Post-Conditioning Endurance Training Effect on Apoptosis Inhibition of Hippocampus Neurons in Common Carotid Artery Ischemia Models

Abstract

Purpose

Stroke is one of the debilitating diseases that can lead to organ paralysis and reduced productivity. As a non-pharmalogical approach to treatment, endurance training can improve the symptoms of disability and induce repair of damaged tissues. Apoptosis of sensory-motor neurons is one of the most important mechanisms of nerve damage in stroke, and its inhibition in the early hours after ischemia would be very effective in reducing the complications of the disease. We aimed to evaluate if early exercise training following common carotid artery (CCA) ischemia can modulate apoptosis-related markers, influence neuronal survival in the CA1 region of the hippocampus, and improve motor function recovery.

Methods

Thirty-six Wistar male rats were divided in three groups (n=12); control, CCA ischemia, and CCA ischemia+exercise. The ischemic model was created by a 45-min occlusion of the CCA. The Ladder rung test was executed in 2 h, 24 h, 1 week and 8 weeks after ischemia induction to assess motor function capacity. The Bax and BCL2 gene and protein expression levels were quantified with quantitative PCR and histological tissue staining, respectively.

Results

Compared to control, the Bax expression was higher in ischemic rats and was decreased in the exercise group. The Bcl2 expression was lower in ischemic rats and increased after exercise. The walking ability improved after exercise and the dead cells percentage was lower than in no-intervention ischemic group.

Conclusion

Endurance training can reduce the levels of apoptotic factors and improve the motor ability of ischemic rats after 8 weeks of exercise. Thus, it would be an important approach for improving the motor’s postoperative complication.

Keywords

Endurance training / BAX / BCL2 / Apoptosis / Ischemia

References

[1] Aboutaleb N, Shamsaei N, Khaksari M, Erfani S, Rajabi H, Nikbakht F. Pre-ischemic exercise reduces apoptosis in hippocampal CA3 cells after cerebral ischemia by modulation of the Bax/Bcl-2 proteins ratio and prevention of caspase-3 activation. J Physiol Sci, 2015, 65(5): 435-443

[2] AVERT Trial Collaboration group. Efficacy and safety of very early mobilisation within 24 h of stroke onset (avert): a randomised controlled trial. Lancet. 2015;386(9988):46–55. https://doi.org/10.1016/s0140-6736(15)60690-0.

[3] Bedford TG, Tipton CM, Wilson NC, Oppliger RA, Gisolfi CV. Maximum oxygen consumption of rats and its changes with various experimental procedures. J Appl Physiol, 1979, 47(6): 1278-83

[4] Birnbaum Y, Hale SL, Kloner RA. Differences in reperfusion length following 30 minutes of ischemia in the rabbit influence infarct size, as measured by triphenyltetrazolium chloride staining. J Mol Cell Cardiol, 1997, 29(2): 657-66

[5] Cao LM, Dong ZQ, Li Q, Chen X. Treadmill training improves neurological deficits and suppresses neuronal apoptosis in cerebral ischemic stroke rats. Neural Regen Res, 2019, 14(8): 1387-1393

[6] Carmichael ST. Rodent models of focal stroke: size, mechanism, and purpose. NeuroRx, 2005, 2(3): 396-409

[7] Chen CC, Chang CP, Yang CL. An adaptive fall-free rehabilitation mechanism for ischemic stroke rat patients. Sci Rep, 2019, 9(1): 984

[8] Chen X, Zhang X, Xue L, Hao C, Liao W, Wan Q. Treatment with enriched environment reduces neuronal apoptosis in the periinfarct cortex after cerebral ischemia/reperfusion injury. Cell Physiol Biochem, 2017, 4141445-1456

[9] Dehqanizadeh B, Mohammadi ZF, Kalani AHT, Mirghani SJ. Effect of early exercise on inflammatory parameters and apoptosis in CA1 area of the hippocampus following cerebral ischemia-reperfusion in rats. Brain Res Bull, 2022, 182: 102-110

[10] Eslami Z, Mohammadi Z, Sharifian S, Rezaei Ghomi M, Mousavi SV, Farhadi M, Robati NS, Faghfoori Z, Mirghani SJ. Protective effect of aerobic exercise and adenosine on changes in inflammation mediators after transient ischemia of common carotid arteries in male Wistar rats [Original Articles]. J Gorgan Univ Med Sci, 2021, 23(3): 32-9

