Abstract
Purpose
Intracranial hemorrhage (ICH) is a devastating disease with high mortality and morbidity. After ICH, iron released from the hematoma plays a crucial role in secondary brain injury. Deferasirox (DFR) is a trivalent iron chelator, which was approved to treat iron overload syndrome after transfusion. The aim of the present study was to investigate the protective effects of DFR in both in vitro and in vivo ICH models.
Methods
Using a hemin-induced SH-SY5Y cell damage model, we performed an intracellular bivalent iron (Fe2+) accumulation assay, cell death assay, oxidative stress assessments, and Western blotting analysis. Moreover, the effects of DFR intraventricular administration on hematoma, neurological deficits, and histological alteration were evaluated in an in vivo ICH mouse model by collagenase.
Results
DFR significantly suppressed the intracellular Fe2+ accumulation and cell death caused by hemin exposure. These effects were related to the suppression of both reactive oxygen species and lipid peroxidation over-production. In Western blotting analysis, hemin increased the expression of ferritin (an iron storage protein), LC3 and p62 (autophagy-related markers), phosphorylated p38 (a stress response protein), and cleaved-caspase3 and cleaved-poly (adenosine diphosphate ribose) polymerase (PARP) (apoptosis-related makers). However, DFR suppressed the increase of these proteins. In addition, DFR attenuated the neurological deficits until 7 days after ICH without affecting hematoma and injury area. Furthermore, DFR also suppressed microglia/macrophage activation in peri-hematoma area at 3 days after ICH.
Conclusion
These findings indicate that DFR might be a useful therapeutic agent for the therapy of ICH.
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References
Al-Kuraishy HM, Al-Gareeb AI (2017) Comparison of deferasirox and deferoxamine effects on iron overload and immunological changes in patients with blood transfusion-dependent beta-thalassemia. Asian J Transfus Sci 11:13–17. https://doi.org/10.4103/0973-6247.200768
Arboix A, Comes E, Garcia-Eroles L, Massons J, Oliveres M, Balcells M, Targa C (2002) Site of bleeding and early outcome in primary intracerebral hemorrhage. Acta Neurol Scand 105:282–288. https://doi.org/10.1034/j.1600-0404.2002.1o170.x
Aronowski J, Zhao X (2011) Molecular pathophysiology of cerebral hemorrhage: secondary brain injury. Stroke 42:1781–1786. https://doi.org/10.1161/STROKEAHA.110.596718
Bishop GM, Dang TN, Dringen R, Robinson SR (2011) Accumulation of non-transferrin-bound iron by neurons, astrocytes, and microglia. Neurotox Res 19:443–451. https://doi.org/10.1007/s12640-010-9195-x
Bobinger T, Burkardt P, Huttner BH, Manaenko A (2018) Programmed cell death after intracerebral hemorrhage. Curr Neuropharmacol 16:1267–1281. https://doi.org/10.2174/1570159X15666170602112851
Chelating Agents (2012) LiverTox: Clinical and Research Information on Drug-Induced Liver Injury. Bethesda (MD)
Duan X, Wen Z, Shen H, Shen M, Chen G (2016) Intracerebral hemorrhage, oxidative stress, and antioxidant therapy. Oxidative Med Cell Longev 2016:1203285. https://doi.org/10.1155/2016/1203285
Garton T, Keep RF, Hua Y, Xi G (2016) Brain iron overload following intracranial haemorrhage. Stroke Vasc Neurol 1:172–184. https://doi.org/10.1136/svn-2016-000042
Glickstein H, El RB, Shvartsman M, Cabantchik ZI (2005) Intracellular labile iron pools as direct targets of iron chelators: a fluorescence study of chelator action in living cells. Blood 106:3242–3250. https://doi.org/10.1182/blood-2005-02-0460
Grosso RA, Caldarone PVS, Sanchez MC, Chiabrando GA, Colombo MI, Fader CM (2019) Hemin induces autophagy in a leukemic erythroblast cell line through the LRP1 receptor. Biosci Rep 39. https://doi.org/10.1042/BSR20181156
