Tumor Necrosis Factor-α and Nuclear Factor Kappa-β Expression in Rats Following Transient Focal Cerebral Ischemia Reperfusion

Hiba A Awooda^1, , Gihan M Sharara², Nepton Soltani³ and Amal M Saeed⁴

¹Department of Physiology, Faculty of Medicine, Alneelain University, Khartoum, Sudan

²Department of Biochemistry, Faculty of Medicine and Heath Sciences, Alexandria University, Alexandria, Egypt

³Department of Physiology, Faculty of Medicine, Hormozgan University of Medical Science, Bandar Abbas, Iran

⁴Department of Physiology – Faculty of Medicine–University of Khartoum – Khartoum, Sudan

Cite this paper:
Hiba A Awooda, Gihan M Sharara, Nepton Soltani and Amal M Saeed. Tumor Necrosis Factor-α and Nuclear Factor Kappa-β Expression in Rats Following Transient Focal Cerebral Ischemia Reperfusion. International Journal of Clinical and Experimental Neurology. 2014; 2(2):40-46. doi: 10.12691/ijcen-2-2-4

Abstract

Ischemic stroke usually initiates inflammation that potentiates neuronal death. The aim of this study was to evaluate the role of TNF-α and NF- қB in rats subjected to transient cerebral ischemia and to correlate their levels with the resulting of neurological deficits. Experimental procedures were performed on 30 adult male Wistar rats. In fifteen rats transient focal cerebral ischemia was induced by occlusion of the left common carotid artery (CCA) for 30 minutes followed by reperfusion for 24 hours (test group). Another 15 rats underwent the surgery at the same neck region without occlusion of CCA and served as a control group. Neurobehavioral assessments were evaluated. TNF-α was measured in the serum and brain tissue using ELISA method, and the expression of NF-қβ was done via western blotting as well. TNF-α concentration in both serum and brain tissue in the test group were significantly higher than control group (P < 0.001). The expression of NF- қB in the test group was significantly higher than control group (P < 0.001). Neurological deficit of the test group correlated negatively with both NF-қβ and TNF-α. Another positive correlation found between NF-қβ of the test group with the brain tissue and serum TNF-α. From the results of this study we can concluded that TNF-α and NF-қβ were significantly expressed in the affected brain tissue following cerebral ischemia/reperfusion in rats, with demonstration of a direct relationship between this inflammatory biomarkers and the consequent neurological deficits.

Keywords:
cerebral ischemia/reperfusion NF-қβ TNF-α

This work is licensed under a Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

References:

