DOI: http://dx.doi.org/10.18203/2320-6012.ijrms20150323

Comparison of cerebrospinal fluid Cytochrome-c and Caspase-9 as biomarkers for newborns with hypoxic ischemic encephalopathy with non-asphyxiated babies and followup of these biomarkers after day 7

Supriya Kushwah, Ashok Kumar, Sriparna Basu, Sairam Krishnamurthy, Ashutosh Kumar

Abstract


Background: There are very less previous study for cytochrome–c and caspase-9, the key players in apoptotic cell death, in human newborns. The objective was to measure the level of cerebrospinal fluid biomarkers cytochrome –c and caspase -9 in newborns with hypoxic ischemic encephalopathy and comparison with clinically suspected sepsis controls and to compare these after 7 days.

Methods: We compared 50 hypoxic babies with 20 newborns with clinically suspected sepsis at median age of day-3 and 9 in cases and day-1 in controls.

Results: In the present study in sample-1 we observed a significant increase in the levels of cases cytochrome c (1.46 ± 0.71 ng⁄mL) and caspase- 9 (0.29 ± 0.27 ng⁄mL) when compared to controls cytochrome-c (1.02+0.27 ng⁄mL) and caspase -9 (0.13+0.16 ng⁄mL) with significant p-value of 0.001 and 0.009 respectively. In sample -1 Cytochrome-c, P- value was significant when compared stage –III (1.74 ± 0.68) with stage-I (0.82 ± 0.43) and stage –II (0.99 ± 0.18). Similarly in Caspas-9 P-value was significant when compared between stage-III (0.38 ± 0.30) with stage-I (0.11 ± 0.07). In sample -2 P- value was significant when compared stage –III (1.68 ± 0.50) with stage-I (1.01 ± 0.14) and stage –II (0.94 ± 0.38). Similarly in Caspas-9 P-value was significant when compared between stage-III (4.84 ± 2.44) with stage-I (0.13 ± 0.10) and stage –II (0.13 ± 0.11).

Conclusions: First time done in human newborns with asphyxia, showing that CSF Cytochrome- c and Caspase 9 increases significantly. In sample-2, the caspase 9 levels showed a further increase, whereas cytochrome c levels decreased from the sample 1 value indicating that neuroprotection time should be increased.

 


Keywords


Cytochrome-C, Caspase-9, Hypoxic ischemic encephalopathy, Newborns

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References


Whitelaw A, Thoresen M. Clinical trials of treatments after perinatal asphyxia. Curr Opin Pediatr. 2002;14:664–8.

Shankaran S, Laptook AR. Hypothermia as a treatment for birth asphyxia. Clin Obstet Gynecol. 2007;50:624 –35.

Gonzalez FF, Ferriero DM. Therapeutics for neonatal brain injury. Pharmacol Ther. 2008;120:43–53.

Lawn JE, Cousens S, Zupan J. Lancet neonatal survival steering team. Four million neonatal deaths: When? Where? Why? Lancet. 2005;365:891–900.

Eisenberg-Lerner A, Bialik S, Simon HU, Kimchi A. Life and death partners: apoptosis, autophagy and the cross-talk between them. Cell Death Differ. 2009;16:966–75.

Nelson, K. B., J. M. Dambrosia, et al. "Neonatal cytokines and coagulation factors in children with cerebral palsy." Ann Neurol. 1998;44(4):665-75.

Lubec B, Chiappe-Gutierrez M, Hoeger H, Kitzmueller E, Lubec G. Glucose transporters, hexokinase, and phosphofructokinase in brain of rats with perinatal asphyxia. Pediatr Res. 2000;47:84–8.

Rundgren M, Karlsson T, Nielsen N, Cronberg T, Johnsson P, Friberg H. Neuron specific enolase and S-100B as predictors of outcome after cardiac arrest and induced hypothermia. Resuscitation. 2009;80:784–9.

Imam SS, Gad GI, Atef SH, Shawky MA. Cord blood brain derived neurotrophic factor: diagnostic and prognostic marker in full term newborns

with perinatal asphyxia. Pak J Biol Sci. 2009;12:1498–504.

Gazzolo D, Abella R, Frigiola A, Giamberti A, Tina G, Nigro F, et al. Neuromarkers and unconventional biological fluids. J Matern Fetal Neonatal Med. 2010;23:66–9.

Bembea MM, Savage W, Strouse JJ, Schwartz JM, Graham E, Thompson CB, et al. Glial fibrillary acidic protein as a brain injury biomarker in children undergoing extracorporeal membrane oxygenation. Pediatr Crit Care Med. 2010;11:723–30.

Gao Y, Liang W, Hu X, Zhang W, Stetler RA, Vosler P, et al. Neuroprotection against hypoxic-ischemic brain injury by inhibiting the apoptotic protease activating factor-1 pathway. Stroke. 2010;41(1):166-72.

Feuerstein GZ, Wang XK, Barone EC. Inflammatory mediators of brain injury: the role of cytokines and chemokines in stroke and CNS diseases. In Cerebrovascular Diseases: Pathophysiology, Diagnosis Management (Ed. M.G. Ginsberg and J. Bogousslavsky) London. Blackwell Scientific Publications 1996; 507-531.

Northington FJ, Zelaya ME, O’Riordan DP, Blomgren K, Flock DL, Hagberg H, et al. Failure to complete apoptosis following neonatal hypoxia-ischemia manifests as “continuum” phenotype of cell death and occurs with multiple manifestations of mitochondrial dysfunction in rodent forebrain. Neuroscience. 2007;149:822–33.

Gitto E, Reiter RJ, Karbownik M, Tan DX, Gitto P, Barberi S, et al. Causes of oxidative stress in the pre- and perinatal period. Biol Neonate. 2002;81:146–57.

