Abstract
Cyclooxygenase-2 (COX-2) has an established role in the pathogenesis of hypoxic-ischemic encephalopathy (HIE). In this study we sought to determine whether COX-2 was induced by asphyxia in newborn pigs, and whether neuronal COX-2 levels were affected by H2 treatment. Piglets were subjected to either 8 min of asphyxia or a more severe 20 min of asphyxia followed by H2 treatment (inhaling room air containing 2.1% H2 for 4 h). COX-2 immunohistochemistry was performed on brain samples from surviving piglets 24 h after asphyxia. The percentages of COX-2-immunopositive neurons were determined in cortical and subcortical areas. Only in piglets with more severe HIE, we observed significant, region-specific increases in neuronal COX-2 expression within the parietal and occipital cortices and in the CA3 hippocampal subfield. H2 treatment essentially prevented the increases in COX-2-immunopositive neurons. In the parietal cortex, the attenuation of COX-2 induction was associated with reduced 8'-hydroxy-2′-deoxyguanozine immunoreactivity and retained microglial ramifcation index, which are markers of oxidative stress and neuroinfiammation, respectively. This study demonstrates for the first time that asphyxia elevates neuronal COX-2 expression in a piglet HIE model. Neuronal COX-2 induction may play region-specific roles in brain lesion progression during HIE development, and inhibition of this response may contribute to the antioxidant/anti-infiammatory neuroprotective effects of H2 treatment.
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References
Thornberg E, Thiringer K, Odeback A, Milsom I. Birth asphyxia: incidence, clinical course and outcome in a Swedish population. Acta Paediatr 1995; 84: 927–32.
Pierrat V, Haouari N, Liska A, Thomas D, Subtil D, Truffert P. Prevalence, causes, and outcome at 2 years of age of newborn encephalopathy: population based study. Arch Dis Child Fetal Neonatal Ed 2005; 90: F257–61.
Vannucci RC, Perlman JM. Interventions for perinatal hypoxic-ischemic encephalopathy. Pediatrics 1997; 100: 1004–14.
Smith WL, Dewitt DL. Prostaglandin endoperoxide H synthases-1 and -2. Adv Immunol 1996; 62: 167–215.
Smith WL, DeWitt DL, Garavito RM. Cyclooxygenases: structural, cellular, and molecular biology. Annu Rev Biochem 2000; 69: 145–82.
Kaufmann WE, Andreasson KI, Isakson PC, Worley PF. Cyclooxygenases and the central nervous system. Prostaglandins 1997; 54: 601–24.
Yamagata K, Andreasson KI, Kaufmann WE, Barnes CA, Worley PF. Expression of a mitogen-inducible cyclooxygenase in brain neurons: regulation by synaptic activity and glucocorticoids. Neuron 1993; 11: 371–86.
Peri KG, Hardy P, Li DY, Varma DR, Chemtob S. Prostaglandin G/H synthase-2 is a major contributor of brain prostaglandins in the newborn. J Biol Chem 1995; 270: 24615–20.
Weerasinghe GR, Coon SL, Bhattacharjee AK, Harry GJ, Bosetti F. Regional protein levels of cytosolic phospholipase A2 and cyclooxygenase-2 in Rhesus monkey brain as a function of age. Brain Res Bull 2006; 69: 614–21.
Yang H, Chen C. Cyclooxygenase-2 in synaptic signaling. Curr Pharm Des 2008; 14: 1443–51.
Hahn T, Heinzel S, Plichta MM, Reif A, Lesch KP, Fallgatter AJ. Neurovascular coupling in the human visual cortex is modulated by cyclooxygenase-1 (COX-1) gene variant. Cereb Cortex 2011; 21: 1659–66.
Lecrux C, Toussay X, Kocharyan A, Fernandes P, Neupane S, Levesque M, et al. Pyramidal neurons are "neurogenic hubs" in the neurovascular coupling response to whisker stimulation. J Neurosci 2011; 31: 9836–47.
Strauss KI, Barbe MF, Marshall RM, Raghupathi R, Mehta S, Narayan RK. Prolonged cyclooxygenase-2 induction in neurons and glia following traumatic brain injury in the rat. J Neurotrauma 2000; 17: 695–711.
