Evaluation of immune response after moderate and overtraining exercise in wistar rat

Document Type : Original Article


1 Neurogenic Inflammation Research Centre and Department of Physiology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Neurocognitive Research Centre, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

3 Immunology Research Center, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran



The effect of prolonged overtraining on cytokine kinetics was compared with moderate exercise in the present study.
Materials and Methods:
Male Wistar rats were randomly divided into control sedentary (C), moderate trained (MT), (V=20 m/min, 30 min/day for 6 days a week, 8 weeks), overtrained (OT) (V=25 m/min, 60min/day for 6 days a week, 11 weeks) and recovered overtrained (OR) (OT plus 2 weeks recovery) groups, (n=6 for each group). Immediately, 24 hr and 2 weeks (in OR) after last bout of exercise blood samples were obtained. The plasma concentrations of TNFα, IL-6, IL-10, IL-4 and IFN were measured by ELISA method.
Immediately after last bout of exercise the following findings were observed; IL-6, IL-10 and TNFαconcentrations increased in OT and OR groups compared with control (P<0.05–P<0.001). Serum level of IL-4 decreased (P<0.01) but IFN increased (P<0.05) in MT group vs. control. In addition, circulatory levels of TNFα IL-6, IL-10 and IL-4 were higher but the IFN concentrations were lower in OT and OR groups than MT group (P<0.05-P<0.01). The IFN-γIL4 ratio was significantly increased in MT (P<0.01) while it decreased in OT group. There were not statistical differences in TNFα IL-6, and IFN levels between different time intervals after exercise in MT, OT and OR groups.
These data confirm a positive effect of moderate exercise on immune function and a decrease in susceptibility to viral infection by inducing Th1 cytokine profile shift. However, prolonged and overtraining exercise causes numerous changes in immunity that possibly reflects physiological stress and immune suppression.