Dringen R, Pawlowski PG, Hirrlinger J. Peroxide detoxification by brain cells. J Neurosci Res. 2005;79:157–65.

Palmer CL, Cotton L, Henley JM. The molecular pharmacology and cell biology of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptors. Pharmacol Rev. 2005;57(2):253-77.

Numagami Y, Zubrow AB, Mishra OP. Delivoria-Papadopoulos M. Lipid free radical generation and brain cell membrane alteration following nitric oxide synthase inhibition during cerebral hypoxia in the newborn piglet. J Neurochem. 1997;69:1542–7.

Chen Y, Engidawork E, Loidl F, Dell’Anna E, Goiny M, Lubec G, et al. Short- and long-term effects of perinatal asphyxia on monoamine, amino acid and glycolysis product levels measured in the basal ganglia of the rat. Brain Res Dev Brain Res. 1997;104:19–30.

Bonfoco E, Krainc D, Ankarcrona M, Nicotera P, Lipton SA. Apoptosis and necrosis: two distinct events induced, respectively, by mild and intense insults with N-methyl-D-aspartate or nitric oxide/ superoxide in cortical cell cultures. Proc Natl Acad Sci USA. 1995;92(16):7162-6.

Holopainen IE, Lauren HB, Romppanen A, Lopez-Picon FR. Changes in neurofilament protein-immunoreactivity after kainic acid treatment of organotypic hippocampal slice cultures. J Neurosci Res. 200;66(4):620-9.

Brenner C, Cadiou H, Vieira HL, Zamzami N, Marzo I, Xie Z, et al. Bcl-2 and Bax regulate the channel activity of the mitochondrial adenine nucleotide translocator. Oncogene. 2000;19(3):329-36.

Capani F, Loidl CF, Aguirre F, Piehl L, Facorro G, Hager A, et al. Changes in reactive oxygen species (ROS) production in rat brain during global perinatal asphyxia: an ESR study. Brain Res. 2001;914:204–7.

Gazzolo D, Marinoni E, Iorio RD, Bruschettini M, Kornacka M, Lituania M, et al. Measurement of Urinary S100B Protein Concentrations for the Early Identification of Brain Damage in Asphyxiated Full-term Infant. Arch Pediatr Adolesc Med. 2003;157:1163-8.

Thornberg E, Thiringer K, Hagberg H, Kjellmer I. Neuron specific enolase in asphyxiated newborns: association with encephalopathy and cerebral function monitor trace. Arch Dis Child. 1995;72:39-42.

Lubec B, Marx M, Herrera-Marschitz M, Labudova O, Hoeger H, Gille L, et al. Decrease of heart protein kinase C and cyclin-dependent kinase precedes death in perinatal asphyxia of the rat. FASEB J. 1997;11:482–92.

Blennow M, Hagberg H, Rosengren L. Glial fi brillarycidic protein in the cerebrospinal fluid: a possible indicator of prognosis in full-term asphyxiated newborn infants? Pediatr Res. 1995;37:260-4.

Ferriero DM. Neonatal brain injury. N Engl J Med. 2004;351:1985–95.

Krajewski S, Krajewska M, Ellerby LM, Welsh K, Xie Z, Deveraux QL, et al. Release of caspase-9 from mitochondria during neuronal apoptosis and cerebral ischemia. Proc Natl Acad Sci USA 96. 1999;5752-7.

Hagberg H, Mallard C, Rousset CI, Xiaoyang W. Apoptotic mechanisms in the immature brain: involvement of mitochondria. J Child Neurol. 2009;24:1141–6.

Yuan J, Yankner BA. Apoptosis in the nervous system. Nature. 2000;407:802–9.

Kim P, Leckman JF, Mayes LC, Feldman R, Wang X, Swain JE. The plasticity of human maternal brain: Longitudinal changes in brain anatomy during the early postpartum period. Behavioral Neuroscience. 2010;124:695–700.

Hagberg H, Wilson MA, Matsushita H. PARP-1 gene disruption in mice preferentially protects males from perinatal brain injury. J Neurochem. 2004;90:1068–75.

Darwish RS, Amiridze N, Aarabi B. Nitrotyrosine as an oxidative stress marker: evidence for involvement in neurologic outcome in human traumatic brain injury. J Trauma. 2007;63(2):439-42.

Matsumori Y, Northington FJ, Hong SM, Kayama T, Sheldon RA, Vexler ZS, et al. Reduction of Caspase-8 and -9 Cleavage Is Associated With Increased c-FLIP and Increased Binding of Apaf-1 and Hsp70 After Neonatal Hypoxic/Ischemic Injury in Mice Overexpressing Hsp70. Stroke. 2006;37:507-12.

Chauvier D, Renolleau S, Holifanjaniaina S, Ankri S, Bezault M, Schwendimann L, et al. Targeting neonatal ischemic brain injury with a pentapeptide-based irreversible caspase inhibitor. Cell Death Dis. 2011;2:e203.

Saikumar P, Dong Z, Pate Y, Hall K, Hopfer U, Weinberg JM, et al. Role of hypoxia-induced Bax translocation and cytochrome c release in reoxygenation injury. Oncogene. 1998;17:3401–15.

Feng Y, Shi W, Huang M, LeBlanc MH. Oxypurinol administration fails to prevent hypoxic-ischemic brain injury in neonatal rats. Brain Res Bull. 2003;59:453-7.

Deng H, Dodson MW, Huang H, Guo M. The Parkinson's disease genes pink1 and parkin promote mitochondrial fission and/or inhibit fusion in Drosophila. Proc Natl Acad Sci USA. 2008;105(38):14503-8.