Investigation of the Effect of Anakinra on Pentylenetetrazole Induced Epileptic Seizures in Mice

Abstract

Purpose: Disruption of the balance between gamma aminobutyric acid (GABA) and glutamate in the brain is one of the important factors contributing to seizure formation and epileptogenesis. The aim of this study is to determine whether anakinra pretreatment has an effect on cortical and hippocampal GABA and glutamate levels in a seizure model induced with pentylenetetrazole (PTZ).

Materials and Methods: In the study, 18 BALB-c type mice were divided into 3 groups as Control, PTZ, and Anakinra groups. An injection of 60 mg/kg PTZ was administered intraperitoneally to mice to induce seizures. Anakinra group was administered intraperitoneally (100 mg/kg) 30 minutes before PTZ injection. The cortical and hippocampal GABA and glutamate levels of the animals were measured using the Enzyme Linked Immunosorbent Assay (ELISA) method.

Results: PTZ-induced seizures led to a decrease in the hippocampal GABA level (p<0.05), while the glutamate level increased (p<0.001). The treatment of PTZ also increased the level of cortical glutamate (p<0.05). The pretreatment with anakinra led to a decrease in the hippocampal glutamate level (p<0.001), but increased GABA level (p<0.01). In addition, the pretreatment with anakinra cause to decrease the level of cortical glutamate (p<0.05), while the GABA level increased (p<0.001).

Conclusion: PTZ-induced seizures cause a decrease in GABA levels and an increase in glutamate levels in the brain. Anakinra treatment prior to PTZ injection ameliorates the GABA/Glutamate imbalance that has shifted to the excitation side in the brains of mice. 

Keywords

• GABA
• Glutamat
• PTZ
• Anakinra

Farelerde Pentilentetrazol ile Oluşturulan Epileptik Nöbetler Üzerine Anakinra’nın Etkisinin Araştırılması

Abstract

Amaç: Beyinde gama aminobutirik asit (GABA) ve glutamat arasındaki dengenin bozulması, nöbet oluşumuna ve epileptogeneze katkı sağlayan önemli faktörlerden biridir. Bu çalışmanın amacı, anakinra ön tedavisinin pentilentetrazol (PTZ) ile oluşturulan nöbet modelinde kortikal ve hipokampal GABA ve glutamat seviyeleri üzerine etkisinin olup olmadığını belirlemektir.

Araçlar ve Yöntem: Çalışmada 18 adet BALB-c türü fare Kontrol, PTZ ve Anakinra grupları şeklinde 3 gruba ayrıldı. Nöbetleri başlatmak için 60 mg/kg dozda PTZ enjeksiyonu farelere intraperitonal olarak uygulandı. Anakinra grubuna PTZ enjeksiyonundan 30 dakika önce intraperitonal olarak anakinra (100 mg/kg) uygulandı. Hayvanların kortikal ve hipokampal GABA ve glutamat düzeyleri Enzyme Linked Immunosorbent Assay (ELISA) yöntemi kullanılarak ölçüldü.

Bulgular: PTZ'nin neden olduğu nöbetler, hipokampusta glutamat seviyesini artırırken (p<0.001), GABA düzeyinde azalmaya yol açtı (p<0.05). PTZ tedavisi kortikal glutamat seviyesini arttırdı (p<0.05). Anakinra ile ön tedavi hipokampusta glutamat düzeyini azaltırken (p<0.001), GABA düzeyinde artışa yol açtı (p<0.01). Ayrıca anakinra ön tedavisi kortikal glutamat seviyesini azaltırken (p<0.05), GABA düzeyinde artışla sonuçlandı (p<0.001).

Sonuç: PTZ'nin neden olduğu nöbetler beyinde GABA düzeyinin azalmasına ve glutamat seviyesinin artışına neden olmaktadır. PTZ enjeksiyonu öncesinde anakinra tedavisi farelerin beyinlerinde uyarılma lehine bozulmuş GABA/Glutamat dengesizliğini iyileştir-mektedir.