Dash PK, Mach SA, Moore AN. Regional expression and role of cyclooxygenase-2 following experimental traumatic brain injury. J Neurotrauma 2000; 17: 69–81.
Domoki F, Veltkamp R, Thrikawala N, Robins G, Bari F, Louis TM, et al. Ischemia-reperfusion rapidly increases COX-2 expression in piglet cerebral arteries. Am J Physiol 1999; 277: H1207–14.
Dégì R, Bari F, Thrikawala N, Beasley TC, Thore C, Louis TM, et al. Effects of anoxic stress on prostaglandin H synthase isoforms in piglet brain. Dev Brain Res 1998; 107: 265–76.
Seibert K, Zhang Y, Leahy K, Hauser S, Masferrer J, Perkins W, et al. Pharmacological and biochemical demonstration of the role of cyclooxygenase 2 in inflammation and pain. Proc Natl Acad Sci U S A 1994; 91: 12013–7.
Choi SH, Aid S, Bosetti F. The distinct roles of cyclooxygenase-1 and -2 in neuroinflammation: implications for translational research. Trends Pharmacol Sci 2009; 30: 174–81.
Fathali N, Ostrowski RP, Lekic T, Jadhav V, Tong W, Tang J, et al. Cyclooxygenase-2 inhibition provides lasting protection against neonatal hypoxic-ischemic brain injury. Crit Care Med 2010; 38: 572–8.
Toti P DFC, Schürfeld K, Stumpo M, Bartolommei S, Lombardi A, Petraglia E, Buonocore G. Cyclooxygenase-2 immunoreactivity in the ischemic neonatal human brain. An autopsy study. J Submicrosc Cytol Pathol 2001; 33: 245–9.
Dobbing J, Sands J. Comparative aspects of the brain growth spurt. Early HumDev 1979; 3: 79–83.
Olah O, Nemeth I, Toth-Szuki V, Bari F, Domoki F. Regional differences in the neuronal expression of cyclooxygenase-2 (COX-2) in the newborn pig brain. Acta Histochem Cytochem 2012; 45: 187–92.
Nemeth J, Toth-Szuki V, Varga V, Kovacs V, Remzso G, Domoki F. Molecular hydrogen affords neuroprotection in a translational piglet model of hypoxic-ischemic encephalopathy. J Physiol Pharmacol 2016; 67: 677–89.
Domoki F, Olah O, Zimmermann A, Nemeth I, Toth-Szuki V, Hugyecz M, et al. Hydrogen is neuroprotective and preserves cerebrovascular reactivity in asphyxiated newborn pigs. Pediatr Res 2010; 68: 387–92.
Ohsawa I, Ishikawa M, Takahashi K, Watanabe M, Nishimaki K, Yamagata K, et al. Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat Med 2007; 13: 688–94.
Cai J, Kang Z, Liu K, Liu W, Li R, Zhang JH, et al. Neuroprotective effects of hydrogen saline in neonatal hypoxia–ischemia rat model. Brain Res 2009; 1256: 129–37.
Cai J, Kang Z, Liu WW, Luo X, Qiang S, Zhang JH, et al. Hydrogen therapy reduces apoptosis in neonatal hypoxia–ischemia rat model. Neurosci Lett 2008; 441: 167–72.
Fu Y, Ito M, Fujita Y, Ito M, Ichihara M, Masuda A, et al. Molecular hydrogen is protective against 6-hydroxydopamine-induced nigrostriatal degeneration in a rat model of Parkinson's disease. Neurosci Lett 2009; 453: 81–5.
Qu J, Gan YN, Xie KL, Liu WB, Wang YF, Hei RY, et al. Inhalation of hydrogen gas attenuates ouabain-induced auditory neuropathy in gerbils. Acta Pharmacol Sin 2012; 33: 445–51.
Nagata K, Nakashima-Kamimura N, Mikami T, Ohsawa I, Ohta S. Consumption of molecular hydrogen prevents the stress-induced impairments in hippocampus-dependent learning tasks during chronic physical restraint in mice. Neuropsychopharmacology 2008; 34: 501–8.
