Journal of Sports and Biomotor Sciences

Journal of Sports and Biomotor Sciences

Regulation of apoptosis after Spinal Cord Injury: Synergistic Effects of Aerobic Exercise and Resveratrol on BCL2/BAX Balance

Document Type : Original Article

Authors
Hasan Bagheri 1
Ali Yaghoubi 1
Sadegh Cheragh-birjandi 2
1 Department of Sport Sciences, Boj.C., Islamic Azad University, Bojnord, Iran
2 Department of Sport Sciences, Boj.C., Islamic Azad University, Bojnord, Iran.
10.22034/sbs.2025.537467.1147
Abstract
Introduction and Purpose: Spinal Cord Injury (SCI) induces muscle atrophy by altering apoptosis-autophagy pathways. Pharmacological and exercise interventions can mitigate SCI complications. Therefore, this study aimed to investigate the effects of aerobic exercise, resveratrol supplementation, and their combination on the levels of BAX, BCL2, and BCL2/BAX apoptosis pathway proteins in gastrocnemius muscle.
Materials and Methods: Male Wistar rats, aged forty weeks, were categorized into five groups: healthy control, SCI control, SCI supplementation, SCI training, and resveratrol + training SCI. A spinal cord injury was inflicted by releasing a 10-gram weight from a height of 25 mm onto the spinal cord at the T10 segment. The resveratrol supplement was administered intraperitoneally on a daily basis at a dosage of 10 mg/kg. Aerobic exercise was conducted using a weight-support system for 4 weeks, lasting 58 minutes per session, at an intensity of 20 meters per minute. The levels of BAX and BCL2 in the gastrocnemius muscle were quantified using ELISA. The outcomes were examined utilizing an independent- t-test and one-way ANOVA with Tukey's post hoc analysis.
Results: The findings indicated that SCI resulted in a dramatic reduction in BCL2 protein levels and a diminished BCL2/BAX ratio in the gastrocnemius muscle of rats post-SCI, while BAX levels in the gastrocnemius muscle were significantly elevated. Eight weeks of training, combined with resveratrol administration and aerobic exercise, led to a notable elevation in BCL2 protein levels and the BCL2/BAX ratio in the gastrocnemius muscle relative to the SCI control group. Resveratrol administration, both with and without exercise, elevated BCL2 levels and the BCL2/BAX ratio in the gastrocnemius muscle relative to exercise alone. BAX protein levels diminished following all treatments. The findings indicated that the conjunction of exercise and resveratrol administration markedly elevated BCL2 levels and the BCL2/BAX ratio in the gastrocnemius muscle, in contrast to exercise or supplementation alone. In comparison to exercise alone, both supplementation with and without exercise markedly reduced BAX levels.
Discussion and Conclusion: Aerobic exercise, in conjunction with resveratrol administration, appears to suppress apoptosis via influencing the levels of BCL2, BAX, and the muscle BCL2/BAX ratio in rats with spinal cord injury. 



Forty rats were divided into one healthy group and four SCI groups. The SCI groups were subjected to the following interventions: control, aerobic exercise, resveratrol supplementation, and combined aerobic exercise with resveratrol supplementation. Interventions commenced 14 days post-injury induction and lasted for eight weeks. Aerobic exercise training was performed for 58 minutes at a speed of 20 m/min. The rats were supplemented by a 10 mg dose administered via intraperitoneal injection.



The results showed that SCI significantly decreased BCL2 protein levels and the BCL2/BAX ratio, while significantly increasing BAX levels. Furthermore, eight weeks of aerobic exercise and the combined intervention (exercise + resveratrol) significantly increased BCL2 levels and the BCL2/BAX ratio. Resveratrol supplementation, both with and without exercise, elevated BCL2 levels and the BCL2/BAX ratio more significantly than exercise alone. However, BAX protein levels decreased in response to all interventions. The results also indicated that the combined exercise and resveratrol intervention significantly increased BCL2 and the BCL2/BAX ratio compared to either exercise or supplementation alone. Additionally,

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These findings suggest that aerobic exercise, resveratrol supplementation, and their combination can inhibit apoptosis by modulating the levels of key proteins involved in this pathway in rats with spinal cord injury.
Keywords
Aerobic Training
Resveratrol
Apoptosis
Spinal Cord Injury
Autophagy
Subjects

