Journal of Sport and Exercise Physiology

Journal of Sport and Exercise Physiology

The effect of eight weeks of moderate-intensity aerobic exercise on the expression of IRAK4, MRP8, and TNF-α proteins in the hippocampus of epileptic rats

Document Type : Original Article

Authors
Mohammadreza Kordi 1
Khadijeh Ebadizal 2
Siroos Choobineh 1
Amir Hossein Saffar Kohneh Quchan 1
1 Department of Exercise Physiology, Faculty of Sport Sciences and Health, University of Tehran, Tehran, Iran
2 Exercise Physiology, Aras International Campus, University of Tehran, Tabriz, Iran
10.48308/joeppa.2025.240682.1377
Abstract
Background and Purpose: Epilepsy is a chronic neurological disorder and the fourth most common neurological disorder worldwide, in which an individual experiences two or more seizures without a specific cause during their lifetime. Inflammation is one of the important factors in exacerbating epilepsy, increasing neuronal excitability, and triggering seizure attacks. The aim of this study was to investigate the effect of eight weeks of moderate-intensity aerobic exercise on the levels of MRP8, TNF-α, and IRAK4 proteins in the hippocampus of epileptic rats.


Materials and Methods: In this regard, 32 male Wistar rats were divided into four groups, including Control (n=8), Sham (n=8), Epilepsy (n=8), and Epilepsy + Moderate-intensity aerobic exercise (n=8). After one week of adaptation to the laboratory environment, epilepsy was induced via intrahippocampal injection of one microgram of kainic acid using the stereotaxic method. Epilepsy induction was confirmed by using the Racine behavioral scale . The exercise group performed moderate-intensity aerobic exercises for 40 minutes per session, five days a week, for eight weeks. Fourty eight hours after the last training session, the rats were anesthetized, and their hippocampal tissue was extracted. The levels of Myeloid-related protein 8 (MRP8), Tumor Necrosis Factor alpha (TNF-α), and Interleukin-1 Receptor-Associated Kinase 4 (IRAK4) proteins were measured using the Western blot technique. To examine the differences in MRP8, IRAK4, and TNF-α levels between groups, one-way ANOVA and Tukey’s post-hoc test were used. Furthermore, to evaluate differences in seizure frequency, an independent t-test was used.


