Abstract
The members of high-temperature requirement (HtrA) family are evolutionarily conserved serine proteases that combine a trypsin-like protease domain with at least one PDZ interaction domain. HtrA2, a special one, is mainly located in mitochondria and required for maintaining homeostasis. Once released into cytoplasm, HtrA2 contributes to apoptosis via caspase-dependent and -independent pathways. Accumulating evidence has showed its pro-apoptotic effect in cancers and central nervous system (CNS) diseases. However, the distribution and function of HtrA2 in CNS diseases remains to be further explored. To investigate HtrA2’s roles in the pathophysiology of intracerebral hemorrhage (ICH), an ICH rat model was established and assessed by behavioral tests. Western blot and immunohistochemistry revealed a remarkable up-regulation of HtrA2 surrounding the hematoma after ICH; and immunofluorescence showed HtrA2 was strikingly increased in neurons, but not in astrocytes and oligodendrocytes. Terminal deoxynucleotidyl transferase-mediated biotinylated-dUTP nick-end labeling staining suggested the involvement of HtrA2 in neuronal apoptosis after ICH. Additionally, HtrA2 co-localized with active-caspase-3 around the hematoma and the expression of active-caspase-3 was parallel with that of HtrA2 in a time-dependent manner. Furthermore, hemin was used to stimulus a neuronal cell line PC12 to mimic ICH model in vitro. We analyzed the relationship of HtrA2 with X-linked inhibitor of apoptosis protein (XIAP) in PC12 cells by Western blot, immunofluorescence and co-immunoprecipitation. The connection of HtrA2 with XIAP was strengthened in apoptotic cells after hemin treatment. Thus, we speculated that HtrA2 might exert an important function in regulating caspase-dependent neuronal apoptosis through interacting with XIAP following ICH.
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References
Aronowski J, Zhao X (2011) Molecular pathophysiology of cerebral hemorrhage: secondary brain injury. Stroke 42(6):1781–1786
Bradl M, Lassmann H (2010) Oligodendrocytes: biology and pathology. Acta Neuropathol 119(1):37–53
Broughton BR, Reutens DC, Sobey CG (2009) Apoptotic mechanisms after cerebral ischemia. Stroke 40(5):e331–e339
Cilenti L et al (2004) Regulation of HAX-1 anti-apoptotic protein by Omi/HtrA2 protease during cell death. J Biol Chem 279(48):50295–50301
Cilenti L et al (2005) Omi/HtrA2 protease mediates cisplatin-induced cell death in renal cells. Am J Physiol Renal Physiol 288(2):F371–F379
Clausen T, Southan C, Ehrmann M (2002) The HtrA family of proteases: implications for protein composition and cell fate. Mol Cell 10(3):443–455
Desideri E, Martins LM (2012) Mitochondrial stress signalling: HtrA2 and Parkinson’s disease. Int J Cell Biol 2012:607929
Flanagan L et al (2010) XIAP impairs Smac release from the mitochondria during apoptosis. Cell Death Dis 1:e49
Gong C et al (2001) Intracerebral hemorrhage-induced neuronal death. Neurosurgery, 48(4): 875–882 (discussion 882–883)
Hua Y et al (2002) Behavioral tests after intracerebral hemorrhage in the rat. Stroke 33(10):2478–2484
Ikram MA, Wieberdink RG, Koudstaal PJ (2012) International epidemiology of intracerebral hemorrhage. Curr Atheroscler Rep 14(4):300–306
Jellinger K (1977) Pathology of intracerebral hemorrhage. Zentralbl Neurochir 38(1):29–42
Karabiyikoglu M, Hua Y, Keep RF, Ennis SR, Xi G (2004) Intracerebral hirudin injection attenuates ischemic damage and neurologic deficits without altering local cerebral blood flow. J Cereb Blood Flow Metab 24(2):159–166
Karbowski M (2010) Mitochondria on guard: role of mitochondrial fusion and fission in the regulation of apoptosis. Adv Exp Med Biol 687:131–142
Kuninaka S et al (2005) The tumor suppressor WARTS activates the Omi/HtrA2-dependent pathway of cell death. Oncogene 24(34):5287–5298
Kuwana T, Newmeyer DD (2003) Bcl-2-family proteins and the role of mitochondria in apoptosis. Curr Opin Cell Biol 15(6):691–699
Levy YS, Streifler JY, Panet H, Melamed E, Offen D (2002) Hemin-induced apoptosis in PC12 and neuroblastoma cells: implications for local neuronal death associated with intracerebral hemorrhage. Neurotox Res 4(7–8):609–616
Liu HR et al (2005) Role of Omi/HtrA2 in apoptotic cell death after myocardial ischemia and reperfusion. Circulation 111(1):90–96
Liu QB et al (2010) The induction of reactive oxygen species and loss of mitochondrial Omi/HtrA2 is associated with S-nitrosoglutathione-induced apoptosis in human endothelial cells. Toxicol Appl Pharmacol 244(3):374–384
Mayer B, Oberbauer R (2003) Mitochondrial regulation of apoptosis. News Physiol Sci 18:89–94
Pallen MJ, Wren BW (1997) The HtrA family of serine proteases. Mol Microbiol 26(2):209–221
Porter AG, Janicke RU (1999) Emerging roles of caspase-3 in apoptosis. Cell Death Differ 6(2):99–104
Scott FL et al (2005) XIAP inhibits caspase-3 and -7 using two binding sites: evolutionarily conserved mechanism of IAPs. EMBO J 24(3):645–655
Sukumari-Ramesh S, Alleyne CH, Jr and Dhandapani KM, 2012 Astrocyte-specific expression of survivin after intracerebral hemorrhage in mice: a possible role in reactive gliosis? J Neurotrauma
Suzuki Y et al (2001) A serine protease, HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death. Mol Cell 8(3):613–621
Vande Walle L, Lamkanfi M, Vandenabeele P (2008) The mitochondrial serine protease HtrA2/Omi: an overview. Cell Death Differ 15(3):453–460
Xi G, Keep RF, Hoff JT (2006) Mechanisms of brain injury after intracerebral haemorrhage. Lancet Neurol 5(1):53–63
Xue M, Del Bigio MR (2000) Intracerebral injection of autologous whole blood in rats: time course of inflammation and cell death. Neurosci Lett 283(3):230–232
Xue M, Del Bigio MR (2003) Comparison of brain cell death and inflammatory reaction in three models of intracerebral hemorrhage in adult rats. J Stroke Cerebrovasc Dis 12(3):152–159
Yang S et al (2008) Effects of thrombin on neurogenesis after intracerebral hemorrhage. Stroke 39(7):2079–2084
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Huiqing Sun and Lei Li contributed equally to this work.
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Sun, H., Li, L., Zhou, F. et al. The member of high temperature requirement family HtrA2 participates in neuronal apoptosis after intracerebral hemorrhage in adult rats. J Mol Hist 44, 369–379 (2013). https://doi.org/10.1007/s10735-013-9489-4
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DOI: https://doi.org/10.1007/s10735-013-9489-4