1. Introduction and Purpose

Spinal Cord Injury (SCI) is a progressive neurological condition characterized by immediate mechanical damage (primary injury), followed by a cascade of secondary pathological events including inflammation, oxidative stress, mitochondrial dysfunction, and programmed cell death (apoptosis). These secondary processes exacerbate tissue damage, leading to persistent motor and sensory deficits. Resveratrol (RSV), a natural polyphenol abundant in grapes, berries, and peanuts, has emerged as a potent neuroprotective agent with multifaceted biological activities. Experimental evidence suggests that RSV administration following SCI reduces oxidative stress and mitigates inflammation. Furthermore, post-SCI use of RSV decreases apoptosis by downregulating the expression of BAX and Caspase-3 while simultaneously upregulating BCL2 expression. On the other hand, aerobic exercise provides neuroprotection by improving blood flow, enhancing the expression of neurotrophic factors, and modulating immune responses. In animal models of SCI, aerobic training (such as treadmill running at 20 m/min) reduces pro-inflammatory cytokines and increases anti-inflammatory mediators in the spinal cord tissue. However, the specific impact of exercise on apoptotic regulators like BAX and BCL2 following SCI remains largely unknown. Given the complex pathophysiology of SCI the initial injury followed by secondary damage expansion a definitive treatment for spinal cord repair has yet to be identified. Since the secondary phase of SCI is associated with significant functional decline, apoptosis, and muscle/tissue atrophy, one of the most critical therapeutic goals for SCI is to slow disease progression, inhibit secondary injury, and improve patient function. Therefore, the aim of this study was to investigate the effect of aerobic training combined with Resveratrol supplementation on the levels of BAX, BCL2, and the BCL2/BAX ratio in the gastrocnemius muscle of rats following SCI.

2. Materials and Methods

Forty male Wistar rats, 8 weeks old, were divided into 5 groups: Sham Control, SCI Control, SCI Supplementation (RSV), SCI Exercise, and SCI Resveratrol + Exercise. SCI was induced by dropping a 10 g weight from a height of 25 mm onto the spinal cord at the T10 segment. Resveratrol supplementation was administered daily via intraperitoneal (IP) injection at a dose of 10 mg/kg of body weight. Aerobic training, utilizing a weight-support system, was conducted for 4 weeks, with each session lasting 58 minutes at a speed of 20 m/min. Forty-eight hours after the final training session, rats were anesthetized using a combined IP injection. The right gastrocnemius muscle was then harvested from all rats. After excision and cleaning of blood, the muscle tissue was transferred to micro-tubes, snap-frozen using liquid nitrogen, and subsequently stored at 70 C. The protein levels of BAX and BCL2 in the gastrocnemius muscle were measured using the ELISA method. Data were analyzed using the Independent t-test and One-way Analysis of Variance (ANOVA) followed by the Tukey's post-hoc test.

3. Results

The results indicated that SCI was associated with a significant decrease in the protein level of BCL2 and the BCL2/BAX ratio in the rats’ gastrocnemius muscle post-SCI, while the BAX level in the gastrocnemius muscle showed a significant increase. Four weeks of exercise training and the combined intervention of Resveratrol supplementation with aerobic exercise resulted in a significant increase in gastrocnemius muscle BCL2 protein level and the BCL2/BAX ratio compared to the SCI Control group. Furthermore, Resveratrol supplementation, both with and without exercise, led to increased BCL2 levels and the BCL2/BAX ratio when compared to the Exercise-only group. BAX protein level decreased in response to all interventions. The results also demonstrated that the combination of exercise and Resveratrol supplementation significantly enhanced BCL2 and the BCL2/BAX ratio in the gastrocnemius muscle compared to exercise or supplementation alone. Moreover, compared to exercise alone, supplementation (with and without exercise) resulted in a significant decrease in BAX.

