Intracerebroventricular injection of Wharton’s jelly mesenchymal stem cells attenuates Brain Damage in Rat Model of Hypoxia: Optimization of vascular endothelial growth factor and downregulation of inflammatory factors
AbstractObjective: In this study, we investigated the effects of intracerebroventricular (ICV) Wharton’s jelly mesenchymal stem cells (WJ-MSCs) injection in rat model of hypoxic brain injury by evaluating the amount of vascular endothelial growth factor (VEGF) and pro inflammatory factors in hippocampus.
Methods: 24 rats were allocated to four groups of study:1) control and intact animals (Co), 2) sham group (Sh): animals were placed in the hypoxia chamber without inducing hypoxia and injected PBS, 3) hypoxia (H), 4) H+WJ-MSC, Hypoxia was induced by placing animals in the hypoxia chamber for 30 days (4hours a day). After three days of vehicle or WJ-MSCs injection, the rats were sacrificed and brain tissues were prepared for molecular and histopathological studies.
Results: Despite a decrease in the gene expression of IL1β, TNFα, IL18, and the number of dark neurons in CA1 region of hippocampus in H+ WJ-MSC groups Compared to H (P˂0.05). There is an increase in all these factors in both H and H+WJ-MSC groups compared to CO and Sh groups (P˂0.05). The gene expression and protein concentration of VEGF increased in both H and H+WJ-MSC groups compared to Co and Sh groups (P˂0.05).
Conclusion: Based on the findings, WJ-MSCs could reduce the number of dark neurons in hippocampus by increasing the VEGF synthesis and reducing inflammation in hypoxic condition.
2. He Y, Yao Y, Tsirka SE, Cao Y. Cell-culture models of the blood–brain barrier. Stroke. 2014;45(8):2514-26.
3. Engelhardt S, Al‐Ahmad AJ, Gassmann M, Ogunshola OO. Hypoxia selectively disrupts brain microvascular endothelial tight junction complexes through a hypoxia‐inducible factor‐1 (HIF‐1) dependent mechanism. Journal of cellular physiology. 2014;229(8):1096-105.
4. Hassanzadeh G, Quchani SH, Sahraian MA, Abolhassani F, Gilani MAS, Tarzjani MD, et al. Leukocyte Gene Expression and Plasma Concentration in Multiple Sclerosis: Alteration of Transforming Growth Factor-βs, Claudin-11, and Matrix Metalloproteinase-2. Cellular and molecular neurobiology. 2016;36(6):865-72.
5. Zlokovic BV. The blood-brain barrier in health and chronic neurodegenerative disorders. Neuron. 2008;57(2):178-201.
6. Schoch HJ, Fischer S, Marti HH. Hypoxia‐induced vascular endothelial growth factor expression causes vascular leakage in the brain. Brain. 2002;125(11):2549-57.
7. Tang YL, Tang Y, Zhang YC, Qian K, Shen L, Phillips MI. Improved graft mesenchymal stem cell survival in ischemic heart with a hypoxia-regulated heme oxygenase-1 vector. Journal of the American College of Cardiology. 2005;46(7):1339-50.
8. Roy CS, Sherrington CS. On the regulation of the blood‐supply of the brain. The Journal of physiology. 1890;11(1-2):85-158.
9. Ahmad AA, Taboada CB, Gassmann M, Ogunshola OO. Astrocytes and Pericytes Differentially Modulate Blood—Brain Barrier Characteristics during Development and Hypoxic Insult. Journal of Cerebral Blood Flow & Metabolism. 2011;31(2):693-705.
10. Nakagawa S, Deli MA, Kawaguchi H, Shimizudani T, Shimono T, Kittel A, et al. A new blood–brain barrier model using primary rat brain endothelial cells, pericytes and astrocytes. Neurochemistry international. 2009;54(3-4):253-63.
11. Sorby-Adams AJ, Marcoionni AM, Dempsey ER, Woenig JA, Turner RJ. The role of neurogenic inflammation in blood-brain barrier disruption and development of cerebral oedema following acute central nervous system (CNS) Injury. International journal of molecular sciences. 2017;18(8):1788.
12. Bazrafkan M, Nikmehr B, Shahverdi A, Hosseini SR, Hassani F, Poorhassan M, et al. Lipid peroxidation and its role in the expression of NLRP1a and NLRP3 genes in testicular tissue of male rats: A model of spinal cord injury. Iranian biomedical journal. 2018;22(3):151.
