Voluntary exercise improves pulmonary inflammation through NF-κB and Nrf-2 in type 2 diabetic male rats

Document Type : Original Article


1 Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran

2 Department of Physiology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran

3 Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

4 Tuberculosis and Lung Disease Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

5 Cellular and Molecular Research Center, Research Institute for Prevention of Non-Communicable Disease, Qazvin University of Medical Sciences, Qazvin, Iran

6 Department of Pathobiology, Faculty of Veterinary Medicine, Division of Pathology, Urmia University, Urmia, Iran


Objective(s): This study aimed to evaluate the effects of voluntary exercise as an anti-inflammatory intervention on the pulmonary levels of inflammatory cytokines in type 2 diabetic male rats.
Materials and Methods: Twenty-eight male Wistar rats were divided into four groups (n=7), including control (Col), diabetic (Dia), voluntary exercise (Exe), and diabetic with voluntary exercise (Dia+Exe). Diabetes was induced by a high-fat diet (4 weeks) and intraperitoneal injection of streptozotocin (35 mg/kg), and animals did training on the running wheel for 10 weeks as voluntary exercise. Finally, the rats were euthanized and the lung tissues were sampled for the evaluation of the levels of pulmonary interleukin (IL)-10, IL-11, and TNF-α using ELISA, and the protein levels of Nrf-2 and NF-κB using western blotting and tissue histopathological analysis. 
Results: Diabetes reduced the IL-10, IL-11, and Nrf2 levels (P<0.001 to P<0.01) and increased the levels of TNF-α and NF-κB compared to the Col group (P<0.001). Lung tissue levels of IL-10, IL-11, and Nrf2 in the Dia+Exe group enhanced compared to the Dia group (P<0.001 to P<0.05), however; the TNF-α and NF-κB levels decreased (P<0.001). The level of pulmonary Nrf2 in the Dia+Exe group was lower than that of the Exe group while the NF-κB level increased (P<0.001). Moreover, diabetes caused histopathological changes in lung tissue which improved with exercise in the Dia+Exe group. 
Conclusion: These findings showed that voluntary exercise could improve diabetes-induced pulmonary complications by ameliorating inflammatory conditions.