[11] Eslami Z, Rezaei Ghomi M, Saidi A, Mousavi SV, Farhadi M, Sheikh Robati N, Moghanlou AE, Metz G, Sharifian S, Mirghani SJ. Endurance training and exogenous adenosine infusion mitigate hippocampal inflammation and cell death in a rat model of cerebral ischemia/reperfusion injury: a randomized controlled trial. Arch Neurosci, 2022, 9(1 e119236

[12] Feigin VL, Nichols E, Alam T, Bannick MS, Beghi E, Blake N, Vos T. Global, regional, and national burden of neurological disorders, 1990–2016: a systematic analysis for the Global Burden of Disease Study 2016. Lancet Neurol, 2019, 185): 459-80

[13] Gao X, Xu X, Pang J, Zhang C, Ding JM, Peng X, Cao JM. receptor activation induces mitochondrial dysfunction, oxidative stress and apoptosis in cultured neonatal rat cardiomyocytes. Physiol Res, 2007, 56(5): 559-569

[14] Hoffman-Goetz L, Spagnuolo PA. Effect of repeated exercise stress on caspase 3, Bcl-2, HSP 70 and CuZn-SOD protein expression in mouse intestinal lymphocytes. J Neuroimmunol, 2007, 187(1–2): 94-101

[15] Hofman FM, Taylor CR. Immunohistochemistry. Curr Protoc Immunol, 2013, 103: 21.24.21-21.24.26

[16] Hugues N, Pellegrino C, Rivera C, Berton E, Pin-Barre C, Laurin J. Is high-intensity interval training suitable to promote neuroplasticity and cognitive functions after stroke?. Int J Mol Sci, 2021, 22(6): 3003

[17] Jafari A, Pourrazi H, Nikookheslat S, Baradaran B. Effect of exercise training on Bcl-2 and Bax gene expression in the rat heart. Gene Cell Tissue, 2015, 24 e60174

[18] Kalogeris T, Baines CP, Krenz M, Korthuis RJ. Cell biology of ischemia/reperfusion injury. Int Rev Cell Mol Biol, 2012, 298: 229-317

[19] Kerr AL, Swain RA. Rapid cellular genesis and apoptosis: effects of exercise in the adult rat. Behav Neurosci, 2011, 125(1): 1-9

[20] Kiernan JA. Histological and histochemical methods: theory and practice. 3rd ed. London: Butterworths; 1999.

[21] Kiss Bimbova K, Bacova M, Kisucka A, Galik J, Zavacky P, Lukacova N. Activation of three major signaling pathways after endurance training and spinal cord injury. Mol Neurobiol, 2022, 59(2): 950-967

[22] Kuriakose D, Xiao Z. Pathophysiology and treatment of stroke: present status and future perspectives. Int J Mol Sci, 2020, 21(20): 7609

[23] Lee R, Kermani P, Teng KK, Hempstead BL. Regulation of cell survival by secreted proneurotrophins. Science, 2001, 29455481945-1948

[24] Li F, Geng X, Khan H, Pendy JTJr., Peng C, Li X, Rafols JA, Ding Y. Exacerbation of brain injury by post-stroke exercise is contingent upon exercise initiation timing. Front Cell Neurosci, 2017, 11 311

[25] Li M, Peng J, Wang MD, Song YL, Mei YW, Fang Y. Passive movement improves the learning and memory function of rats with cerebral infarction by inhibiting neuron cell apoptosis. Mol Neurobiol, 2014, 49(1): 216-221

[26] Li C, Xu X, Wang Z, Wang Y, Luo L, Cheng J, Chen SF, Liu H, Wan Q, Wang Q. Exercise ameliorates post-stroke depression by inhibiting PTEN elevation-mediated upregulation of TLR4/NF-κB/NLRP3 signaling in mice. Brain Res, 2020, 1736 146777

[27] Luo L, Li C, Deng Y, Wang Y, Meng P, Wang Q. High-intensity interval training on neuroplasticity, balance between brain-derived neurotrophic factor and precursor brain-derived neurotrophic factor in poststroke depression rats. J Stroke Cerebrovasc Dis, 2019, 28(3): 672-82

