Skip to main content

Advertisement

SpringerLink
Log in

Role of hepcidin and its downstream proteins in early brain injury after experimental subarachnoid hemorrhage in rats

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Early brain injury (EBI) is a major cause of mortality from subarachnoid hemorrhage (SAH). We aimed to study the pathophysiology of EBI and explore the role of hepcidin, a protein involved in iron homeostatic regulation, and its downstream proteins. One hundred and thirty-two male Sprague–Dawley rats were assigned into groups (n = 24/group): sham, SAH, SAH + hepcidin, SAH + hepcidin-targeting small interfering ribonucleic acid (siRNA), and SAH + scramble siRNA. Three hepcidin-targeting siRNAs and one scramble siRNA for hepcidin were injected 24 h before hemorrhage induction, and hepcidin protein was injected 30 min before induction. The rats were neurologically evaluated at 24 h and euthanized at 72 h. Hepcidin, ferroportin-1, and ceruloplasmin protein expression were measured by immunohistochemistry and Western blotting. Brain water content, blood–brain barrier (BBB) leakage, non-heme tissue iron and Garcia scale were evaluated. Hepcidin expression increased in the cerebral cortex and hippocampus after experimental SAH (P < 0.05 compared to sham), while ferroportin-1 and ceruloplasmin decreased (P < 0.05). Hepcidin injection lowered the expression of ferroportin-1 and ceruloplasmin further but siRNA reduced the levels of hepcidin (P < 0.05 compared to SAH) resulting in recovery of ferroportin-1 and ceruloplasmin levels. Apoptosis was increased in SAH rats compared to sham (P < 0.05) and increased slightly more by hepcidin, but decreased by siRNA (P < 0.05 compared to SAH). SAH rats had lower neurological scores, high brain water content, BBB permeability, and non-heme tissue iron (P < 0.05). In conclusion, downregulation of ferroportin-1 and ceruloplasmin caused by hepcidin enhanced iron-dependent oxidative damage and may be the potential mechanism of SAH.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Cahill J, Zhang JH (2009) Subarachnoid hemorrhage: is it time for a new direction? Stroke 40(3 Suppl):S86–S87. doi:10.1161/strokeaha.108.533315

    Article  PubMed  Google Scholar 

  2. Ostrowski RP, Colohan AR, Zhang JH (2006) Molecular mechanisms of early brain injury after subarachnoid hemorrhage. Neurol Res 28(4):399–414. doi:10.1179/016164106x115008

    Article  CAS  PubMed  Google Scholar 

  3. Matz PG, Fujimura M, Chan PH (2000) Subarachnoid hemolysate produces DNA fragmentation in a pattern similar to apoptosis in mouse brain. Brain Res 858(2):312–319

    Article  CAS  PubMed  Google Scholar 

  4. He Z, Ostrowski RP, Sun X, Ma Q, Huang B, Zhan Y, Zhang JH (2012) CHOP silencing reduces acute brain injury in the rat model of subarachnoid hemorrhage. Stroke 43(2):484–490. doi:10.1161/strokeaha.111.626432

    Article  PubMed  Google Scholar 

  5. Park CH, Valore EV, Waring AJ, Ganz T (2001) Hepcidin, a urinary antimicrobial peptide synthesized in the liver. J Biol Chem 276(11):7806–7810. doi:10.1074/jbc.M008922200

    Article  CAS  PubMed  Google Scholar 

  6. Oliveira SJ, de Sousa M, Pinto JP (2011) ER stress and iron homeostasis: a new frontier for the UPR. Biochem Res Int 2011:896474. doi:10.1155/2011/896474

    Article  PubMed  Google Scholar 

  7. Nemeth E, Tuttle MS, Powelson J, Vaughn MB, Donovan A, Ward DM, Ganz T, Kaplan J (2004) Hepcidin regulates cellular iron efflux by binding to ferroportin and inducing its internalization. Science 306(5704):2090–2093. doi:10.1126/science.1104742