13. Nikmehr B, Bazrafkan M, Hassanzadeh G, Shahverdi A, Gilani MAS, Kiani S, et al. The correlation of gene expression of inflammasome indicators and impaired fertility in rat model of spinal cord injury: A time course study. Urology journal. 2017;14(6):5057-63.
14. Mahakizadeh S, Akbari M, Sharifzadeh M, Rastegar T, Abolhassani F, Hassanzadeh G. Wharton’jelly mesenchymal stem cells and insulin effect on BDNF expression in CA1 and CA3 regions of rats’ hippocampus after chronic hypoxia. Journal of Contemporary Medical Sciences. 2018;4(2):63-9.
15. Trounson A, McDonald C. Stem cell therapies in clinical trials: progress and challenges. Cell stem cell. 2015;17(1):11-22.
16. Bang OY. Clinical trials of adult stem cell therapy in patients with ischemic stroke. Journal of Clinical Neurology. 2016;12(1):14-20.
17. Zhu W, Chen J, Cong X, Hu S, Chen X. Hypoxia and serum deprivation‐induced apoptosis in mesenchymal stem cells. Stem cells. 2006;24(2):416-25.
18. Alizamir T, Akbari M, Mokhtari T, Hassanzadeh G. Associated functional motor recovery induced by Intracerebroventricular (ICV) microinjection of Wharton’s jelly mesenchymal stem cells following brain ischemia/reperfusion injury in rat: Decreased dark neurons and Bax gene expression in the cerebral corte. Journal of Contemporary Medical Sciences. 2017;3(12).
19. Huang NF, Li S. Mesenchymal stem cells for vascular regeneration. 2008.
20. Ping Z, Liu W, Kang Z, Cai J, Wang Q, Cheng N, et al. Sulforaphane protects brains against hypoxic–ischemic injury through induction of Nrf2-dependent phase 2 enzyme. Brain research. 2010;1343:178-85.
21. Krysko DV, Berghe TV, D’Herde K, Vandenabeele P. Apoptosis and necrosis: detection, discrimination and phagocytosis. Methods. 2008;44(3):205-21.
22. Back SA. Cerebral white and gray matter injury in newborns: new insights into pathophysiology and management. Clinics in perinatology. 2014;41(1):1-24.
23. Logitharajah P, Rutherford MA, Cowan FM. Hypoxic-ischemic encephalopathy in preterm infants: antecedent factors, brain imaging, and outcome. Pediatric research. 2009;66(2):222-9.
24. Perez A, Ritter S, Brotschi B, Werner H, Caflisch J, Martin E, et al. Long-term neurodevelopmental outcome with hypoxic-ischemic encephalopathy. The Journal of pediatrics. 2013;163(2):454-9. e1.
25. Muttikkal TJE, Wintermark M. MRI patterns of global hypoxic-ischemic injury in adults. Journal of Neuroradiology. 2013;40(3):164-71.
26. Maurya V, Ravikumar R, Bhatia M, Rai R. Hypoxic–Ischemic brain injury in an adult: Magnetic Resonance Imaging findings. medical journal armed forces india. 2016;72(1):75-7.
27. Mokhtari T, Akbari M, Malek F, Kashani IR, Rastegar T, Noorbakhsh F, et al. Improvement of memory and learning by intracerebroventricular microinjection of T3 in rat model of ischemic brain stroke mediated by upregulation of BDNF and GDNF in CA1 hippocampal region. DARU Journal of Pharmaceutical Sciences. 2017;25(1):4.
28. Hedtjärn M, Leverin A-L, Eriksson K, Blomgren K, Mallard C, Hagberg H. Interleukin-18 involvement in hypoxic–ischemic brain injury. Journal of Neuroscience. 2002;22(14):5910-9.
29. Farahabadi A, Akbari M, Pishva AA, Zendedel A, Arabkheradmand A, Beyer C, et al. Effect of progesterone therapy on TNF-α and iNOS gene expression in spinal cord injury model. Acta medica Iranica. 2016;54(6):345-51.
30. Lievre V, Becuwe P, Bianchi A, Bossenmeyer-Pourie C, Koziel V, Franck P, et al. Intracellular generation of free radicals and modifications of detoxifying enzymes in cultured neurons from the developing rat forebrain in response to transient hypoxia. Neuroscience. 2001;105(2):287-97.
31. Perego C, Fumagalli S, De Simoni M-G. Temporal pattern of expression and colocalization of microglia/macrophage phenotype markers following brain ischemic injury in mice. Journal of neuroinflammation. 2011;8(1):174.