1. Wen CP, Wu X. Stressing harms of physical inactivity to promote exercise. Lancet. 2012; 380:192-193.
2. Lee IM, Shiroma EJ, Lobelo F, Puska P, Blair SN, Katzmarzyk PT. Effect of physical inactivity on major non-communicable diseases worldwide: an analysis of burden of disease and life expectancy. Lancet. 2012; 380:219-229.
3. Woodcock J, Franco OH, Orsini N, Roberts I. Non-vigorous physical activity and all-cause mortality: systematic review and meta-analysis of cohort studies. Int J Epidemiol. 2011; 40:121-138.
4. Fallah Mohammadi M, Hajizadeh Moghaddam A, Mirkarimpur H. The effects of a moderate exercise program on knee osteoarthritis in male wistar rats. Iran J Basic Med Sci. 2013; 16:683-688.
5. Pimlott N. The miracle drug. Can Fam Physician. 2010; 56:407-409.
6. Eshraghi-Jazi F, Alaei H, Azizi-Malekabadi H, Gharavi-Naini M, Pilehvarian A, Ciahmard Z. The effect of red grape juice and exercise, and their combination on parkinson’s disease in rats. Avicenna Journal of Phytomedicine. 2012; 2:90-96.
7. Organization WH. Global Recommendations on Physical Activity for Health; 2010.
8. Janssen I, Ross R. Vigorous intensity physical activity is related to the metabolic syndrome independent of the physical activity dose. Int J Epidemiol. 2012; 41:1132-1140.
9. Physical Activity Guidelines Advisory Committee report. To the Secretary of Health and Human Services. Part A: executive summary. Nutr Rev. 2009; 67:114–120.
10. Hewitt M, Creel A, Estell K, Davis IC, Schwiebert LM. Acute exercise decreases airway inflammation, but not responsiveness, in an allergic asthma model. Am J Respir Cell Mol Biol. 2009; 40:83-89.
11. Gleeson M. Cytokines and exercise. In: Gleeson M, editor. Immune function in sport and exercise. Edinburgh: Elsevier; 2006. p. ix-xi.
12. Kilciler G, Musabak U, Bagci S, Yesilova Z, Tuzun A, Uygun A,
et al. Do the changes in the serum levels of IL-2, IL-4, TNF alpha, and IL-6 reflect the inflammatory activity in the patients with post-ERCP pancreatitis? Clin Dev Immunol. 2008; 2008:481560.
13. Dinarello CA. Interleukin-1, interleukin-1 receptors and interleukin-1 receptor antagonist. Int Rev Immunol. 1998; 16:457-499.
14. Mosmann TR, Cherwinski H, Bond MW, Giedlin MA, Coffman RL. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol. 1986; 136:2348-2357.
15. Abbas AK, Lichtman AH, Pillai S. Immunity to Microbes. Cellular and molecular immunology. Philadelphia: Elsevier/Saunders; 2012; p.345-363.
Gholamnezhad et al Effect of exercise intensities on immune response
Iran J Basic Med Sci, Vol. 17, No. 1, Jan 2014 7
16. Seder RA, Paul WE. Acquisition of lymphokine-producing phenotype by CD4+ T cells. Annu Rev Immunol. 1994; 12:635-673.
17. Kaiko GE, Horvat JC, Beagley KW, Hansbro PM. Immunological decision-making: how does the immune system decide to mount a helper T-cell response? Immunology. 2008; 123:326-338.
18. Radak Z, Chung HY, Koltai E, Taylor AW, Goto S. Exercise, oxidative stress and hormesis. Ageing Res Rev. 2008; 7:34-42.
19. Kazeem A, Olubayo A, Ganiyu A. Plasma Nitric Oxide and Acute Phase Proteins after Moderate and Prolonged xercises. Iran J Basic Med Sci. 2012; 15:602-607.
20. Oishi K, Nishio N, Konishi K, Shimokawa M, Okuda T, Kuriyama T,
et al. Differential effects of physical and psychological stressors on immune functions of rats. Stress. 2003; 6:33-40.
21. Zhao G, Zhou S, Davie A, Su Q. Effects of moderate and high intensity exercise on T1/T2 balance. Exerc Immunol Rev. 2012; 18:98-114.
22. Moreira A, Delgado L, Moreira P, Haahtela T. Does exercise increase the risk of upper respiratory tract infections? Br Med Bull. 2009; 90:111-131.
23. Petibois C, Cazorla G, Poortmans JR, Deleris G. Biochemical aspects of overtraining in endurance sports: the metabolism alteration process syndrome. Sports Med. 2003; 33:83-94.
24. Ogonovszky H, Sasvári M, Dosek A, Berkes I, Kaneko T, Tahara S,
et al. The effects of moderate, strenuous, and overtraining on oxidative stress markers and DNA repair in rat liver. Canadian Journal of Applied Physiology. 2005; 30:186-195.
25. Hohl R, Ferraresso RL, De Oliveira RB, Lucco R, Brenzikofer R, De Macedo DV. Development and characterization of an overtraining animal model. Med Sci Sports Exerc. 2009; 41:1155-1163.
26. Petersen AM, Pedersen BK. The anti-inflammatory effect of exercise. J Appl Physiol. 2005; 98:1154-1162.
27. Wallberg L, Mikael Mattsson C, Enqvist JK, Ekblom B. Plasma IL-6 concentration during ultra-endurance exercise. Eur J Appl Physiol. 2011; 111:1081-1088.
28. Kim H, Shin MS, Kim SS, Lim BV, Kim HB, Kim YP,
et al. Modulation of immune responses by treadmill exercise in Sprague-Dawley rats. J Sports Med Phys Fitness. 2003; 43:99-104.
29. Lira FS, Rosa JC, Pimentel GD, Tarini VA, Arida RM, Faloppa F,