Gu Y, Hua Y, Keep RF, Morgenstern LB, Xi G (2009) Deferoxamine reduces intracerebral hematoma-induced iron accumulation and neuronal death in piglets. Stroke 40:2241–2243. https://doi.org/10.1161/STROKEAHA.108.539536
Hatakeyama T, Okauchi M, Hua Y, Keep RF, Xi G (2013) Deferoxamine reduces neuronal death and hematoma lysis after intracerebral hemorrhage in aged rats. Transl Stroke Res 4:546–553. https://doi.org/10.1007/s12975-013-0270-5
He Y, Wan S, Hua Y, Keep RF, Xi G (2008) Autophagy after experimental intracerebral hemorrhage. J Cereb Blood Flow Metab: Off J Int Soc Cereb Blood Flow Metab 28:897–905. https://doi.org/10.1038/sj.jcbfm.9600578
Hemorrhagic Stroke Academia Industry Roundtable P (2018) Basic and translational research in Intracerebral hemorrhage: limitations, priorities, and recommendations. Stroke 49:1308–1314. https://doi.org/10.1161/STROKEAHA.117.019539
Hirayama T et al (2017) A universal fluorogenic switch for Fe(ii) ion based on N-oxide chemistry permits the visualization of intracellular redox equilibrium shift towards labile iron in hypoxic tumor cells. Chem Sci 8:4858–4866. https://doi.org/10.1039/c6sc05457a
Ikram MA, Wieberdink RG, Koudstaal PJ (2012) International epidemiology of intracerebral hemorrhage. Curr Atheroscler Rep 14:300–306. https://doi.org/10.1007/s11883-012-0252-1
Imai T, Iwata S, Hirayama T, Nagasawa H, Nakamura S, Shimazawa M, Hara H (2019a) Intracellular Fe(2+) accumulation in endothelial cells and pericytes induces blood-brain barrier dysfunction in secondary brain injury after brain hemorrhage. Sci Rep 9:6228. https://doi.org/10.1038/s41598-019-42370-z
Imai T, Iwata S, Miyo D, Nakamura S, Shimazawa M, Hara H (2019b) A novel free radical scavenger, NSP-116, ameliorated the brain injury in both ischemic and hemorrhagic stroke models. J Pharmacol Sci 141:119–126. https://doi.org/10.1016/j.jphs.2019.09.012
Lan X, Han X, Li Q, Yang QW, Wang J (2017) Modulators of microglial activation and polarization after intracerebral haemorrhage. Nat Rev Neurol 13:420–433. https://doi.org/10.1038/nrneurol.2017.69
Latunde-Dada GO (2017) Ferroptosis: Role of lipid peroxidation, iron and ferritinophagy. Biochim Biophys Acta Gen Subj 1861:1893–1900. https://doi.org/10.1016/j.bbagen.2017.05.019
Li Q, Wan J, Lan X, Han X, Wang Z, Wang J (2017) Neuroprotection of brain-permeable iron chelator VK-28 against intracerebral hemorrhage in mice. J Cereb Blood Flow Metab: Off J Int Soc Cereb Blood Flow Metab 37:3110–3123. https://doi.org/10.1177/0271678X17709186
Quiles Del Rey M, Mancias JD (2019) NCOA4-mediated ferritinophagy: a potential link to neurodegeneration. Front Neurosci 13:238. https://doi.org/10.3389/fnins.2019.00238
Rosenberg GA, Mun-Bryce S, Wesley M, Kornfeld M (1990) Collagenase-induced intracerebral hemorrhage in rats. Stroke 21:801–807. https://doi.org/10.1161/01.str.21.5.801
Ruffels J, Griffin M, Dickenson JM (2004) Activation of ERK1/2, JNK and PKB by hydrogen peroxide in human SH-SY5Y neuroblastoma cells: role of ERK1/2 in H2O2-induced cell death. Eur J Pharmacol 483:163–173. https://doi.org/10.1016/j.ejphar.2003.10.032
Sauerbeck A, Schonberg DL, Laws JL, McTigue DM (2013) Systemic iron chelation results in limited functional and histological recovery after traumatic spinal cord injury in rats. Exp Neurol 248:53–61. https://doi.org/10.1016/j.expneurol.2013.05.011
Selim M et al (2019) Deferoxamine mesylate in patients with intracerebral haemorrhage (i-DEF): a multicentre, randomised, placebo-controlled, double-blind phase 2 trial. Lancet Neurol 18:428–438. https://doi.org/10.1016/S1474-4422(19)30069-9
Sugiyama T, Imai T, Nakamura S, Yamauchi K, Sawada S, Shimazawa M, Hara H (2018) A novel Nrf2 activator, RS9, attenuates secondary brain injury after intracerebral hemorrhage in sub-acute phase. Brain Res 1701:137–145. https://doi.org/10.1016/j.brainres.2018.08.021