Keywords

• GABA
• Glutamat
• PTZ
• Anakinra

References

1. 1. Taskıran AS, Ergul M, Gunes H, Ozturk A, Sahin B, Ozdemir E. The Effects of Proton Pump Inhibitors (Pantoprazole) on Pentylenetetrazole-Induced Epileptic Seizures in Rats and Neurotoxicity in the SH-SY5Y Human Neuroblastoma Cell Line. Cell Mol Neurobiol. 2020;41(1):173-183.
2. 2. Sharma R, Leung WL, Zamani A, O'Brien TJ, Casillas Espinosa PM, Semple BD. Neuroinflammation in Post-Traumatic Epilepsy: Pathophysiology and Tractable Therapeutic Targets. Brain Sci. 2019;9(11):318.
3. 3. Löscher W, Potschka H, Sisodiya SM, Vezzani A. Drug Resistance in Epilepsy: Clinical Impact, Potential Mechanisms, and New Innovative Treatment Options. Pharmacol Rev. 2020;72(3):606-638.
4. 4. Narayanan J, Simon KC, Choi J, et al. Factors Affecting Cognition and Depression in Adult Patients with Epilepsy. J Epilepsy Res. 2019;9(2):103-110.
5. 5. Vezzani A, French J, Bartfai T, Baram TZ. The role of inflammation in epilepsy. Nat Rev Neurol. 2011;7(1):31-40.
6. 6. De Vries EE, van den Munckhof B, Braun KP, van Royen-Kerkhof A, de Jager W, Jansen FE. Inflammatory mediators in human epilepsy: A systematic review and meta-analysis. Neurosci Biobehav Rev. 2016;63(4):177-190.
7. 7. Vezzani A, Balosso S, Ravizza T. Neuroinflammatory pathways as treatment targets and biomarkers in epilepsy. Nat Rev Neurol. 2019;15(8):459-472.
8. 8. Maroso M, Balosso S, Ravizza T, Liu J, Bianchi ME, Vezzani A. Interleukin-1 type 1 receptor/Toll-like receptor signalling in epilepsy: the importance of IL-1beta and high-mobility group box 1. J Intern Med. 2011;270(4):319-326.