4. Conclusion

Previous research has shown that BAX and BCL2, which act as pro-apoptotic and anti-apoptotic factors, respectively, cooperatively form a heterodimer during acute SCI that determines the pathway of apoptosis progression. High levels of SCI-induced apoptosis can exacerbate neural damage. This study suggests that Resveratrol and aerobic exercise interactively modulate the BCL2/BAX axis to inhibit apoptosis in SCI rats. The superiority of Resveratrol over exercise in reducing BAX and increasing the BCL2/BAX ratio highlights its strong anti-apoptotic pharmacology, while their combination benefits from complementary mechanisms for enhanced efficacy. These findings support integrated pharmacological and rehabilitation approaches to mitigate secondary neuronal degeneration following spinal cord injury. Future studies should investigate translational protocols and long-term functional outcomes to leverage this synergy for the benefit of patients.

5. Acknowledgment & Funding

The authors would like to express their gratitude to all individuals who assisted in conducting this research and achieving its useful and practical results.

6. Ethical Consideration

All animal procedures in this study were conducted in accordance with the guidelines of the institutional and international standards for the care and use of laboratory animals.  All efforts were made to minimize the number of animals used and to reduce suffering, including the use of appropriate anesthesia and humane endpoints.

7. Contribution of authors

All authors have contributed to the article. all authors read and approved the final manuscript.

8. Conflict of interest

The authors declare no conflict of interest.