[28] Mahalakshmi B, Maurya N, Lee SD, Bharath Kumar V. Possible neuroprotective mechanisms of physical exercise in neurodegeneration. Int J Mol Sci, 2020, 21(16): 5895

[29] Małczyńska-Sims P, Chalimoniuk M, Sułek A. The effect of endurance training on brain-derived neurotrophic factor and inflammatory markers in healthy people and Parkinson’s disease. A narrative review. Front Physiol, 2020, 11 578981

[30] Malek LA, Malek AK, Leszczynski J, Toutounchi S, Elwertowski M, Spiewak M, Domagala P. Carotid clamping time as a risk factor for early restenosis after carotid endarterectomy. Eur J Vasc Endovasc Surg, 2005, 302143-6

[31] Matsuda F, Sakakima H, Yoshida Y. The effects of early exercise on brain damage and recovery after focal cerebral infarction in rats. Acta Physiol (Oxf), 2011, 201(2): 275-287

[32] Metz GA, Merkler D, Dietz V, Schwab ME, Fouad K. Efficient testing of motor function in spinal cord injured rats. Brain Res, 2000, 883(2): 165-77

[33] Metz GA, Whishaw IQ. The ladder rung walking task: a scoring system and its practical application. J Vis Exp. 2009;(28):1204. https://doi.org/10.3791/1204.

[34] Mohri M, Tomoike H, Noma M, Inoue T, Hisano K, Nakamura M. Duration of ischemia is vital for collateral development: repeated brief coronary artery occlusions in conscious dogs. Circ Res, 1989, 64(2): 287-96

[35] Paxinos G, Franklin KBJ. Paxinos and Franklin’s the mouse brain in stereotaxic coordinates. 5th ed. New York; Acdemic Press; 2019.

[36] Radak D, Katsiki N, Resanovic I, Jovanovic A, Sudar-Milovanovic E, Zafirovic S, Mousad S, Isenovic E. Apoptosis and acute brain ischemia in ischemic stroke. Curr Vasc Pharmacol, 2017, 15(2): 115-22

[37] Sarhadi S, Shahidi F, Keshavarzi Z. The effect of two types of training programs on the expression of TrkB, Bax and Bcl-2 genes in male rats after induction of cerebral ischemia. Gene Cell Tissue, 2020, 7(2 e99467

[38] Schmidt-Kastner R, Wietasch K, Weigel H, Eysel UT. Immunohistochemical staining for glial fibrillary acidic protein (GFAP) after deafferentation or ischemic infarction in rat visual system: features of reactive and damaged astrocytes. Int J Dev Neurosci, 1993, 112): 157-74

[39] Seeger JP, Lenting CJ, Schreuder TH, Landman TR, Cable NT, Hopman MT, Thijssen DH. Interval exercise, but not endurance exercise, prevents endothelial ischemia-reperfusion injury in healthy subjects. Am J Physiol Heart Circ Physiol, 2015, 308(4): H351-7

[40] Tadi P, Liu F, Budd LA. Acute stroke. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2025.

[41] Terashi T, Otsuka S, Takada S, Nakanishi K, Ueda K, Sumizono M, Kikuchi K, Sakakima H. Neuroprotective effects of different frequency preconditioning exercise on neuronal apoptosis after focal brain ischemia in rats. Neurol Res, 2019, 41(6): 510-8

[42] Uluç K, Miranpuri A, Kujoth GC, Aktüre E, Başkaya MK. Focal cerebral ischemia model by endovascular suture occlusion of the middle cerebral artery in the rat. J Vis Exp. 2011;(48):1978. https://doi.org/10.3791/1978.

[43] Wang Y, Zhang H, Chai F, Liu X, Berk M. The effects of escitalopram on myocardial apoptosis and the expression of Bax and Bcl-2 during myocardial ischemia/reperfusion in a model of rats with depression. BMC Psychiatry, 2014, 14: 349

[44] Zhang P, Zhang Y, Zhang J, Wu Y, Jia J, Wu J, Hu Y. Early exercise protects against cerebral ischemic injury through inhibiting neuron apoptosis in cortex in rats. Int J Mol Sci, 2013, 14(3): 6074-89

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Beijing Sport University