32. Garcia JH, Liu K-F, Yoshida Y, Chen S, Lian J. Brain microvessels: factors altering their patency after the occlusion of a middle cerebral artery (Wistar rat). The American journal of pathology. 1994;145(3):728.
33. Liu F, Mccullough LD. Inflammatory responses in hypoxic ischemic encephalopathy. Acta Pharmacologica Sinica. 2013;34(9):1121.
34. Han HS, Yenari MA. Cellular targets of brain inflammation in stroke. Current opinion in investigational drugs (London, England: 2000). 2003;4(5):522-9.
35. Brait VH, Arumugam TV, Drummond GR, Sobey CG. Importance of T lymphocytes in brain injury, immunodeficiency, and recovery after cerebral ischemia. Journal of Cerebral Blood Flow & Metabolism. 2012;32(4):598-611.
36. Lennmyr F, Ata KA, Funa K, Olsson Y, Terént A. Expression of vascular endothelial growth factor (VEGF) and its receptors (Flt-1 and Flk-1) following permanent and transient occlusion of the middle cerebral artery in the rat. Journal of Neuropathology & Experimental Neurology. 1998;57(9):874-82.
37. Zhang ZG, Zhang L, Jiang Q, Zhang R, Davies K, Powers C, et al. VEGF enhances angiogenesis and promotes blood-brain barrier leakage in the ischemic brain. Journal of Clinical Investigation. 2000;106(7):829-38.
38. Yeh W-L, Lu D-Y, Lin C-J, Liou H-C, Fu W-M. Inhibition of hypoxia-induced increase of blood-brain barrier permeability by YC-1 through the antagonism of HIF-1α accumulation and VEGF expression. Molecular pharmacology. 2007.
39. Sun Y, Jin K, Xie L, Childs J, Mao XO, Logvinova A, et al. VEGF-induced neuroprotection, neurogenesis, and angiogenesis after focal cerebral ischemia. The Journal of clinical investigation. 2003;111(12):1843-51.
40. Fernández-López D, Faustino J, Derugin N, Vexler ZS. Acute and chronic vascular responses to experimental focal arterial stroke in the neonate rat. Translational stroke research. 2013;4(2):179-88.
41. Cobbs CS, Chen J, Greenberg DA, Graham SH. Vascular endothelial growth factor expression in transient focal cerebral ischemia in the rat. Neuroscience letters. 1998;249(2-3):79-82.
42. Haqqani AS, Nesic M, Preston E, Baumann E, Kelly J, Stanimirovic D. Characterization of vascular protein expression patterns in cerebral ischemia/reperfusion using laser capture microdissection and ICAT-nanoLC-MS/MS. The FASEB Journal. 2005;19(13):1809-21.
43. Uccelli A, Moretta L, Pistoia V. Mesenchymal stem cells in health and disease. Nature reviews immunology. 2008;8(9):726.
44. Munoz JR, Stoutenger BR, Robinson AP, Spees JL, Prockop DJ. Human stem/progenitor cells from bone marrow promote neurogenesis of endogenous neural stem cells in the hippocampus of mice. Proceedings of the National Academy of Sciences. 2005;102(50):18171-6.
45. Nam HS, Kwon I, Lee BH, Kim H, Kim J, An S, et al. Effects of mesenchymal stem cell treatment on the expression of matrix metalloproteinases and angiogenesis during ischemic stroke recovery. PLoS One. 2015;10(12):e0144218.
46. Quittet M-S, Touzani O, Sindji L, Cayon J, Fillesoye F, Toutain J, et al. Effects of mesenchymal stem cell therapy, in association with pharmacologically active microcarriers releasing VEGF, in an ischaemic stroke model in the rat. Acta biomaterialia. 2015;15:77-88.
47. Zhou L, Lin Q, Wang P, Yao L, Leong K, Tan Z, et al. Enhanced neuroprotective efficacy of bone marrow mesenchymal stem cells co-overexpressing BDNF and VEGF in a rat model of cardiac arrest-induced global cerebral ischemia. Cell death & disease. 2017;8(5):e2774.
48. Shi Y, Su J, Roberts AI, Shou P, Rabson AB, Ren G. How mesenchymal stem cells interact with tissue immune responses. Trends in immunology. 2012;33(3):136-43.
49. Vendrame M, Gemma C, Mesquita DD, Collier L, Bickford PC, Sanberg CD, et al. Anti-inflammatory effects of human cord blood cells in a rat model of stroke. Stem cells and development. 2005;14(5):595-604.