1. Mohammadi E, Ghaedi K, Esmailie A, Rahgozar S. Gene expression profiling of liver X receptor α and Bcl-2-associated X protein in experimental transection spinal cord-injured rats. The Journal of Spinal Cord Medicine. 2013;36(1):66-71. [10.1179/2045772312Y.0000000032]
2. the regulation of oxidative stress, inflammation and the Bcl‑2 and Bax pathway. Molecular medicine reports. 2015;12(5):7132-8. [10.3892/mmr.2015.4274]
3. Aslan E, Boyacı MG, Güzel H, Pektaş MB. Better neuroprotective profile of caffeic acid phenyl ester over resveratrol in non-traumatic ischemia-reperfusion injury of the spinal cord. British Journal of Neurosurgery. 2024;38(4):889-95. [10.1080/02688697.2021.1999391]
4. Liu C, Shi Z, Fan L, Zhang C, Wang K, Wang BJBr. Resveratrol improves neuron protection and functional recovery in rat model of spinal cord injury. Brain Research. 2011;1374:100-9. [10.1016/j.brainres.2010.11.061]
5. Tang S, Botchway BO, Zhang Y, Wang X, Huang M, Liu X. Resveratrol can improve spinal cord injury by activating Nrf2/HO-1 signaling pathway. Annals of Anatomy-Anatomischer Anzeiger. 2024;251:152180. [10.1016/j.aanat.2023.152180]
6. Zhao H, Mei X, Yang D, Tu G. Resveratrol inhibits inflammation after spinal cord injury via SIRT-1/NF-κB signaling pathway. Neuroscience Letters. 2021;762:136151. [10.1016/j.neulet.2021.136151]
7. Kan S, Liu C, Zhao X, Feng S, Zhu H, Ma B, et al. Resveratrol improves the prognosis of rats after spinal cord injury by inhibiting mitogen-activated protein kinases signaling pathway. Scientific reports. 2023;13(1):19723. [10.1038/s41598-023-46541-x]
8. Zhao H, Chen S, Gao K, Zhou Z, Wang C, Shen Z, et al. Resveratrol protects against spinal cord injury by activating autophagy and inhibiting apoptosis mediated by the SIRT1/AMPK signaling pathway. Neuroscience. 2017;348:241-51. [10.1016/j.neuroscience.2017.02.027]
9. Xiang Z, Zhang S, Yao X, Xu L, Hu J, Yin C, et al. Resveratrol promotes axonal regeneration after spinal cord injury through activating Wnt/β-catenin signaling pathway. Aging (Albany NY). 2021;13(20):23603. [10.18632/aging.203628]
10. Ranaei M, Yaghoubi A. The Effect of Aerobic Training along with Resveratrol Supplementation on Some Inflammatory Index Levels in the Spinal Cord Tissue of Rats after Spinal Cord Injury. Journal of Shahid Sadoughi University of Medical Sciences. 2024. [10.18502/ssu.v32i11.17787]
11. Nouri A, Farzanegi P, Azarbayjani MA. Effects of Resveratrol Supplementation and Exercise on Apoptosis, Lipid Profile, and Expression of Farnesoid X Receptor, Liver X Receptor and Sirtuin 1 Genes in the Liver of Type 1 Diabetic Rats. Medical Laboratory Journal. 2022;16(4):39-46. [10.52547/mlj.16.4.39]
12. Samarghandian S, Pourbagher-Shahri AM, Ashrafizadeh M, Khan H, Forouzanfar F, Aramjoo H, et al. A pivotal role of the nrf2 signaling pathway in spinal cord injury: A prospective therapeutics study. CNS & Neurological Disorders-Drug Targets (Formerly Current Drug Targets-CNS & Neurological Disorders). 2020;19(3):207-19. [10.2174/1871527319666200604175118]
13. Byrnes KR, Fricke ST, Faden AI. Neuropathological differences between rats and mice after spinal cord injury. Journal of Magnetic Resonance Imaging. 2010;32(4):836-46. [10.1002/jmri.22323]
14. Basso D, Beattie M, Bresnahan J, Anderson D, Faden A, Gruner J, et al. MASCIS evaluation of open field locomotor scores: effects of experience and teamwork on reliability. Journal of neurotrauma. 1996;13(7):343-59. [10.1089/neu.1996.13.343]
15. Girbovan C, Kent P, Merali Z, Plamondon H. Dose-related effects of chronic resveratrol administration on neurogenesis, angiogenesis, and corticosterone secretion are associated with improved spatial memory retention following global cerebral ischemia. Nutritional Neuroscience. 2016;19(8):352-68. [10.1179/1476830515Y.0000000020]
16. Alvarez-Mejia L, Morales J, Cruz GJ, Olayo M-G, Olayo R, Díaz-Ruíz A, et al. Functional recovery in spinal cord injured rats using polypyrrole/iodine implants and treadmill training. Journal of Materials Science: Materials in Medicine. 2015;26(7):209. [10.1007/s10856-015-5541-0]
17. Jung SY, Kim DY, Yune TY, Shin DH, Baek SB, Kim CJ. Treadmill exercise reduces spinal cord injury‑induced apoptosis by activating the PI3K/Akt pathway in rats. Experimental and therapeutic medicine. 2014;7(3):587-93. [10.3892/etm.2013.1451]
18. Serasinghe MN, Missert DJ, Asciolla JJ, Podgrabinska S, Wieder S, Izadmehr S, et al. Anti-apoptotic BCL-2 proteins govern cellular outcome following B-RAFV600E inhibition and can be targeted to reduce resistance. oncogene. 2015;34(7):857-67. [10.1038/onc.2014.21]
