Pre-Ischemic Oxytocin Treatment Alleviated Neuronal Injury via Suppressing NF-κB, MMP-9, and Apoptosis Regulator Proteins in A Mice Model of Stroke

Document Type : Original Article


1 Research Center of Physiology, Semnan University of Medical Sciences, Semnan, Iran

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

3 Department of Physiology, Faculty of Medicine, Semnan University of Medical Sciences, Semnan, Iran *Corresponding Address: P.O.Box: 3513138111, Department of Physiology, Faculty of Medicine, Semnan University of Medical


Objective: This study was designed to determine the effects of pre-ischemic administration of oxytocin (OXT) on neuronal injury and possible molecular mechanisms in a mice model of stroke.
Materials and Methods: In this experimental study, stroke was induced in the mice by middle cerebral artery occlusion (MCAO) for 60 minutes and 24 hours of reperfusion. OXT was given as intranasal daily for 7 consecutive days before ischemic stroke. Neuronal damage, spatial memory, and the expression levels of nuclear factor-kappa B (NF-κB), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), matrix metalloproteinase-9 (MMP-9), brain-derived neurotrophic factor (BDNF) and apoptosis were assessed 24 hours after stroke.
Results: Pre-ischemic treatment with OXT significantly reduced the infarct size (P<0.01); but did not recover the neurological and spatial memory dysfunction (P>0.05). Moreover, OXT treatment considerably decreased the expressions of NF-κB, TNF-α, IL-1β, and MMP-9 (P<0.001) and enhanced the level of BDNF protein. OXT treatment also significantly downregulated Bax expression and overexpressed Bcl-2 proteins.
Conclusion: The finding of this study indicated that administration of OXT before ischemia could limit brain injury by inhibiting MMP-9 expression, apoptosis, inflammatory signaling pathways, and an increase in the BDNF protein level. We suggested that OXT may be potentially useful in the prevention and/or reducing the risk of the cerebral stroke attack, and could be offered as a new prevention option in the clinics.


