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

1 Physical Education and Sport Sciences of University of Tehran

2 PhD Student in Exercise Physiology, Aras International Campus, University of Tehran, Tabriz, Iran

3 Professor, Department of Exercise Physiology, Faculty of Sport Sciences and Health, University of Tehran, Tehran, Iran

4 Assistant Professor, Department of Exercise Physiology, Faculty of Sport Sciences and Health, University of Tehran, Tehran, Iran

Abstract

Introduction: Epilepsy is a chronic neurological disorder and the fourth most common neurological disorder in the world, in which a person experiences two or more seizures without a known cause during their lifetime. Inflammation is one of the key factors involved in the exacerbation of epilepsy, increased neuronal excitability, and the onset of seizures. The aim of this study was to investigate the effect of eight weeks of moderate-intensity aerobic training on the levels of MRP8, TNF-α, and IRAK4 proteins in the hippocampus of epileptic rats.

Methods: In this study, 32 male Wistar rats (4–6 weeks old) an average weight of 226.62±14.96 were divided into four groups: control(n=8), sham(n=8), epilepsy(n=8), and epilepsy + moderate-intensity aerobic training(n=8). After one week of the animals' adaptation to the laboratory environment, epilepsy induction was performed by intrahippocampal injection of kainic acid at a dose of one microgram using the stereotaxic method. To assess the success of epilepsy induction, the Racine Behavioral Scale was used. According to the Raisin scale, stage one was staring, stage two was automatic facial movements such as shaking (wet dog tremor), sniffing (nose movements), salivation, chewing, and head movements, stage three was all events of stage two with forelimb clonus and tremors, stage four was repeated tremors and forelimb clonus and falling, stage five was tonic-clonic seizures with running.
The exercise group performed moderate-intensity aerobic training for 40 minutes per session, five days a week, for 8 weeks. At the end of the training period, 48 hours after the last session, the rats were anesthetized using a combination of ketamine and xylazine, and their hippocampal tissues were extracted. Protein levels of MRP8, TNF-α, and IRAK4 were measured using the Western blot method. To examine the differences in MRP-8, IRAK-4, and TNF-α levels between groups, a one-way ANOVA parametric test was used, followed by a Tukey post hoc test, and to examine the differences in the number of seizures, an independent t-test was used at a significance level of 0.05. All calculations were performed using SPSS software Version 21.

Results: Moderate intensity Aerobic training significantly reduced the hippocampal levels of IRAK4 (P<0.0001, F=89/77), MRP8 (P<0.0001, F=184/00), and TNF-α (P<0.0001, F=60/57) in epileptic rats. Aerobic exercise also significantly reduced the number of seizures in epileptic rats (P=0.01, t=2.80).

Conclusion: Regular aerobic exercise can effectively reduce neuroinflammation and improve epileptic conditions by lowering the hippocampal levels of inflammatory markers MRP8, IRAK4, and TNF-α.

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Articles in Press, Accepted Manuscript
Available Online from 06 October 2025
  • Receive Date: 14 July 2025
  • Revise Date: 13 September 2025
  • Accept Date: 06 October 2025
  • First Publish Date: 06 October 2025
  • Publish Date: 06 October 2025