Effects of deferoxamine on the survival of the neuroblastoma SH-SY5Y cells and neuroimmune response in the BV-2 microglial cells
Keywords:
deferoxamine, cell viability, SH-SY5Y, BV-2 cell, IL-10Abstract
Abstract
Microglial are the resident immune cells in the central nervous system (CNS). They release cytokines and chemokines associate with inflammation and consequently lead to the neurological diseases. Iron overload in the CNS is one factor that triggers neuroinflammation. Deferoxamine (DFO) is an iron chelator widely used for removing excessive iron to protect neurons from iron overload. However, a high dose of DFO can induce oxygen depletion and hypoxic damage of neurons. It is still not clear how DFO has an impact on survival and death of cell in the nervous system. Therefore, the objective of this study is to investigate the effects of DFO on cell viability of neuron and microglia cells. The neuroblastoma (SH-SY5Y) and Microglia (BV-2) cell lines were cultured in a completed medium containing DFO at 25, 50, and 100 µM for 24 to 48 hours. Then, the cell viability and the expression of anti-inflammatory cytokine IL-10 were measured. The results showed that after 24 hours of DFO treatment, the cell viability of both cells were significantly decreased as compared to the control. Although the SH-SY5Y cell viability still decreases after 48 hours of DFO treatment, there is a trend to increases of BV-2 cell viability together with a significant increase in the level of IL-10 expression. The finding suggested that DFO treatment, even at low dosage, can induce neuronal and microglial cell death. Furthermore, enhance microglial cell survival and IL-10 expression indicated that the microglia might play an anti-inflammatory role following the hypoxic injury. Our results suggest that DFO should be carefully prescribed to avoid the adverse effects of DFO on hypoxia-induced cell death, especially within 24 hours after drug treatment. Finally, microglia might be a novel therapeutic target for the treatment of neurological diseases related to chronic neuroinflammation.
References
2. Li Y, Pan K, Chen L, et al. Deferoxamine regulates neuroinflammation and iron homeostasis in a mouse model of postoperative cognitive dysfunction. J Neuroinflammation 2016;13:268.
3. Guzman-Martinez L, Maccioni RB, Andrade V, et al. Neuroinflammation as a common feature of neurodegenerative disorders. Front Pharmacol 2019;10:1008.
4. Shao Z, Tu S, Shao A. Pathophysiological mechanisms and potential therapeutic targets in intracerebral hemorrhage. Front Pharmacol 2019;10:1079.
5. Zecca L, Youdim MBH, Riederer P, et al. Iron, brain ageing and neurodegenerative disorders. Nat Rev Neurosci 2004;5:863-73.
6. Belaidi AA, Bush AI. Iron neurochemistry in Alzheimer’s disease and Parkinson’s disease: targets for therapeutics.J Neurochem 2016;139:179-97.
7. Pino JMV, da Luz MHM, Antunes HKM, et al. Iron-restricted diet affects brain ferritin levels, dopamine metabolism and cellular prion protein in a region-specific manner.
Front Mol Neurosci 2017;10:145.
8. Garton T, Keep RF, Hua Y, et al. Brain iron overload following intracranial haemorrhage. Stroke Vasc Neurol 2016;1:172-84.
9. Ward RJ, Zucca FA, Duyn JH, et al. The role of iron in brain ageing and neurodegenerative disorders. Lancet Neurol 2014;13:1045-60.
10. Zeng L, Tan L, Li H, et al. Deferoxamine therapy for intracerebral hemorrhage: A systematic review. PloS one 2018;13:e0193615.
11. Nuñez MT, Chana-Cuevas P. New perspectives in iron chelation therapy for the treatment of neurodegenerative diseases. Pharmaceuticals (Basel)
2018;11:109.
12. Yu Y, Zhao W, Zhu C, et al. The clinical effect of deferoxamine mesylate on edema after intracerebral hemorrhage.PloS one 2015;10:e0122371.
13. Hua Y, Keep R, Hoff J, et al. Deferoxamine therapy for intracerebral hemorrhage. Acta Neurochir Suppl 2008;105:3-6.
14. Wu D, Yotnda P. Induction and Testing of Hypoxia in Cell Culture. J Vis Exp 2011(54):e2899.
15. Milosevic J, Adler I, Manaenko A, et al.Non-hypoxic stabilization of hypoxiainducible factor alpha (HIF-α): relevance in neural progenitor/stem cells. Neurotox Res 2009;15:367-80.