Iketani M, Ohsawa I. Molecular hydrogen as a neuroprotective agent. Curr Neuropharmacol 2017; 15: 324–31.
Olah O, Toth-Szuki V, Temesvari P, Bari F, Domoki F. Delayed neurovascular dysfunction is alleviated by hydrogen in asphyxiated newborn pigs. Neonatology 2013; 104: 79–86.
Faulkner S, Bainbridge A, Kato T, Chandrasekaran M, Kapetanakis AB, Hristova M, et al. Xenon augmented hypothermia reduces early lactate/N-acetylaspartate and cell death in perinatal asphyxia. Ann Neurol 2011; 70: 133–50.
Randall GCB. The relationship of arterial blood pH and pCO2 to the viability of the newborn piglet. Can J Comp Med 1971; 35: 141–6.
Niwa K, Araki E, Morham SG, Ross ME, Iadecola C. Cyclooxygenase-2 contributes to functional hyperemia in whisker-barrel cortex. J Neurosci 2000; 20: 763–70.
Kaltschmidt B, Linker RA, Deng J, Kaltschmidt C. Cyclooxygenase-2 is a neuronal target gene of NF-kappaB. BMC Mol Biol 2002; 3: 16.
Hou YN, Vlaskovska M, Cebers G, Kasakov L, Liljequist S, Terenius L. A μ-receptor opioid agonist induces AP-1 and NF-κB transcription factor activity in primary cultures of rat cortical neurons. Neurosci Lett 1996; 212: 159–62.
Cadet P, Rasmussen M, Zhu W, Tonnesen E, Mantione KJ, Stefano GB. Endogenous morphinergic signaling and tumor growth. Front Biosci 2004; 9: 3176–86.
Gupta SC, Sundaram C, Reuter S, Aggarwal BB. Inhibiting NF-κB activation by small molecules as a therapeutic strategy. Biochim Biophys Acta, Gene Regul Mech 2010; 1799: 775–87.
Roka A, Kelen D, Halasz J, Beko G, Azzopardi D, Szabo M. Serum S100B and neuron-specific enolase levels in normothermic and hypothermic infants after perinatal asphyxia. Acta Paediatr 2012; 101: 319–23.
Tooley J, Eagle R, Satas S, Thoresen M. Significant head cooling can be achieved while maintaining normothermia in the newborn piglet. Arch Dis Child Fetal Neonatal Ed 2005; 90: F262–F6.
Xin Y, Liu H, Zhang P, Chang L, Xie K. Molecular hydrogen inhalation attenuates postoperative cognitive impairment in rats. NeuroReport 2017; 28: 694–700.
Hayashida K, Sano M, Kamimura N, Yokota T, Suzuki M, Maekawa Y, et al. H2 gas improves functional outcome after cardiac arrest to an extent comparable to therapeutic hypothermia in a rat model. J Am Heart Assoc 2012; 1: e003459. doi: 10.1161/JAHA.112.003459.
Park TS, Gidday JM, Gonzales E. Local cerebral blood flow response to locally infused 2-chloroadenosine during hypotension in piglets. Dev Brain Res 1991; 61: 73–7.
Gidday JM, Park TS. Effect of 2-chloroadenosine on cerebrovascular reactivity to hypercapnia in newborn pig. J Cereb Blood Flow Metab 1992; 12: 656–63.
Ruth VJ, Park TS, Gonzales ER, Gidday JM. Adenosine and cerebrovascular hyperemia during insulin-induced hypoglycemia in newborn piglet. Am J Physiol, Heart Circ Physiol 1993; 265: H1762–H8.
Ono H, Nishijima Y, Adachi N, Sakamoto M, Kudo Y, Kaneko K, et al. A basic study on molecular hydrogen (H2) inhalation in acute cerebral ischemia patients for safety check with physiological parameters and measurement of blood H2 level. Med Gas Res 2012; 2: 21.
Anjana Vaman VS, Tinu SK, Geetha CS, Lissy KK, Mohanan PV. Effect of fibrin glue on antioxidant defense mechanism, oxidative DNA damage and chromosomal aberrations. Toxicol Mech Methods 2013; 23: 500–8.