1. Campbell BCV, De Silva DA, Macleod MR, Coutts SB, Schwamm LH, Davis SM, et al. Ischaemic stroke. Nat Rev Dis Primers. 2019; 5(1): 70.
2. Karimipour M, Shojaei Zarghani S, Mohajer Milani M, Soraya H. Pre-treatment with metformin in comparison with post-treatment reduces cerebral ischemia reperfusion induced injuries in rats. Bull Emerg Trauma. 2018; 6(2): 115-121.
3. Famakin BM, Chimowitz MI, Lynn MJ, Stern BJ, George MG. Causes and severity of ischemic stroke in patients with symptomatic intracranial arterial stenosis. Stroke. 2009; 40(6): 1999-2003.
4. Asahi M, Wang X, Mori T, Sumii T, Jung JC, Moskowitz MA, et al. Effects of matrix metalloproteinase-9 gene knock-out on the proteolysis of blood-brain barrier and white matter components after cerebral ischemia. J Neurosci. 2001; 21(19): 7724-7732.
5. Nowacka M, Obuchowicz E. BDNF and VEGF in the pathogenesis of stress-induced affective diseases: an insight from experimental studies. Pharmacol Rep. 2013; 65(3): 535-546.
6. Zhao H, Chen Z, Xie LJ, Liu GF. Suppression of TLR4/NF-κB signaling pathway improves cerebral ischemia-reperfusion injury in rats. Mol Neurobiol. 2018; 55(5): 4311-4319.
7. Prevost M, Zelkowitz P, Tulandi T, Hayton B, Feeley N, Carter CS, et al. Oxytocin in pregnancy and the postpartum: relations to labor and its management. Front Public Health. 2014; 2: 1.
8. Macdonald K, Feifel D. Helping oxytocin deliver: considerations in the development of oxytocin-based therapeutics for brain disorders. Front Neurosci. 2013; 7: 35.
9. Cai Q, Feng L, Yap KZ. Systematic review and meta-analysis of reported adverse events of long-term intranasal oxytocin treatment for autism spectrum disorder. Psychiatry Clin Neurosci. 2018; 72(3): 140-151.
10. Domes G, Ower N, von Dawans B, Spengler FB, Dziobek I, Bohus M, et al. Effects of intranasal oxytocin administration on empathy and approach motivation in women with borderline personality disorder: a randomized controlled trial. Transl Psychiatry. 2019; 9(1): 328.
11. Karelina K, Stuller KA, Jarrett B, Zhang N, Wells J, Norman GJ, et al. Oxytocin mediates social neuroprotection after cerebral ischemia. Stroke. 2011; 42(12): 3606-3611.
12. Etehadi Moghadam S, Azami Tameh A, Vahidinia Z, Atlasi MA, Hassani Bafrani H, Naderian H. Neuroprotective effects of oxytocin hormone after an experimental stroke model and the Possible role of calpain-1. J Stroke Cerebrovasc Dis. 2018; 27(3): 724-732.
13. Momenabadi S, Vafaei AA, Bandegi AR, Zahedi-Khorasani M, Mazaheri Z, Vakili A. Oxytocin reduces brain injury and maintains blood-brain barrier integrity after ischemic stroke in mice. Neuromolecular Med. 2020; 22(4): 557-571.
14. Dayi A, Cetin F, Sisman AR, Aksu I, Tas A, Gönenc S, et al. The effects of oxytocin on cognitive defect caused by chronic restraint stress applied to adolescent rats and on hippocampal VEGF and BDNF levels. Med Sci Monit. 2015; 21: 69-75.
15. Wu Y, Wu T, Xu B, Xu X, Chen H, Li X. Oxytocin prevents cartilage matrix destruction via regulating matrix metalloproteinases. Biochem Biophys Res Commun. 2017; 486(3): 601-606.
16. Behrouzifar S, Vakili A, Bandegi AR, Kokhaei P. Neuroprotective nature of adipokine resistin in the early stages of focal cerebral ischemia in a stroke mouse model. Neurochem Int. 2018; 114: 99- 107.
17. Khorasani MZ, Hosseinzadeh SA, Vakili A. Effect of central microinjection of carbenoxolone in an experimental model of focal cerebral ischemia. Pak J Pharm Sci. 2009; 22(4): 349-354.
18. Akhoundzadeh K, Vakili A, Sameni HR, Vafaei AA, Rashidy-Pour A, Safari M, Mohammadkhani R. Effects of the combined treatment of bone marrow stromal cells with mild exercise and thyroid hormone on brain damage and apoptosis in a mouse focal cerebral ischemia model. Metab Brain Dis. 2017; 32(4): 1267-1277.
19. González-Falcón A, Candelario-Jalil E, García-Cabrera M, León OS. Effects of pyruvate administration on infarct volume and neurological deficits following permanent focal cerebral ischemia in rats. Brain Res. 2003; 990(1-2): 1-7.
20. Stroke Therapy Academic Industry Roundtable II (STAIR-II). Recommendations for clinical trial evaluation of acute stroke therapies. Stroke. 2001; 32(7): 1598-1606.