16. Yang Z, Zhao T-z, Zou Y-j, et al. Hypoxia induces autophagic cell death through inducible factor 1α in microglia. PLoS One 2014;9:e96509.
17. Greer SN, Metcalf JL, Wang Y, et al. The updated biology of hypoxia-inducible factor. The EMBO journal 2012;31:2448-60.
18. Chounchay S, Noctor SC, Chutabhakdikul N. Microglia enhances proliferation of neural progenitor cells in an in vitro model of hypoxic-ischemic injury. EXCLI J 2020;19:950-61.
19. Guo M, Song LP, Jiang Y, et al. Hypoxiamimetic agents desferrioxamine and cobalt chloride induce leukemic cell apoptosis through different hypoxia-inducible factor-
1α independent mechanisms. Apoptosis 2006;11:67-77.
20. Kohman RA, Rhodes JS. Neurogenesis, inflammation and behavior. Brain Behav Immun 2013;27:22-32.
21. Kettenmann H, Hanisch U-K, Noda M, et al. Physiology of microglia. Physiol Rev 2011;91:461-553.
22. Lobo-Silva D, Carriche GM, Castro AG, et al. Balancing the immune response in the brain: IL-10 and its regulation. J Neuroinflammation 2016;13:297.
23. Rakshit J, Priyam A, Gowrishetty KK, et al. Iron chelator deferoxamine protects human neuroblastoma cell line SHSY5Y from 6-hydroxydopamine-induced apoptosis and autophagy dysfunction.J Trace Elem Med Biol 2020;57:126406.
24. Woo KJ, Lee T-J, Park J-W, et al. Desferrioxamine, an iron chelator, enhances HIF-1α accumulation via cyclooxygenase-2 signaling pathway. Biochem Biophys Res Commun 2006;343:8-14.
25. Sarah J Texel 1, Jian Zhang, Simonetta Camandola, et al. Ceruloplasmin deficiency reduces levels of iron and BDNF in the cortex and striatum of young mice and increases their vulnerability to stroke. PLoS One 2011;6:e25077.
26. Muñoz P, Humeres A. Iron deficiency on neuronal function. Biometals 2012;25: 825-35.
27. Muñoz P, Humeres A, Elgueta C, et al, Núñez MT. Iron mediates N-methyl-Daspartate receptor-dependent stimulation of calcium-induced pathways and hippocampal synaptic plasticity. J Biol Chem 2011;286:13382-92.
28. He X-f, Lan Y, Zhang Q, et al. Deferoxamine inhibits microglial activation, attenuates blood–brain barrier disruption, rescues dendritic damage, and improves spatial memory in a mouse model of microhemorrhages. J Neurochem 2016;138:436-47.
29. Davis CK, Jain SA, Bae O-N, et al. Hypoxia mimetic agents for ischemic stroke. Front Cell Dev Biol 2019;6:175.
30. Wang X, Ma J, Fu Q, et al. Role of hypoxia-inducible factor-1α in autophagic cell death in microglial cells induced by hypoxia. Mol Med Rep 2017;15:2097-105.
31. Thored P, Heldmann U, Gomes-Leal W, et al. Long-term accumulation of microglia with proneurogenic phenotype concomitant with persistent neurogenesis in adult subventricular zone after stroke.Glia 2009;57:835-49.
32. Li S, Liu W, Wang J, et al. The role of TNF-α, IL-6, IL-10, and GDNF in neuronal apoptosis in neonatal rat with hypoxicischemic encephalopathy. Eur Rev Med Pharmacol Sci 2014;18:905-9.
33. Nizet V, Johnson RS. Interdependence of hypoxic and innate immune responses. Nat Rev Immunol 2009;9:609-17.
34. Whitney NP, Eidem TM, Peng H, et al. Inflammation mediates varying effects in neurogenesis: relevance to the pathogenesis of brain injury and neurodegenerative disorders. J Neurochem 2009;108:1343-59.
35. Fumagalli S, Perego C, Pischiutta F, et al. The ischemic environment drives microglia and macrophage function. Front Neurol 2015;6:81.