Villaño D, Vilaplana C, Medina S, Cejuela-Anta R, Martínez-Sanz JM, Gil P, et al. Effect of elite physical exercise by triathletes on seven catabolites of DNA oxidation. Free Radic Res 2015; 49: 973–83.
Hintsala HR, Jokinen E, Haapasaari KM, Moza M, RistimÄKi ARI, Soini Y, et al. Nrf2/Keap1 pathway and expression of oxidative stress lesions 8-hydroxy-2′-deoxyguanosine and nitrotyrosine in melanoma. Anticancer Res 2016; 36: 1497–506.
Nunomura A, Moreira PI, Castellani RJ, Lee HG, Zhu X, Smith MA, et al. Oxidative damage to RNA in aging and neurodegenerative disorders. Neurotox Res 2012; 22: 231–48.
Huang Q, Chen Z, Yang Y, Wang B, Li F, Zhan M. Content change and clinical significance of oxidation damage products 8-OHdG, 4-HNE and NTY in hemangioma tissue. Int J Clin Exp Pathol 2016; 9: 8847–57.
Campo GM, Avenoso A, Campo S, D'Ascola A, Traina P, Samà D, et al. The antioxidant effect exerted by TGF-1β-stimulated hyaluronan production reduced NF-kB activation and apoptosis in human fibroblasts exposed to FeSo4 plus ascorbate. Mol Cell Biochem 2008; 311: 167–77.
Hoffmann A, Levchenko A, Scott ML, Baltimore D. The IκB-NF-κB signaling module: temporal control and selective gene activation. Science 2002; 298: 1241–5.
O'Neill LAJ, Kaltschmidt C. NF-kB: a crucial transcription factor for glial and neuronal cell function. Trends Neurosci 1997; 20: 252–8.
Salminen A, Liu PK, Hsu CY. Alteration of transcription factor binding activities in the ischemic rat-brain. Biochem Biophys Res Commun 1995; 212: 939–44.
Shi Y, Wang G, Li J, Yu W. Hydrogen gas attenuates sevoflurane neurotoxicity through inhibiting nuclear factor kappa-light-chain-enhancer of activated B cells signaling and proinflammatory cytokine release in neonatal rats. Neuroreport 2017; 28: 1170–5.
Kaltschmidt C, Kaltschmidt B, Lannes-Vieira J, Kreutzberg GW, Wekerle H, Baeuerle PA, et al. Transcription factor NF-κB is activated in microglia during experimental autoimmune encephalomyelitis. J Neuroimmunol 1994; 55: 99–106.
Indo HP, Davidson M, Yen HC, Suenaga S, Tomita K, Nishii T, et al. Evidence of ROS generation by mitochondria in cells with impaired electron transport chain and mitochondrial DNA damage. Mitochondrion 2007; 7: 106–18.
Phaniendra A, Jestadi DB, Periyasamy L. Free radicals: properties, sources, targets, and their implication in various diseases. Indian J Clin Biochem 2015; 30: 11–26.
Block ML, Zecca L, Hong JS. Microglia-mediated neurotoxicity: uncovering the molecular mechanisms. Nat Rev Neurosci 2007; 8: 57–69.
Domoki F, Perciaccante JV, Puskar M, Bari F, Busija DW. Cyclooxygenase-2 inhibitor NS398 preserves neuronal function after hypoxia/ischemia in piglets. Neuroreport 2001; 12: 4065–8.
Acknowledgements
This study was supported by grants from the Hungarian Brain Research Program (No KTIA_13_NAP-A-I/13) and from the EU-funded Hungarian grant EFOP-3.6.1-16-2016–00014. János NÉMETH was supported by the “Nemzeti Tehetség Program” of the “Emberi Erőforrás Támogatáskezelő” from the Hungarian Ministry of Human Capacities.
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Varga, V., Németh, J., Oláh, O. et al. Molecular hydrogen alleviates asphyxia-induced neuronal cyclooxygenase-2 expression in newborn pigs. Acta Pharmacol Sin 39, 1273–1283 (2018). https://doi.org/10.1038/aps.2017.148
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DOI: https://doi.org/10.1038/aps.2017.148
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