Main Subjects

1. Khateeb J, Fuchs E, Khamaisi M. Diabetes and lung disease: A neglected relationship. Rev Diabet Stud 2018; 15:e1–e10. 
2. Mirrakhimov AE. Chronic obstructive pulmonary disease and glucose metabolism: a bittersweet symphony. Cardiovasc Diabetol 2012; 11:1–26. 
3. Oguntibeju OO. Type 2 diabetes mellitus, oxidative stress, and inflammation: examining the links. Int J Physiol Pathophysiol Pharmacol 2019; 11:45–63. 
4. Farrah TE, Dhillon B, Keane PA, Webb DJ, Dhaun N. The eye, the kidney, and cardiovascular disease: old concepts, better tools, and new horizons. Kidney Int 2020; 98:323–342. 
5. Leyva-López N, Gutierrez-Grijalva E, Ambriz-Perez D, Heredia J. Flavonoids as cytokine modulators: a possible therapy for inflammation-related diseases. Int J Mol Sci 2016; 17:921–936. 
6. Wardyn JD, Ponsford AH, Sanderson CM. Dissecting molecular cross-talk between Nrf2 and NF-κB response pathways. Biochem Soc Trans 2015; 43:621–626. 
7. Bellezza I, Mierla AL, Minelli A. Nrf2 and NF-κB and their concerted modulation in cancer pathogenesis and progression. Cancers (Basel) 2010; 2:483–497. 
8. Ganesh Yerra V, Negi G, Sharma SS, Kumar A. Potential therapeutic effects of the simultaneous targeting of the Nrf2 and NF-κB pathways in diabetic neuropathy. Redox Biol 2013; 1:394–397. 
9. Kalsi A, Singh S, Taneja N, Kukal S, Mani S. Current treatments for type 2 diabetes, their side effects and possible complementary treatments. Int J Pharm Pharm Sci 2015; 7:1–12. 
10. Scheffer D da L, Latini A. Exercise-induced immune system response: Anti-inflammatory status on peripheral and central organs. Biochim Biophys Acta - Mol Basis Dis 2020; 1866:1–15. 
11. Golbidi S, Badran M, Laher I. Anti-oxidant and anti-inflammatory effects of exercise in diabetic patients. Exp Diabetes Res 2012; 2012:1–16. 
12. Nemmar A, Al-Salam S, Yuvaraju P, Beegam S, Ali BH. Exercise training mitigates water pipe smoke exposure-induced pulmonary impairment via inhibiting NF- κ B and activating Nrf2 signalling pathways. Oxid Med Cell Longev 2018; 2018:1–10. 
13. Pedersen BK. Anti-inflammatory effects of exercise: role in diabetes and cardiovascular disease. Eur J Clin Invest 2017; 47:600–611. 
14. Srinivasan K, Viswanad B, Asrat L, Kaul CL, Ramarao P. Combination of the high-fat diet-fed and low-dose streptozotocin-treated rat: a model for type 2 diabetes and pharmacological screening. Pharmacol Res 2005; 52:313–320. 
15. Chodari L, Mohammadi M, Ghorbanzadeh V, Dariushnejad H, Mohaddes G. Testosterone and voluntary exercise promote angiogenesis in hearts of rats with diabetes by enhancing expression of vegf-a and sdf-1a. Can J Diabetes 2016; 40:436–441. 
16. Ren Y, Du C, Shi Y, Wei J, Wu H, Cui H. The Sirt1 activator, SRT1720, attenuates renal fibrosis by inhibiting CTGF and oxidative stress. Int J Mol Med 2017; 39:1317–1324. 
17. Minato K, Miyake Y, Fukumoto S, Yamamoto K, Kato Y, Shimomura Y, et al. Lemon flavonoid, eriocitrin, suppresses exercise-induced oxidative damage in rat liver. Life Sci 2003; 72:1609–1616. 
18. Tsalamandris S, Antonopoulos AS, Oikonomou E, Papamikroulis G-A, Vogiatzi G, Papaioannou S, et al. The role of inflammation in diabetes: current concepts and future perspectives. Eur Cardiol Rev 2019; 14:50–59. 
19. Klein OL, Krishnan JA, Glick S, Smith LJ. Systematic review of the association between lung function and Type 2 diabetes mellitus. Diabet Med 2010; 27:977–987. 
20. Ofulue AF, Thurlbeck WM. Experimental diabetes and the lung. in vivo connective tissue metabolism. Am Rev Respir Dis 1988; 138:284–289. 