19. Kotipatruni RR, Dasari VR, Veeravalli KK, Dinh DH, Fassett D, Rao JS. p53-and Bax-mediated apoptosis in injured rat spinal cord. Neurochemical research. 2011;36:2063-74. [10.1007/s11064-011-0530-2]
20. Sharpe JC, Arnoult D, Youle RJ. Control of mitochondrial permeability by Bcl-2 family members. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research. 2004;1644(2-3):107-13.[10.1016/j.bbamcr.2003.10.016]
21. Hardwick JM, Soane L. Multiple functions of BCL-2 family proteins. Cold Spring Harbor perspectives in biology. 2013;5(2):a008722. [10.1101/cshperspect.a008722]
22. Song W, Kwak H-B, Lawler JMJA. Exercise training attenuates age-induced changes in apoptotic signaling in rat skeletal muscle. Antioxidants & Redox Signaling. 2006;8(3-4):517-28. [10.1089/ars.2006.8.5]
23. Shekarriz H, Galedari M, Nikbakht M, Khorsandi L. Protective effect of continuous aerobic training and crocin on doxorubicin-induced apoptosis in the heart tissue of rats. Jentashapir Journal of Cellular and Molecular Biology. 2022;13(3). [10.5812/jjcmb-130731]
24. Nourzad F, Shahidi F. The effect of aerobic and resistance training on insulin resistance index (HOMA-IR) and BCL-2/BAX ratio in apoptotic pathway in the heart tissue of male wistar diabetic rats. 2022. [10.52547/joeppa.15.1.69] [In Persian]
25. Sedaghat M. Cardiac remodeling, apoptosis-related process (Bax, Bcl-2), and their ratio (Bax/Bcl-2) in cardiomyocytes of diabetic rats after combined exercise training and taurine supplementation. Comparative Clinical Pathology. 2021;30(5):801-10. [10.1007/s00580-021-03275-4]
26. Leonard SS, Xia C, Jiang B-H, Stinefelt B, Klandorf H, Harris GK, et al. Resveratrol scavenges reactive oxygen species and effects radical-induced cellular responses. Biochemical and Biophysical Research Communications. 2003;309(4):1017-26. [10.1016/j.bbrc.2003.08.105]
27. Xu L, Botchway BO, Zhang S, Zhou J, Liu XJFin. Inhibition of NF-κB signaling pathway by resveratrol improves spinal cord injury. Frontiers in neuroscience. 2018;12:690. [10.3389/fnins.2018.00690]
28. Zhang G, Liu Y, Xu L, Sha C, Zhang H, Xu WJBb. Resveratrol alleviates lipopolysaccharide-induced inflammation in PC-12 cells and in rat model. BMC biotechnology. 2019;19:1-9. [10.1186/s12896-019-0502-1]
29. Senturk H, Yaman M, Aydin H, Güney G, Bozkurt I, Paksoy K, et al. Effects of Resveratrol on Inflammation and Apoptosis After Experimental Spinal Cord Injury. Turkish neurosurgery. 2018;28(6). [10.5137/1019-5149.JTN.21829-17.3]
30. Zhao H, Han L, Jian Y, Ma Y, Yan W, Chen X, et al. Resveratrol induces apoptosis in human melanoma cell through negatively regulating Erk/PKM2/Bcl-2 axis. OncoTargets and therapy. 2018:8995-9006. [10.2147/OTT.S186247]
31. Wang P, Jiang L, Zhou N, Zhou H, Liu H, Zhao W, et al. Resveratrol ameliorates autophagic flux to promote functional recovery in rats after spinal cord injury. Oncotarget. 2018;9(9):8427. [10.18632/oncotarget.23877]
32. Graham ZA, Collier L, Peng Y, Saéz JC, Bauman WA, Qin W, et al. A soluble activin receptor IIB fails to prevent muscle atrophy in a mouse model of spinal cord injury. Journal of Neurotrauma. 2016;33(12):1128-35.[ 10.1089/neu.2015.4058]
33. Howard E, Pasiakos S, Blesso C, Fussell M, Rodriguez N. Divergent roles of inflammation in skeletal muscle recovery from injury. Frontier in Physiololgy. 2020; 11: 87. [10. 3389/fphys. 2020.00087]
34. Sun ZK, Ma XR, Jia YJ, Liu YR, Zhang JW, Zhang BA. Effects of resveratrol on apoptosis in a rat model of vascular dementia. Experimental and Therapeutic Medicine. 2014;7(4):843-8. [10.3892/etm.2014.1542]
35. Mahdian H, Farzanegi P, Farzaneh Hesari A. Effect of resveratrol supplementation and interval exercise on heart cells apoptosis in male rats with non-alcoholic fatty liver. Journal of Gorgan University of Medical Sciences. 2020;21(4):39-46. [In Persian]
36. Schrauwen P, Timmers S. Can resveratrol help to maintain metabolic health? Proceedings of the Nutrition Society. 2014;73(2):271-7. [10.1017/S0029665113003856 ]
37. Liao Z-Y, Chen J-L, Xiao M-H, Sun Y, Zhao Y-X, Pu D, et al. The effect of exercise, resveratrol or their combination on Sarcopenia in aged rats via regulation of AMPK/Sirt1 pathway. Experimental Gerontology. 2017;98:177-83. [10.1016/j.exger.2017.08.032]
38. Zhang Z-N, Chung S-K, Xu Z, Xu Y. Oct4 maintains the pluripotency of human embryonic stem cells by inactivating p53 through Sirt1-mediated deacetylation. Stem Cells. 2014;32(1):157-65. [10.1002/stem.1532]

  • Receive Date 07 August 2025
  • Revise Date 19 October 2025
  • Accept Date 25 October 2025