Results: The results showed that epilepsy induction significantly increased the expression of IRAK4 (p
Keywords
Inflammation
Seizures
Regular exercise
Central nervous system
Subjects
1. Fisher RS, Acevedo C, Arzimanoglou A, Bogacz A, Cross JH, Elger CE, et al. ILAE official report: a practical clinical definition of epilepsy. Epilepsia. 2014;55(4):475-82. https://doi.org/10.1111/epi.12550 2. Pitkänen A, Lukasiuk K. Mechanisms of epileptogenesis and potential treatment targets. Lancet Neurol. 2011;10(2):173-86. https://doi.org/10.1016/S1474-4422(10)70310-0 3. Hollis A, Lukens JR. Role of inflammasomes and neuroinflammation in epilepsy. Immunol Rev. 2025;329(1):e13421. https://doi.org/10.1111/imr.13421 4. Kong H, Yin F, He F, Omran A, Li L, Wu T, et al. The Effect of miR-132, miR-146a, and miR-155 on MRP8/TLR4-Induced Astrocyte-Related Inflammation. J Mol Neurosci. 2015;57(1):28-37. https://doi.org/10.1007/s12031-015-0574-x 5. Liu H, Roy M, Tian FF. MicroRNA-based therapy: a new dimension in epilepsy treatment. Int J Neurosci. 2013;123(9):617-22. https://doi.org/10.3109/00207454.2013.789513 6. Gan N, Yang L, Omran A, Peng J, Wu L, He F, et al. Myoloid-related protein 8, an endogenous ligand of Toll-like receptor 4, is involved in epileptogenesis of mesial temporal lobe epilepsy via activation of the nuclear factor-κB pathway in astrocytes. Mol Neurobiol. 2014;49(1):337-51. https://doi.org/10.1007/s12035-013-8522-7 7. Zhang S, Chen F, Zhai F, Liang S. Role of HMGB1/TLR4 and IL-1β/IL-1R1 Signaling Pathways in Epilepsy. Front Neurol. 2022;13:904225. https://doi.org/10.3389/fneur.2022.904225 8. Cavalcante BRR, Improta-Caria AC, Melo VH, De Sousa RAL. Exercise-linked consequences on epilepsy. Epilepsy Behav. 2021;121(Pt A):108079. https://doi.org/10.1016/j.yebeh.2021.108079 9. Ribarič S. Physical exercise, a potential non-pharmacological intervention for attenuating neuroinflammation and cognitive decline in Alzheimer’s disease patients. International journal of molecular sciences. 2022;23(6):3245. https://doi.org/10.3390/ijms23063245 10. Häfele CA, Freitas MP, da Silva MC, Rombaldi AJ. Are physical activity levels associated with better health outcomes in people with epilepsy? Epilepsy & Behavior. 2017;72:28-34. https://doi.org/10.1016/j.yebeh.2017.04.038 11. Allendorfer JB, Brokamp GA, Nenert R, Szaflarski JP, Morgan CJ, Tuggle SC, et al. A pilot study of combined endurance and resistance exercise rehabilitation for verbal memory and functional connectivity improvement in epilepsy. Epilepsy & Behavior. 2019;96:44-56. https://doi.org/10.1016/j.yebeh.2019.04.020 12. Feter N, Alt R, Häfele CA, da Silva MC, Rombaldi AJ. Effect of combined physical training on cognitive function in people with epilepsy: results from a randomized controlled trial. Epilepsia. 2020;61(8):1649-58. https://doi.org/10.1111/epi.16588 13. McAuley JW, Long L, Heise J, Kirby T, Buckworth J, Pitt C, et al. A prospective evaluation of the effects of a 12-week outpatient exercise program on clinical and behavioral outcomes in patients with epilepsy. Epilepsy & Behavior. 2001;2(6):592-600. https://doi.org/10.1006/ebeh.2001.0271 14. Shishmanova-Doseva M, Georgieva K, Uzunova Y, Ioanidu L, Atanasova M, Nenchovska Z, Tchekalarova J. Pre- and Post-Endurance Training Mitigates the Rat Pilocarpine-Induced Status Epilepticus and Epileptogenesis-Associated Deleterious Consequences. Int J Mol Sci. 2022;23(21). https://doi.org/10.3390/ijms232113188 15. de Lima Rosa G, Guzzo EFM, Nunes SEB, Padilha RB, Domingues AM, Barbosa BB, et al. Aerobic exercise, alone or combined with an anti-inflammatory drug, reduces the severity of epileptic seizures and levels of central pro-inflammatory cytokines in an animal model of epileptic seizures. Epilepsy Res. 2022;186:107018. https://doi.org/10.1016/j.eplepsyres.2022.107018 16. Allendorfer JB, Brokamp GA, Nenert R, Szaflarski JP, Morgan CJ, Tuggle SC, et al. A pilot study of combined endurance and resistance exercise rehabilitation for verbal memory and functional connectivity improvement in epilepsy. Epilepsy Behav. 2019;96:44-56. https://doi.org/10.1016/j.yebeh.2019.04.020 17. Tchekalarova J, Shishmanova M, Atanasova D, Stefanova M, Alova L, Lazarov N, Georgieva K. Effect of endurance training on seizure susceptibility, behavioral changes and neuronal damage after kainate-induced status epilepticus in spontaneously hypertensive rats. Brain Res. 2015;1625:39-53. https://doi.org/10.1016/j.brainres.2015.08.022 18. Gomes da Silva S, de Almeida AA, Silva Araújo BH, Scorza FA, Cavalheiro EA, Arida RM. Early physical exercise and seizure susceptibility later in life. Int J Dev Neurosci. 2011;29(8):861-5. https://doi.org/10.1016/j.ijdevneu.2011.07.011 19. Cerqueira É, Marinho DA, Neiva HP, Lourenço O. Inflammatory effects of high and moderate intensity exercise—a systematic review. Frontiers in physiology. 2020;10:489354. https://doi.org/10.3389/fphys.2019.01550 20. Paolucci EM, Loukov D, Bowdish DM, Heisz JJ. Exercise reduces depression and inflammation but intensity matters. Biological psychology. 2018;133:79-84. https://doi.org/10.1016/j.biopsycho.2018.01.015 21. Racine RJ. Modification of seizure activity by electrical stimulation. II. Motor seizure. Electroencephalogr Clin Neurophysiol. 1972;32(3):281-94. https://doi.org/10.1016/0013-4694(72)90177-0 22. Arida RM, Sanabria ERG, da Silva AC, Faria LC, Scorza FA, Cavalheiro EA. Physical training reverts hippocampal electrophysiological changes in rats submitted to the pilocarpine model of epilepsy. Physiology & behavior. 2004;83(1):165-71. https://doi.org/10.1016/j.physbeh.2004.08.008 23. Michev A, Orsini A, Santi V, Bassanese F, Veraldi D, Brambilla I, et al. An Overview of The Role of Tumor Necrosis Factor-Alpha in Epileptogenesis and Its Terapeutic Implications. Acta Biomed. 2022;92(S4):e2021418. https://doi.org/ 10.23750/abm.v92iS4.12667 24. Jia Y, Tang L, Yao Y, Zhuo L, Qu D, Chen X, et al. Low-intensity exercise combined with sodium valproate attenuates kainic acid-induced seizures and associated co-morbidities by inhibiting NF-κB signaling in mice. Front Neurol. 2022;13:993405. https://doi.org/10.3389/fneur.2022.993405 25. Gill R, Tsung A, Billiar T. Linking oxidative stress to inflammation: Toll-like receptors. Free Radical Biology and Medicine. 2010;48(9):1121-32. https://doi.org/10.1016/j.freeradbiomed.2010.01.006 26. Zhao T, Zhang X, Cui X, Su S, Li L, Chen Y, et al. Inhibiting the IRAK4/NF-κB/NLRP3 signaling pathway can reduce pyroptosis in hippocampal neurons and seizure episodes in epilepsy. Experimental Neurology. 2024;377:114794. https://doi.org/10.1016/j.expneurol.2024.114794 27. Ogunsuyi O, Aro P, Umar H, Oboh G. Administration of curcumin plus treadmill exercise modulate some neuronal enzyme activities, redox markers and BDNF mRNA expression in pilocarpine-induced epileptic seizure in rats. Neurochemical Journal. 2023;17(3):467-76. https://doi.org/10.1134/S1819712423030145 28. Holmes PV, Reiss JI, Murray PS, Dishman RK, Spradley JM. Chronic exercise dampens hippocampal glutamate overflow induced by kainic acid in rats. Behavioural Brain Research. 2015;284:19-23. https://doi.org/10.1016/j.bbr.2015.02.002

  • Receive Date 14 July 2025
  • Revise Date 13 September 2025
  • Accept Date 06 October 2025
  • First Publish Date 06 October 2025
  • Publish Date 22 June 2026