21. Popov D, Hasu M, Costache G, Stern D, Simionescu M. Capillary and aortic endothelia interact in situ with nonenzymatically glycated albumin and develop specific alterations in early experimental diabetes. Acta Diabetol 1997; 34:285–293. 
22. Sugahara K, Ushijima K, Morioka T, Usuku G. Studies of the lung in diabetes mellitus. Virchows Arch A Pathol Anat Histol 1981; 390:313–324. 
23. Thyagarajan B, Jacobs DR, Apostol GG, Smith LJ, Lewis CE, Williams OD. Plasma fibrinogen and lung function: the CARDIA Study. Int J Epidemiol 2006; 35:1001–1008. 
24. Hancox RJ, Poulton R, Greene JM, Filsell S, McLachlan CR, Rasmussen F, et al. Systemic inflammation and lung function in young adults. Thorax 2007; 62:1064–1068. 
25. Pradhan AD. C-reactive protein, interleukin 6, and risk of developing type 2 diabetes mellitus. JAMA 2001; 286:327–334. 
26. Gholamnezhad Z, Boskabady MH, Hosseini M, Sankian M, Khajavi Rad A. Evaluation of immune response after moderate and overtraining exercise in wistar rat. Iran J Basic Med Sci 2014; 17:1–8. 
27. Liu T, Zhang L, Joo D, Sun S-C. NF-κB signaling in inflammation. Signal Transduct Target Ther 2017; 2:1–9. 
28. Sheikhpour E, Noorbakhsh P, Foroughi E, Farahnak S, Nasiri R, Neamatzadeh H. A survey on the role of interleukin-10 in breast cancer: A narrative. Reports Biochem Mol Biol 2018; 7:30–37. 
29. Iyer SS, Cheng G. Role of interleukin 10 transcriptional regulation in inflammation and autoimmune disease. Crit Rev Immunol 2012; 32:23–63. 
30. Zhu YP, Brown JR, Sag D, Zhang L, Suttles J. Adenosine 5′-monophosphate–activated protein kinase regulates il-10–mediated anti-inflammatory signaling pathways in macrophages. J Immunol 2015; 194:584–594. 
31. Guragain D, Gurung P, Chang J-H, Katila N, Chang HW, Jeong B-S, et al. AMPK is essential for IL-10 expression and for maintaining the balance between inflammatory and cytoprotective signaling. Biochim Biophys Acta - Gen Subj 2020; 1864:1–11. 
32. Barry JC, Shakibakho S, Durrer C, Simtchouk S, Jawanda KK, Cheung ST, et al. Hyporesponsiveness to the anti-inflammatory action of interleukin-10 in type 2 diabetes. Sci Rep 2016; 6:1–9. 
33. Ploeger HE, Takken T, de Greef MHG, Timmons BW. The effects of acute and chronic exercise on inflammatory markers in children and adults with a chronic inflammatory disease: a systematic review. Exerc Immunol Rev 2009; 15:6–41. 
34. Calegari L, Nunes RB, Mozzaquattro BB, Rossato DD, Dal Lago P. Exercise training improves the IL-10/TNF-α cytokine balance in the gastrocnemius of rats with heart failure. Brazilian J Phys Ther 2018; 22:154–160. 
35. Lakshmi P, Darshan M, Apoorva S, Ashit G, Prasad R, Suchetha A. Interleukin-11 - its role in the vicious cycle of inflammation, periodontitis, and diabetes: a clinicobiochemical cross-sectional study. J Indian Soc Periodontol 2015; 19:159–163. 
36. Lgssiar A, Hassan M, Schott-Ohly P, Friesen N, Nicoletti F, Trepicchio WL, et al. Interleukin-11 inhibits NF-κB and AP-1 activation in islets and prevents diabetes induced with streptozotocin in mice. Exp Biol Med 2004; 229:425–436. 
37. Chan JF-W, Yuan S, Kok K-H, To KK-W, Chu H, Yang J, et al. A familial cluster of pneumonia associated with the 2019 novel coronavirus indicating person-to-person transmission: a study of a family cluster. Lancet 2020; 395:514–523. 
38. Fashi M, Agha-Alinejad H, Asilian Mahabadi H, Rezaei B, Behzad Pakrad B, Rezaei S. The effects of aerobic exercise on NF-κB and TNF-α in lung tissue of male rat. Nov Biomed 2015; 3:131–134. 
