bred albino male mice. The anticonvulsant effect of cardiac glycosides was studied in a baseline model of pentylenetetrazole-induced seizures. The first series of experiment evaluated the effect of cardiac glycosides on the course of model seizures in comparable doses of approximately 1/10 LD50 for the corresponding drug: digoxin, lanatoside C and strophanthin G - at a dose of 0.8 mg/kg; corglycone - at a dose of 1 mg/kg. The second series of experiments used the drug-leader, which was digoxin, in a wide dose range from 0.2 to 1.6 mg/kg. Digoxin, strophanthin G and corglycone were administered subcutaneously for 15 minutes, lanatoside C - intragastrically for 30 minutes before the induction of experimental seizures. Convulsive agent - pentylenetetrazole in the form of an aqueous solution was administered to animals subcutaneously at a dose of 80 mg/kg. Results and Discussion. Digoxin at a dose of 0.8 mg/kg under conditions of pentylenetetrazole-induced seizures shows a pronounced anticonvulsant activity: it is the only one among the studied cardiac glycosides that probably reduces lethality. In addition, digoxin prolongs the latency period of the first attacks, and reduces the number of clonic-tonic paroxysms in 1 mouse. Moderate anticonvulsant properties of lanatoside C were found both by a statistically significant decrease in the number of clonic-tonic seizures in 1 mouse, and by a significant reduction in the duration of the convulsive period. Although strophanthin G is unlikely to affect lethality, it moderately reduces the severity of pentylenetetrazole-induced seizures in mice, as evidenced by a statistically significant prolongation of the latency period of the first seizures, as well as a decrease in the number of clonic-tonic seizures in 1 mouse and the duration of seizures. Prophylactic administration of corglycone only prolongs the latency period of seizures and significantly reduces the number of clonic-tonic seizures in 1 mouse. The results of the dose-dependence study of digoxin anticonvulsant action show a clear anticonvulsant potential of this cardiac glycoside in a wide range of doses - from 0.2 to 1.6 mg/kg - with a maximum effect at a dose of 0.8 mg/kg. Conclusions. It was found that cardiac glycosides have a different severity of anticonvulsant effect: the most powerful anticonvulsant effect is due to digoxin, lanatoside C and strophanthin G have moderate properties, and the least pronounced effect is characteristic to corglycone. In addition, it was determined that digoxin exhibits anticonvulsant properties in a wide range of doses, and has the most pronounced anticonvulsant effect at a dose of 0.8 mg/kg. The obtained results substantiate the expediency of further in-depth study of digoxin as an anticonvulsant medicine.

Behavioural and anticonvulsant properties of methanol extract of the stem bark of Stereospermum kunthianum Cham in mice

Decoctions of Stereospermum kunthianum Cham are used in Nigeria's folk medicine for the management of epilepsy and their efficacies are commonly applauded among the rural communities of northern Nigeria. The aim of the study is to investigate the behavioural and anticonvulsant properties of the standardized methanol extract of the stem bark of S. kunthianum, in order to scientifically establish its potential values in the management of convulsive and anxiety disorders. The oral median lethal dose (LD50) is determined in mice. The anticonvulsant effect of S. kunthianum was evaluated on pentylenetetrazole induced-seizure in mice and phenobarbitone induced-sleep was used to examine the sedative effect of the extract, marble burying test was performed to investigate the anxiolytic effect. The oral LD50 of the extract was assessed to be greater than 5000 mg/kg. S. kunthianum significantly extend the onset of pentylenetetrazole induced seizure and reduced the episodes as well. It shortened the onset and prolonged the duration of Phenobarbitone-induced sleep, it also significantly decreased the number of marbles buried by the mice, signifyinganxiolytic effect. Our study offers scientific proof that the extract may possess psychoactive principles with potential anticonvulsant and anxiolytic properties.&#x0D; Keywords: Anticonvulsant, Anxiolytic, Sedative, Mice, Stereospermum kunthianum

Susceptibility of pentylenetetrazole-induced seizures in mice with Cereblon gene knockout

Epilepsy is a neurological disorder characterized by unpredictable seizures, which are bursts of electrical activity that tempo-rarily affect the brain. Cereblon (CRBN), a DCAFs (DDB1 and CUL4associated factors), is a well-established protein associated with human mental retardation. Being a substrate receptor of the cullin-RING E3 ubiquitin ligase (CRL) 4 complex, CRBN mediates ubiquitination of several substrates and conducts multiple biological processes. In the central nervous system, the large-conductance Ca2+-activated K+ (BKCa) channel, which is the substrate of CRBN, is an important regulator of epilepsy. Despite the functional role and importance of CRBN in the brain, di-rect injection of pentylenetetrazole (PTZ) to induce seizures in CRBN knock-out mice has not been challenged. In this study, we investigated the effect of PTZ in CRBN knock-out mice. Here, we demonstrate that, compared with WT mice, CRBN knock-out mice do not show the intensification of seizures by PTZ induction. Moreover, electroencephalography recordings were also performed in the brains of both WT and CRBN knockout mice to identify the absence of significant differences in the pattern of seizure activities. Consistently, immunoblot analysis for validating the protein level of the CRL4 complex containing CRBN (CRL4Crbn) in the mouse brain was carried out. Taken together, we found that the deficiency of CRBN does not affect PTZ-induced seizure.