Pro-inflammatory cytokines, but not brain- and extracellular matrix-derived proteins, are increased in the plasma following electrically induced kindling of seizures.

BACKGROUND: The aim of the study was to evaluate the brain-derived proteins, extracellular matrix-derived protein and cytokines as potential peripheral biomarkers of different susceptibility to seizure development in an animal model of epilepsy evoked by chronic focal electrical stimulation of the brain. METHODS: The plasma levels of IL-1ß (interleukin 1ß), IL-6 (interleukin 6), UCH-L1 (ubiquitin C-terminal hydrolase 1), MMP-9 (matrix metalloproteinase 9), and GFAP (glial fibrillary acidic protein) were assessed. The peripheral concentrations of the selected proteins were analyzed according to the status of kindling and seizure severity parameters. In our study, increased concentrations of plasma IL-1ß and IL-6 were observed in rats subjected to hippocampal kindling compared to sham-operated rats. RESULTS: Animals that developed tonic-clonic seizures after the last stimulation had higher plasma concentrations of IL-1ß and IL-6 than sham-operated rats and rats that did not develop seizure. Elevated levels of IL-1ß and IL-6 were observed in rats that presented more severe seizures after the last five stimulations compared to sham-operated animals. A correlation between plasma IL-1ß and IL-6 concentrations was also found. On the other hand, the plasma levels of the brain-derived proteins UCH-L1, MMP-9, and GFAP were unaffected by kindling status and seizure severity parameters. CONCLUSIONS: The plasma concentrations of IL-1ß and IL-6 may have potential utility as peripheral biomarkers of immune system activation in the course of epilepsy and translational potential for future clinical use. Surprisingly, markers of cell and nerve ending damage (GFAP, UCH-L1 and MMP-9) may have limited utility.

The Therapeutic Effects of Low-Frequency Electrical Stimulations Adjunct to Sodium Valproate on Seizure and Behaviors.

INTRODUCTION: Consuming antidepressant medications induce several problems leading to the need for alternative agents for emotional disturbances. Antidepressant medications increase the seizure risk; thus, alternative treatments, like Antiepileptic Drugs (AED), might be useful for patients with epilepsy comorbid with a psychiatric disorder. The present study evaluated the behavioral effects of sodium valproate, a none effective dose in seizure treatment [100 mg/kg; Intraperitoneal (IP)] along with the application of Low-Frequency Stimulations (LFS) during CA1 hippocampal kindling. METHODS: In total, 42 male rats were randomly divided into 6 groups, including control group with intact animals handled daily (I); sham group which was subjected to the surgical process, but received no real stimulation (II); saline-kindled Kindled group (S.kindled) which were stimulated daily with the following protocol: 3 strain of 50Hz monophasic pulses of 1ms duration applied 12 times a day with the threshold intensity at intervals of 10 minutes where saline was administrated 15 min before kindling stimulations (III); saline-kindled-LFS group (K4LFS) in which saline was injected 15 min before kindling stimulations and LFS was applied daily after the termination of kindling stimulation (IV); drug-kindle group (Drug100.kindled) that underwent rapid kindling procedure daily where sodium valproate (100 mg/kg) was administrated 15 min before kindling stimulations(V), and drug-kindled-LFS (Drug100.kindled.4LFS) group in which drug and LFS were administrated respectively before and after kindling stimulations (VI). The behavioral tests were assessed using elevated plus maze, open field, and forced swim tests. RESULTS: The combination of sodium valproate (100 mg/kg) and LFS significantly decreased cumulative seizure severity compared with the kindle group. Thus, it provided a strong seizure suppressing effect. Additionally, sodium valproate and LFS increased the percentage of Open Arms (OAs) entries and the OAs exploration; they also decreased jumping from elevated plus maze test and rearing in open field test. Furthermore, there was no significant change in the OAs entries and OAs exploration percentages, jumping from apparatus, and rearing in open field in Drug100. Kindled, K4LFS, and Drug100.kindled.LFS groups, compared with the sham group. There was no significant difference in the latency to first immobility and the duration of immobility in K4LFS groups compared with the S. kindled group. In the drug-kindled group, the latency to first immobility significantly increased, and the duration of immobility decreased, compared with the S. kindled group. Besides, the latency to first immobility significantly increased, and the duration of immobility decreased in drug-kindled-LFS, compared to S. kindled group; however, the latency to first immobility was not significantly changed, compared to drug-kindled groups. CONCLUSION: Sodium valproate and LFS can modulate the function of the brain regions involved in emotional processing in epilepsy, as well as anxiety- and depressive-like behaviors. Such a combination could also decrease emotional disturbances induced by the kindling process.