et al. Inflammation and adipose tissue: effects of progressive load training in rats. Lipids Health Dis. 2010; 9:109-115.
30. Halson SL, Jeukendrup AE. Does overtraining exist? An analysis of overreaching and overtraining research. Sports Med. 2004; 34:967-981.
31. Ostrowski K, Rohde T, Zacho M, Asp S, Pedersen BK. Evidence that interleukin-6 is produced in human skeletal muscle during prolonged running. J Physiol. 1998; 508:949-953.
32. Ostrowski K, Hermann C, Bangash A, Schjerling P, Nielsen JN, Pedersen BK. A trauma-like elevation of plasma cytokines in humans in response to treadmill running. J Physiol. 1998; 513:889-894.
33. Gleeson M. Immune function in sport and exercise. J Appl Physiol. 2007; 103:693-699.
34. Bijeh N, Nazem F, Tavakol Afshari J, Nejat Shokouhi A, Mahmoudi M, Rastin M. The effect of eccentric exercise patterns on immune system parameters of female athletes. Iran J Basic Med Sci. 2000; 3:13-19.
35. Pedersen BK, Febbraio MA. Muscle as an endocrine organ: focus on muscle-derived interleukin-6. Physiol Rev. 2008; 88:1379-1406.
36. Febbraio MA, Steensberg A, Keller C, Starkie RL, Nielsen HB, Krustrup P,
et al. Glucose ingestion attenuates interleukin-6 release from contracting skeletal muscle in humans. J Physiol. 2003; 549:607-612.
37. Jonsdottir IH, Schjerling P, Ostrowski K, Asp S, Richter EA, Pedersen BK. Muscle contractions induce interleukin-6 mRNA production in rat skeletal muscles. J Physiol. 2000; 528:157-163.
38. Lancaster GI. Chapter 10 - Exercise and cytokines. In: Gleeson M, editors. Immune Function in Sport and Exercise. Edinburgh: Elsevier; 2006. p. 205-220.
39. Keller C, Steensberg A, Pilegaard H, Osada T, Saltin B, Pedersen BK,
et al. Transcriptional activation of the IL-6 gene in human contracting skeletal muscle: influence of muscle glycogen content. FASEB J. 2001; 15:2748-2750.
40. Keller C, Keller P, Marshal S, Pedersen BK. IL-6 gene expression in human adipose tissue in response to exercise--effect of carbohydrate ingestion. J Physiol. 2003; 550:927-931.
41. Steensberg A, van Hall G, Osada T, Sacchetti M, Saltin B, Klarlund Pedersen B. Production of interleukin-6 in contracting human skeletal muscles can account for the exercise-induced increase in plasma interleukin-6. J Physiol. 2000; 529:237-242.
42. Pedersen BK, Steensberg A, Keller P, Keller C, Fischer C, Hiscock N,
et al. Muscle-derived interleukin-6: lipolytic, anti-inflammatory and immune regulatory effects. Pflugers Arch. 2003; 446:9-16.
43. Febbraio MA, Hiscock N, Sacchetti M, Fischer CP, Pedersen BK. Interleukin-6 is a novel factor mediating glucose homeostasis during skeletal muscle contraction. Diabetes. 2004; 53:1643-1648.
44. Brandt C, Jakobsen AH, Adser H, Olesen J, Iversen N, Kristensen JM,
et al. IL-6 regulates exercise and training-induced adaptations in subcutaneous adipose tissue in mice. Acta Physiol. 2012; 205:224-235.
45. Suzuki K, Yamada M, Kurakake S, Okamura N, Yamaya K, Liu Q,
et al. Circulating cytokines and hormones with immunosuppressive but neutrophil-priming potentials rise after endurance exercise in humans. Eur J Appl Physiol. 2000; 81:281-287.
46. Steensberg A, Keller C, Starkie RL, Osada T, Febbraio MA, Pedersen BK. IL-6 and TNF-alpha expression in, and release from, contracting human skeletal muscle. Am J Physiol Endocrinol Metab. 2002; 283:E1272-1278.
47. Steensberg A, Fischer CP, Keller C, Moller K, Pedersen BK. IL-6 enhances plasma IL-1ra, IL-10, and cortisol in humans. Am J Physiol Endocrinol Metab. 2003; 285:E433-437.
48. Northoff H, Berg A, Weinstock C. Similarities and differences of the immune response to exercise and trauma: the IFN-gamma concept. Can J Physiol Pharmacol. 1998; 76:497-504.
Effect of exercise intensities on immune response Gholamnezhad et al
Iran J Basic Med Sci, Vol. 17, No. 1, Jan 2014 8
49. Lancaster GI, Halson SL, Khan Q, Drysdale P, Wallace F, Jeukendrup AE,
et al. Effects of acute exhaustive exercise and chronic exercise training on type 1 and type 2 T lymphocytes. Exerc Immunol Rev. 2004; 10:91-106.
50. Webster JI, Tonelli L, Sternberg EM. Neuroendocrine regulation of immunity. Annu Rev Immunol. 2002; 20:125-163.
51. McAlees JW, Smith LT, Erbe RS, Jarjoura D, Ponzio NM, Sanders VM. Epigenetic regulation of beta2-adrenergic receptor expression in T(H)1 and T(H)2 cells. Brain Behav Immun. 2011; 25:408-415.
52. Itoh CE, Kizaki T, Hitomi Y, Hanawa T, Kamiya S, Ookawara T,
et al. Down-regulation of beta2-adrenergic receptor expression by exercise training increases IL-12 production by macrophages following LPS stimulation. Biochem Biophys Res Commun. 2004; 322:979-984.
53. Neto JC, Lira FS, de Mello MT, Santos RV. Importance of exercise immunology in health promotion. Amino Acids. 2011; 41:1165-1172