39. Arkan MC, Hevener AL, Greten FR, Maeda S, Li Z-W, Long JM, et al. IKK-β links inflammation to obesity-induced insulin resistance. Nat Med 2005; 11:191–198. 
40. Lingappan K. NF-κB in oxidative stress. Curr Opin Toxicol 2018; 7:81–86. 
41. Panahi G, Pasalar P, Zare M, Rizzuto R, Meshkani R. High glucose induces inflammatory responses in HepG2 cells via the oxidative stress-mediated activation of NF-κB, and MAPK pathways in HepG2 cells. Arch Physiol Biochem 2018; 124:468–474. 
42. Shi X, Li D, Deng Q, Li Y, Sun G, Yuan X, et al. NEFAs activate the oxidative stress-mediated NF-κB signaling pathway to induce inflammatory response in calf hepatocytes. J Steroid Biochem Mol Biol 2015; 145:103–112. 
43. Ordonez DG, Lee MK, Feany MB. α-synuclein induces mitochondrial dysfunction through spectrin and the actin cytoskeleton. Neuron 2018; 97:108–124. 
44. Cawthorn WP, Sethi JK. TNF-α and adipocyte biology. FEBS Lett 2008; 582:117–131. 
45. Patel S, Santani D. Role of NF-κB in the pathogenesis of diabetes and its associated complications. Pharmacol Reports 2009; 61:595–603. 
46. Wang M, Zhao J, Zhang H, Li K, Niu L, Wang Y, et al. Potential protective and therapeutic roles of the nrf2 pathway in ocular diseases: An update. Oxid Med Cell Longev 2020; 2020:1–22. 
47. Ahmed SMU, Luo L, Namani A, Wang XJ, Tang X. Nrf2 signaling pathway: Pivotal roles in inflammation. Biochim Biophys Acta - Mol Basis Dis 2017; 1863:585–597. 
48. Montes S, Juárez-Rebollar D, Nava-Ruíz C, Sánchez-García A, Heras-Romero Y, Rios C, et al. Immunohistochemical study of nrf2-anti-oxidant response element as indicator of oxidative stress induced by cadmium in developing rats. Oxid Med Cell Longev 2015; 2015:1–9. 
49. Suryavanshi S V, Kulkarni YA. NF-κβ: a potential target in the management of vascular complications of diabetes. Front Pharmacol 2017; 8:798.
50. Li W, Wu W, Song H, Wang F, Li H, Chen L, et al. Targeting Nrf2 by dihydro-CDDO-trifluoroethyl amide enhances autophagic clearance and viability of β-cells in a setting of oxidative stress. FEBS Lett 2014; 588:2115–2124. 
51. Uruno A, Yagishita Y, Yamamoto M. The Keap1–Nrf2 system and diabetes mellitus. Arch Biochem Biophys 2015; 566:76–84. 
52. Matzinger M, Fischhuber K, Heiss EH. Activation of Nrf2 signaling by natural products-can it alleviates diabetes? Biotechnol Adv 2018; 36:1738–1767. 
53. Tanase DM, Gosav EM, Anton MI, Floria M, Seritean Isac PN, Hurjui LL, et al. Oxidative stress and NRF2/KEAP1/ARE pathway in diabetic kidney disease (DKD): new perspectives. Biomolecules 2022; 12:1227–1252. 
54. Hayes JD, Dinkova-Kostova AT. The Nrf2 regulatory network provides an interface between redox and intermediary metabolism. Trends Biochem Sci 2014; 39:199–218. 
55. Canning P, Sorrell FJ, Bullock AN. Structural basis of Keap1 interactions with Nrf2. Free Radic Biol Med 2015; 88:101–107. 
56. Mallard AR, Spathis JG, Coombes JS. Nuclear factor (erythroid-derived 2)-like 2 (Nrf2) and exercise. Free Radic Biol Med 2020; 160:471–479. 
57. Tsou Y-H, Shih C-T, Ching C-H, Huang J-Y, Jen CJ, Yu L, et al. Treadmill exercise activates the Nrf2 anti-oxidant system to protect the nigrostriatal dopaminergic neurons from MPP+ toxicity. Exp Neurol 2015; 263:50–62. 
58. Oh Y, Jun H-S. Effects of glucagon-like peptide-1 on oxidative stress and Nrf2 signaling. Int J Mol Sci 2017; 19:26–42. 
59. Tu W, Wang H, Li S, Liu Q, Sha H. The anti-inflammatory and anti-oxidant mechanisms of the Keap1/Nrf2/ARE signaling pathway in chronic diseases. Aging Dis 2019; 10:637–651.