The role of IL-1ß and glutamate in the effects of lipopolysaccharide on the hippocampal electrical kindling of seizures.

In our study, we used rapid electrical hippocampal kindling and in vivo microdialysis methods to assess the involvement of inflammatory mediators: lipopolysaccharide (LPS) and proinflammatory interleukin-1ß (IL-1ß) in mechanisms of epileptogenesis. We observed, that both, LPS and IL-1ß, administered into stimulated hippocampus, accelerated kindling process. LPS also increased the expression of IL-1ß in stimulated hippocampus in kindled rats. In vivo acute LPS perfusion, via a microdialysis cannula implanted into the naïve rat's hippocampus, produced an increase in extracellular glutamate release. We suppose, that particularly IL-1ß action and increased glutamate concentration may significantly contribute to LPS effects on kindling development.

Effects of hippocampal partial kindling on sensory and sensorimotor gating and methamphetamine-induced locomotion in kindling-prone and kindling-resistant rats.

The effects of hippocampal partial kindling on gating of hippocampal auditory-evoked potentials (AEPs), prepulse inhibition (PPI) to an acoustic startle response, and methamphetamine-induced locomotion were examined in selectively bred kindling-prone (Fast) and kindling-resistant (Slow) rats. Ten electrographic seizures (afterdischarges, ADs) induced by high-frequency stimulation of the hippocampal CA1 region resulted in deficits in gating of hippocampal AEP and PPI in Fast, but not Slow, rats. The increase in AD duration with kindling was similar in Fast and Slow rats. Kindling-induced changes in hippocampal AEP and PPI in Fast rats were abolished by pretest injection of CGP7930 (1mg/kg i.p.), a positive allosteric modulator of GABAB receptors. Injection of haloperidol (0.1mg/kg i.p.) daily before kindling also prevented kindling-induced changes in PPI and hippocampal AEP in Fast rats. Interestingly, methamphetamine-induced hyperlocomotion was enhanced by kindling in Slow, but not Fast, rats. However, the methamphetamine-induced hyperlocomotion in Slow rats was not suppressed by daily injection of 0.1mg/kg i.p. haloperidol before kindling, as compared with kindling without haloperidol. It is concluded that genetic disposition affected the behavioral consequences of repeated seizures. Fast rats required fewer hippocampal ADs to induce sensory (AEP) and sensorimotor (PPI) deficits, while Slow kindled rats were more sensitive to methamphetamine-induced locomotion. Dopaminergic blockade by haloperidol during kindling, or acute injection of CGP7930 before testing, attenuated some of the behavioral deficits induced by repeated hippocampal seizures, suggesting possible therapeutic strategies to treat the schizophrenic-like symptoms associated with temporal lobe epilepsy.

Improvement in hippocampal kindling analysis through computational processing data / Aprimorando a análise do modelo de kindling hipocampal com o auxílio de processamento computacional

The kindling phenomenon is classically investigated in epileptology research. The present study aims to provide further information about hippocampal kindling through computational processing data. Adult Wistar rats were implanted with dorsal hippocampal and frontal neocortical electrodes to perform the experiment. The processing data was obtained using the Spike2 and Matlab softwares. An inverse relationship between the number of "wet dog shakes" and the Racine's motor stages development was found. Moreover it was observed a significant increase in the afterdischarge (AD) duration and its frequency content. The highest frequencies were, however, only reached at the beginning of behavioral seizures. During the primary AD, fast transients (ripples) were registered in both hippocampi superimposed to slower waves. This experiment highlights the usefulness of computational processing applied to animal models of temporal lobe epilepsy and supports a relevant role of the high frequency discharges in temporal epileptogenesis.

O fenômeno de kindling é classicamente utilizado no campo da epileptologia experimental. Este trabalho objetiva aprofundar a análise do modelo kindling hipocampal através de processamento computacional. Ratos wistar adultos receberam eletrodos hipocampais dorsais e neocorticais frontais para a realização do experimento. O processamento dos dados encontrados foi realizado pelos softwares Matlab e Spike2. Encontrou-se uma relação inversa entre wet dog shakes e o desenvolvimento dos estágios motores de Racine. A duração e o conteúdo de freqüência das pós-descargas hipocampais aumentaram durante o processo, sendo observadas descargas de alta freqüência (ripples) em ambos os hipocampos durante as pós-descargas primárias, superimpostas a ondas lentas. As mais altas freqüências, entretanto, foram apenas atingidas com o início das crises epilépticas. A utilização de sistemas computacionais para a confecção e análise do modelo de epilepsia temporal é ressaltada e reforça-se a relevância do papel das altas freqüências na epileptogênese temporal.