    Article  CAS  PubMed  Google Scholar 

  8. Li L, Holscher C, Chen BB, Zhang ZF, Liu YZ (2011) Hepcidin treatment modulates the expression of divalent metal transporter-1, ceruloplasmin, and ferroportin-1 in the rat cerebral cortex and hippocampus. Biol Trace Elem Res 143(3):1581–1593. doi:10.1007/s12011-011-8967-3

    Article  CAS  PubMed  Google Scholar 

  9. Pigeon C, Ilyin G, Courselaud B, Leroyer P, Turlin B, Brissot P, Loreal O (2001) A new mouse liver-specific gene, encoding a protein homologous to human antimicrobial peptide hepcidin, is overexpressed during iron overload. J Biol Chem 276(11):7811–7819. doi:10.1074/jbc.M008923200

    Article  CAS  PubMed  Google Scholar 

  10. Frazer DM, Anderson GJ (2003) The orchestration of body iron intake: how and where do enterocytes receive their cues? Blood Cells Mol Dis 30(3):288–297

    Article  CAS  PubMed  Google Scholar 

  11. Qian ZM, Wang Q (1998) Expression of iron transport proteins and excessive iron accumulation in the brain in neurodegenerative disorders. Brain Res Brain Res Rev 27(3):257–267

    Article  CAS  PubMed  Google Scholar 

  12. Klomp LW, Farhangrazi ZS, Dugan LL, Gitlin JD (1996) Ceruloplasmin gene expression in the murine central nervous system. J Clin Invest 98(1):207–215. doi:10.1172/jci118768

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Wang Z, Meng CJ, Shen XM et al (2012) Potential contribution of hypoxia-inducible factor-1alpha, aquaporin-4, and matrix metalloproteinase-9 to blood-brain barrier disruption and brain edema after experimental subarachnoid hemorrhage. J Mol Neurosci 48(1):273–280. doi:10.1007/s12031-012-9769-6

    Article  CAS  PubMed  Google Scholar 

  14. Chen Z, Gao C, Hua Y, Keep RF, Muraszko K, Xi G (2011) Role of iron in brain injury after intraventricular hemorrhage. Stroke 42(2):465–470. doi:10.1161/strokeaha.110.602755

    Article  PubMed  Google Scholar 

  15. Hockel K, Trabold R, Scholler K, Torok E, Plesnila N (2012) Impact of anesthesia on pathophysiology and mortality following subarachnoid hemorrhage in rats. Exp Transl Stroke Med 4(1):5. doi:10.1186/2040-7378-4-5

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Tsubokawa T, Solaroglu I, Yatsushige H, Cahill J, Yata K, Zhang JH (2006) Cathepsin and calpain inhibitor E64d attenuates matrix metalloproteinase-9 activity after focal cerebral ischemia in rats. Stroke 37(7):1888–1894. doi:10.1161/01.str.0000227259.15506.24

    Article  CAS  PubMed  Google Scholar 

  17. Desland FA, Afzal A, Warraich Z, Mocco J (2014) Manual versus automated rodent behavioral assessment: comparing efficacy and ease of bederson and garcia neurological deficit scores to an open field video-tracking system. J Cent Nerv Syst Dis 6:7–14. doi:10.4137/JCNSD.S13194

    PubMed  PubMed Central  Google Scholar 

  18. Wang SM, Fu LJ, Duan XL, Crooks DR, Yu P, Qian ZM, Di XJ, Li J, Rouault TA, Chang YZ (2010) Role of hepcidin in murine brain iron metabolism. Cell Mol Life Sci 67(1):123–133. doi:10.1007/s00018-009-0167-3

    Article  CAS  PubMed  Google Scholar 

  19. Horky LL, Pluta RM, Boock RJ, Oldfield EH (1998) Role of ferrous iron chelator 2,2′-dipyridyl in preventing delayed vasospasm in a primate model of subarachnoid hemorrhage. J Neurosurg 88(2):298–303. doi:10.3171/jns.1998.88.2.0298