21. Zhang W, Potrovita I, Tarabin V, Herrmann O, Beer V, Weih F, et al. Neuronal activation of NF-kappaB contributes to cell death in cerebral ischemia. J Cereb Blood Flow Metab. 2005; 25(1): 30-40.
22. Jin R, Liu L, Zhang S, Nanda A, Li G. Role of inflammation and its mediators in acute ischemic stroke. J Cardiovasc Transl Res. 2013; 6(5): 834-851.
23. Ahmed MA, Elosaily GM. Role of oxytocin in deceleration of early atherosclerotic inflammatory processes in adult male rats. Int J Clin Exp Med. 2011; 4(3): 169-178.
24. Khori V, Alizadeh AM, Khalighfard S, Heidarian Y, Khodayari H. Oxytocin effects on the inhibition of the NF-κB/miR195 pathway in mice breast cancer. Peptides. 2018; 107: 54-60.
25. Isenmann S, Stoll G, Schroeter M, Krajewski S, Reed JC, Bähr M. Differential regulation of Bax, Bcl-2, and Bcl-X proteins in focal cortical ischemia in the rat. Brain Pathol. 1998; 8(1): 49-62.
26. Zhang W, Meng A. MicroRNA‑124 expression in the brains of rats during early cerebral ischemia and reperfusion injury is associated with cell apoptosis involving STAT3. Exp Ther Med. 2019; 17(4): 2870-2876.
27. Asadi Y, Gorjipour F, Behrouzifar S, Vakili A. Irisin peptide protects brain against ischemic injury through reducing apoptosis and enhancing BDNF in a rodent model of stroke. Neurochem Res. 2018; 43(8): 1549-1560.
28. Mostafa DG, Khaleel EF, Abdel-Aleem GA. Mechanism of action of oxytocin as cardioprotection in rat model of myocardial infarction. IOSR J Dent Med Sci 2015; 14: 25-36.
29. Al-Amran FF, Shahkolahi M. Oxytocin ameliorates the immediate myocardial injury in heart transplant through down regulation of the neutrophil dependent myocardial apoptosis. Heart Views. 2014; 15(2): 37-45.
30. Latt HM, Matsushita H, Morino M, Koga Y, Michiue H, Nishiki T, et al. Oxytocin inhibits corticosterone-induced apoptosis in primary hippocampal neurons. Neuroscience. 2018; 379: 383-389.
31. Lee SR, Tsuji K, Lee SR, Lo EH. Role of matrix metalloproteinases in delayed neuronal damage after transient global cerebral ischemia. J Neurosci. 2004; 24(3): 671-678.
32. Wang X, Jung J, Asahi M, Chwang W, Russo L, Moskowitz MA, et al. Effects of matrix metalloproteinase-9 gene knock-out on morphological and motor outcomes after traumatic brain injury. J Neurosci. 2000; 20(18): 7037-7042.
33. Gao N, Guo T, Luo H, Tu G, Niu F, Yan M, et al. Association of the MMP-9 polymorphism and ischemic stroke risk in southern Chinese Han population. BMC Neurol. 2019; 19(1): 67.
34. Dang AB, Tay BKB, Kim HT, Nauth A, Alfonso-Jaume MA, Lovett DH. Inhibition of MMP2/MMP9 after spinal cord trauma reduces apoptosis. Spine. 2008; 33(17): E576-E579.
35. Rosell A, Cuadrado E, Ortega-Aznar A, Hernández-Guillamon M, Lo EH, Montaner J. MMP-9-positive neutrophil infiltration is associated to blood-brain barrier breakdown and basal lamina type IV collagen
degradation during hemorrhagic transformation after human ischemic stroke. Stroke. 2008; 39(4): 1121-1126.
36. Turner RJ, Sharp FR. Implications of MMP9 for blood brain barrier disruption and hemorrhagic transformation following ischemic stroke. Front Cell Neurosci. 2016; 10: 56.
37. Rosell A, Ortega-Aznar A, Alvarez-Sabín J, Fernández-Cadenas I, Ribó M, Molina CA, et al. Increased brain expression of matrix metalloproteinase-
9 after ischemic and hemorrhagic human stroke. Stroke. 2006; 37(6): 1399-1406.
38. Könnecke H, Bechmann I. The role of microglia and matrix metalloproteinases involvement in neuroinflammation and gliomas. Clin Dev Immunol. 2013; 2013: 914104.
39. Barker V, Walker RM, Evans KL, Lawrie SM. Methylation of glucocorticoid receptor (NR3C1), BDNF and oxytocin receptor genes in association with childhood maltreatment in schizophrenia and schizoaffective disorder. Schizophr Res. 2020; 216: 529-531.
40. Camerino C, Conte E, Carratù MR, Fonzino A, Lograno MD, Tricarico D. Oxytocin/osteocalcin/IL-6 and NGF/BDNF mRNA levels in response to cold stress challenge in mice: possible oxytonic brainbone- muscle-interaction. Front Physiol. 2019; 10: 1437.