9. 9. Vezzani A, Maroso M, Balosso S, Sanchez MA, Bartfai T. IL-1 receptor/Toll-like receptor signaling in infection, inflammation, stress and neurodegeneration couples hyperexcitability and seizures. Brain Behav Immun. 2011;25(7):1281-1289.
10. 10. Dinarello CA, Simon A, van der Meer JW. Treating inflammation by blocking interleukin-1 in a broad spectrum of diseases. Nat Rev Drug Discov. 2012;11(8):633-652.
11. 11. Pascual V, Allantaz F, Arce E, Punaro M, Banchereau J. Role of interleukin-1 (IL-1) in the pathogenesis of systemic onset juvenile idiopathic arthritis and clinical response to IL-1 blockade. J Exp Med. 2005;201(9):1479-1486.
12. 12. Vezzani A, Moneta D, Conti M, et al. Powerful anticonvulsant action of IL-1 receptor antagonist on intracerebral injection and astrocytic overexpression in mice. Proc Natl Acad Sci U S A. 2000;97(21):11534-11539.
13. 13. Yan Veenendaal TM, Backes WH, Tse DHY, et al. High field imaging of large-scale neurotransmitter networks: Proof of concept and initial application to epilepsy. Neuroimage Clin. 2018;19:47-55.
14. 14. Dyomina AV, Zubareva OE, Smolensky IV, et al. Anakinra Reduces Epileptogenesis, Provides Neuroprotection, and Attenuates Behavioral Impairments in Rats in the Lithium-Pilocarpine Model of Epilepsy. Pharmaceuticals (Basel). 2020;13(11):340.
15. 15. Taştemur Y, Gumus E, Ergül M, et al. Positive effects of angiotensin-converting enzyme (ACE) inhibitor, captopril, on pentylenetetrazole-induced epileptic seizures in mice. Trop J Pharm Res. 2020;19(3):637-643.
16. 16. Ernst O, Zor T. Linearization of the bradford protein assay. J Vis Exp. 2010;38:1918.
17. 17. Mendiola AS, Cardona AE. The IL-1β phenomena in neuroinflammatory diseases. J Neural Transm (Vienna). 2018;125(5):781-795.
18. 18. Rothwell NJ, Hopkins SJ. Cytokines and the nervous system II: Actions and mechanisms of action. Trends Neurosci. 1995;18(3):130-136.
19. 19. Patel RR, Khom S, Steinman MQ, et al. IL-1β expression is increased and regulates GABA transmission following chronic ethanol in mouse central amygdala. Brain Behav Immun. 2019;75(1):208-219.
20. 20. De Simoni MG, Perego C, Ravizza T, et al. Inflammatory cytokines and related genes are induced in the rat hippocampus by limbic status epilepticus. Eur J Neurosci. 2000;12(7):2623-2633.
21. 21. Pascual V, Allantaz F, Arce E, Punaro M, Banchereau J. Role of interleukin-1 (IL-1) in the pathogenesis of systemic onset juvenile idiopathic arthritis and clinical response to IL-1 blockade. J Exp Med. 2005;201(9):1479-1486.
22. 22. Doughty PT, Hossain I, Gong C, et al. Novel microwire-based biosensor probe for simultaneous real-time measurement of glutamate and GABA dynamics in vitro and in vivo. Sci Rep. 2020;10(1):12777.
23. 23. Xiang J, Jiang Y. Antiepileptic potential of matrine via regulation the levels of gamma-aminobutyric acid and glutamic acid in the brain. Int J Mol Sci. 2013;14(12):23751-23761.
24. 24. Walsh LA, Li M, Zhao TJ, Chiu TH, Rosenberg HC. Acute pentylenetetrazol injection reduces rat GABAA receptor mRNA levels and GABA stimulation of benzodiazepine binding with No effect on benzodiazepine binding site density. J Pharmacol Exp Ther. 1999;289(3):1626-1633.
25. 25. Koshal P, Kumar P. Neurochemical modulation involved in the beneficial effect of liraglutide, GLP-1 agonist on PTZ kindling epilepsy-induced comorbidities in mice. Mol Cell Biochem. 2016;415(1-2):77-87.
26. 26. Lacoste L, Bartolucci S, Lapointe J. Pentylenetetrazole inhibits glutamate dehydrogenase and aspartate aminotransferase, and stimulates GABA aminotransferase in homogenates from rat cerebral cortex. Can J Physiol Pharmacol. 1988;66(9):1135-1138.
27. 27. Wang S, Cheng Q, Malik S, Yang J. Interleukin-1beta inhibits gamma-aminobutyric acid type A (GABA(A)) receptor current in cultured hippocampal neurons. J Pharmacol Exp Ther. 2000;292(2):497-504.
28. 28. Zhang R, Yamada J, Hayashi Y, Wu Z, Koyama S, Nakanishi H. Inhibition of NMDA-induced outward currents by interleukin-1beta in hippocampal neurons. Biochem Biophys Res Commun. 2008;372(4):816-820.
29. 29. El-Missiry MA, Othman AI, Amer MA, Sedki M, Ali SM, El-Sherbiny IM. Nanoformulated ellagic acid ameliorates pentylenetetrazol-induced experimental epileptic seizures by modulating oxidative stress, inflammatory cytokines and apoptosis in the brains of male mice. Metab Brain Dis. 2020;35(2):385-399.
30. 30. Kola PK, Akula A, Nissankara Rao LS, Danduga RCSR, Mohammad A, Ineedi S. Naringin in a combined therapy with phenytoin on pentylenetetrazole-induced kindling in rats. Epilepsy Behav. 2018;89(12):159-168.
31. 31. Dorandeu F, Barbier L, Dhote F, Testylier G, Carpentier P. Ketamine combinations for the field treatment of soman-induced self-sustaining status epilepticus. Review of current data and perspectives. Chem Biol Interact. 2013;203(1):154-159.