Sun YM, Wang YT, Jiang L, Xue MZ (2016) The effects of deferoxamine on inhibition for microglia activation and protection of secondary nerve injury after intracerebral hemorrhage in rats. Pak J Pharm Sci 29:1087–1093
Sung HK, Song E, Jahng JWS, Pantopoulos K, Sweeney G (2019) Iron induces insulin resistance in cardiomyocytes via regulation of oxidative stress. Sci Rep 9:4668. https://doi.org/10.1038/s41598-019-41111-6
Tschoe C, Bushnell CD, Duncan PW, Alexander-Miller MA, Wolfe SQ (2020) Neuroinflammation after Intracerebral Hemorrhage and Potential Therapeutic Targets. J Stroke 22:29–46. https://doi.org/10.5853/jos.2019.02236
Wagner KR, Sharp FR, Ardizzone TD, Lu A, Clark JF (2003) Heme and iron metabolism: role in cerebral hemorrhage. J Cereb Blood Flow Metab: Off J Int Soc Cereb Blood Flow Metab 23:629–652. https://doi.org/10.1097/01.WCB.0000073905.87928.6D
Wang X, Martindale JL, Liu Y, Holbrook NJ (1998) The cellular response to oxidative stress: influences of mitogen-activated protein kinase signalling pathways on cell survival. Biochem J 333(Pt 2):291–300. https://doi.org/10.1042/bj3330291
Wu H, Wu T, Xu X, Wang J, Wang J (2011) Iron toxicity in mice with collagenase-induced intracerebral hemorrhage. J Cereb Blood Flow Metab: Off J Int Soc Cereb Blood Flow Metab 31:1243–1250. https://doi.org/10.1038/jcbfm.2010.209
Yang G et al (2016) A combination of serum iron, ferritin and transferrin predicts outcome in patients with intracerebral hemorrhage. Sci Rep 6:21970. https://doi.org/10.1038/srep21970
Zhang L et al (2014) Autophagy suppression by exercise pretreatment and p38 inhibition is neuroprotective in cerebral ischemia. Brain Res 1587:127–132. https://doi.org/10.1016/j.brainres.2014.08.067
Zhao Y, Rempe DA (2011) Prophylactic neuroprotection against stroke: low-dose, prolonged treatment with deferoxamine or deferasirox establishes prolonged neuroprotection independent of HIF-1 function. J Cereb Blood Flow Metab: Off J Int Soc Cereb Blood Flow Metab 31:1412–1423. https://doi.org/10.1038/jcbfm.2010.230
Ziai WC (2013) Hematology and inflammatory signaling of intracerebral hemorrhage. Stroke 44:S74–S78. https://doi.org/10.1161/STROKEAHA.111.000662
Zille M, Karuppagounder SS, Chen Y, Gough PJ, Bertin J, Finger J, Milner TA, Jonas EA, Ratan RR (2017) Neuronal Death After Hemorrhagic Stroke In Vitro and In Vivo Shares Features of Ferroptosis and Necroptosis. Stroke 48:1033–1043. https://doi.org/10.1161/STROKEAHA.116.015609
Acknowledgments
The authors would like thanks to Professor Hideko Nagasawa and Dr. Tasuku Hirayama (Pharmaceutical and Medicinal Chemistry, Gifu Pharmaceutical University) for providing Si-RhoNox-1, a specific fluorescence probe to detect Fe2+.
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TI carried out most of the experiments and wrote the paper. HM and TS assisted in the establishment of the ICH mice model. ST performed the behavioral experiments. HM supported the cell counting assessment in immunostaining analysis. TI, SN, MS, and HH designed all the experiments and performed the data analysis. SN, MS, and HH gave advice and supervised the experimental works and helped to draft the manuscript. All authors discussed the results and implications and commented on the manuscript at all stages. All data were generated in-house and that no paper mill was used.
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All animal experiments performed in this study were approved by the animal experiment committees of Gifu Pharmaceutical University.
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Imai, T., Tsuji, S., Matsubara, H. et al. Deferasirox, a trivalent iron chelator, ameliorates neuronal damage in hemorrhagic stroke models. Naunyn-Schmiedeberg's Arch Pharmacol 394, 73–84 (2021). https://doi.org/10.1007/s00210-020-01963-6
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DOI: https://doi.org/10.1007/s00210-020-01963-6