    Article  CAS  PubMed  Google Scholar 

  20. Chi SI, Wang CK, Chen JJ, Chau LY, Lin TN (2000) Differential regulation of H- and L-ferritin messenger RNA subunits, ferritin protein and iron following focal cerebral ischemia-reperfusion. Neuroscience 100(3):475–484

    Article  CAS  PubMed  Google Scholar 

  21. Gitlin JD (1998) Aceruloplasminemia. Pediatr Res 44(3):271–276. doi:10.1203/00006450-199809000-00001

    Article  CAS  PubMed  Google Scholar 

  22. Jomova K, Vondrakova D, Lawson M, Valko M (2010) Metals, oxidative stress and neurodegenerative disorders. Mol Cell Biochem 345(1–2):91–104. doi:10.1007/s11010-010-0563-x

    Article  CAS  PubMed  Google Scholar 

  23. Ong WY, Halliwell B (2004) Iron, atherosclerosis, and neurodegeneration: a key role for cholesterol in promoting iron-dependent oxidative damage? Ann N Y Acad Sci 1012:51–64

    Article  CAS  PubMed  Google Scholar 

  24. Ghribi O, Golovko MY, Larsen B, Schrag M, Murphy EJ (2006) Deposition of iron and beta-amyloid plaques is associated with cortical cellular damage in rabbits fed with long-term cholesterol-enriched diets. J Neurochem 99(2):438–449. doi:10.1111/j.1471-4159.2006.04079.x

    Article  CAS  PubMed  Google Scholar 

  25. Chan PH (2001) Reactive oxygen radicals in signaling and damage in the ischemic brain. J Cereb Blood Flow Metab 21(1):2–14. doi:10.1097/00004647-200101000-00002

    Article  CAS  PubMed  Google Scholar 

  26. Yeo JE, Kang SK (2007) Selenium effectively inhibits ROS-mediated apoptotic neural precursor cell death in vitro and in vivo in traumatic brain injury. Biochim Biophys Acta 1772(11–12):1199–1210. doi:10.1016/j.bbadis.2007.09.004

    Article  CAS  PubMed  Google Scholar 

  27. Nguyen TT, Cho SO, Ban JY, Kim JY, Ju HS, Koh SB, Song KS, Seong YH (2008) Neuroprotective effect of Sanguisorbae radix against oxidative stress-induced brain damage: in vitro and in vivo. Biol Pharm Bull 31(11):2028–2035

    Article  CAS  PubMed  Google Scholar 

  28. Matz PG, Fujimura M, Lewen A, Morita-Fujimura Y, Chan PH (2001) Increased cytochrome c-mediated DNA fragmentation and cell death in manganese-superoxide dismutase-deficient mice after exposure to subarachnoid hemolysate. Stroke 32(2):506–515

    Article  CAS  PubMed  Google Scholar 

  29. He Z, Ostrowski RP, Sun X, Ma Q, Tang J, Zhang JH (2012) Targeting C/EBP homologous protein with siRNA attenuates cerebral vasospasm after experimental subarachnoid hemorrhage. Exp Neurol 238(2):218–224. doi:10.1016/j.expneurol.2012.08.025

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This study was supported by the National Natural Science Foundation of China, National Clinical key specialty construction projects of China and funded project of Chongqing Science and Technology Commission.

Author information

Authors and Affiliations

  1. Department of Neurosurgery, First Affiliated Hospital of Chongqing Medical University, Chongqing, 400042, China

    Guanping Tan, Liu Liu, Zhaohui He, Jiujun Sun, Wenli Xing & Xiaochuan Sun

Authors
  1. Guanping Tan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Liu Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhaohui He
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jiujun Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wenli Xing
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xiaochuan Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Zhaohui He.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. The animal experimental ethics committee of Chongqing Medical University approved the experimental protocol.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Tan, G., Liu, L., He, Z. et al. Role of hepcidin and its downstream proteins in early brain injury after experimental subarachnoid hemorrhage in rats. Mol Cell Biochem 418, 31–38 (2016). https://doi.org/10.1007/s11010-016-2730-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-016-2730-1

Keywords

Search

Navigation