[1]	Bi Q, Wang T, Zhang W: Frequency and etiological diagnosis of ischemic stroke in Chinese young adults. Neurol Res 2012, 34: 354-358.
[2]	Donnan GA: The 2007 Feinberg lecture: a new road map for neuroprotection. Stroke 2008, 39: 242.
[3]	Deb P, Sharma S, Hassan KM: Pathophysiologic mechanisms of acute ischemic stroke: An overview with emphasis on therapeutic significance beyond thrombolysis. Pathophysiology 2010, 17: 197-218.
[4]	Durukan A, Tatlisumak T: Acute ischemic stroke: overview of major experimental rodent models, pathophysiology, and therapy of focal cerebral ischemia. Pharmacol Biochem Behav 2007, 87: 179-197.
[5]	Strickland I, Ghosh S: Use of cell permeable NBD peptides for suppression of inflammation. Ann Rheum Dis 2006, 65 Suppl 3: iii75-82.
[6]	Emsley HC, Tyrrell PJ: Inflammation and infection in clinical stroke. J Cereb Blood Flow Metab 2002, 22: 1399-1419.
[7]	Watters O, O'Connor JJ: A role for tumor necrosis factor-alpha in ischemia and ischemic preconditioning. J Neuroinflammation 2011, 8: 87.
[8]	Desai A, Singh N, Raghubir R: Neuroprotective potential of the NF-kappaB inhibitor peptide IKK-NBD in cerebral ischemia-reperfusion injury. Neurochem Int 2010, 57: 876-883.
[9]	Tak PP, Firestein GS: NF-kappaB: a key role in inflammatory diseases. J Clin Invest 2001, 107: 7-11.
[10]	Gidday JM, Gasche YG, Copin JC, Shah AR, Perez RS, Shapiro SD, Chan PH, Park TS: Leukocyte-derived matrix metalloproteinase-9 mediates blood-brain barrier breakdown and is proinflammatory after transient focal cerebral ischemia. Am J Physiol Heart Circ Physiol 2005, 289: H558-568.
[11]	Hayden MS, Ghosh S: Signaling to NF-kappaB. Genes Dev 2004, 18: 2195-2224.
[12]	Albensi BC, Mattson MP: Evidence for the involvement of TNF and NF-kappaB in hippocampal synaptic plasticity. Synapse 2000, 35: 151-159.
[13]	Ridder DA, Schwaninger M: NF-kappaB signaling in cerebral ischemia. Neuroscience 2009, 158: 995-1006.
[14]	Liu Y, Zhang XJ, Yang CH, Fan HG: Oxymatrine protects rat brains against permanent focal ischemia and downregulates NF-kappaB expression. Brain Res 2009, 1268: 174-180.
[15]	Keefer LK, Garland WA, Oldfield NF, Swagzdis JE, Mico BA: Inhibition of N-nitrosodimethylamine metabolism in rats by ether anesthesia. Cancer Res 1985, 45: 5457-5460.
[16]	Renolleau S, Aggoun-Zouaoui D, Ben-Ari Y, Charriaut-Marlangue C: A model of transient unilateral focal ischemia with reperfusion in the P7 neonatal rat: morphological changes indicative of apoptosis. Stroke 1998, 29: 1454-1460; discussion 1461.
[17]	Kuluz JW, Prado RJ, Dietrich WD, Schleien CL, Watson BD: The effect of nitric oxide synthase inhibition on infarct volume after reversible focal cerebral ischemia in conscious rats. Stroke 1993, 24: 2023-2029.
[18]	Furuya K, Zhu L, Kawahara N, Abe O, Kirino T: Differences in infarct evolution between lipopolysaccharide-induced tolerant and nontolerant conditions to focal cerebral ischemia. J Neurosurg 2005, 103: 715-723.
[19]	Star RA: Treatment of acute renal failure. Kidney Int 1998, 54: 1817-1831.
[20]	Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 1951, 193: 265-275.
[21]	Leist M, Gantner F, Naumann H, Bluethmann H, Vogt K, Brigelius-Flohe R, Nicotera P, Volk HD, Wendel A: Tumor necrosis factor-induced apoptosis during the poisoning of mice with hepatotoxins. Gastroenterology 1997, 112: 923-934.
[22]	Bers G, Garfin D: Protein and nucleic acid blotting and immunobiochemical detection. BioTechniques 1985, 3: 276-288.
[23]	Hosomi N, Ban CR, Naya T, Takahashi T, Guo P, Song XY, Kohno M: Tumor necrosis factor-alpha neutralization reduced cerebral edema through inhibition of matrix metalloproteinase production after transient focal cerebral ischemia. J Cereb Blood Flow Metab 2005, 25: 959-967.
[24]	Murakami Y, Saito K, Hara A, Zhu Y, Sudo K, Niwa M, Fujii H, Wada H, Ishiguro H, Mori H, Seishima M: Increases in tumor necrosis factor-alpha following transient global cerebral ischemia do not contribute to neuron death in mouse hippocampus. J Neurochem 2005, 93: 1616-1622.
[25]	Sriram K, O'Callaghan JP: Divergent roles for tumor necrosis factor-alpha in the brain. J Neuroimmune Pharmacol 2007, 2: 140-153.
[26]	Clausen BH, Lambertsen KL, Babcock AA, Holm TH, Dagnaes-Hansen F, Finsen B: Interleukin-1beta and tumor necrosis factor-alpha are expressed by different subsets of microglia and macrophages after ischemic stroke in mice. J Neuroinflammation 2008, 5: 46.
[27]	Ekdahl CT, Kokaia Z, Lindvall O: Brain inflammation and adult neurogenesis: the dual role of microglia. Neuroscience 2009, 158: 1021-1029.
[28]	Haddad M, Rhinn H, Bloquel C, Coqueran B, Szabo C, Plotkine M, Scherman D, Margaill I: Anti-inflammatory effects of PJ34, a poly(ADP-ribose) polymerase inhibitor, in transient focal cerebral ischemia in mice. Br J Pharmacol 2006, 149: 23-30.
[29]	Lotocki G, Alonso OF, Dietrich WD, Keane RW: Tumor necrosis factor receptor 1 and its signaling intermediates are recruited to lipid rafts in the traumatized brain. J Neurosci 2004, 24: 11010-11016.
[30]	Marchetti L, Klein M, Schlett K, Pfizenmaier K, Eisel UL: Tumor necrosis factor (TNF)-mediated neuroprotection against glutamate-induced excitotoxicity is enhanced by N-methyl-D-aspartate receptor activation. Essential role of a TNF receptor 2-mediated phosphatidylinositol 3-kinase-dependent NF-kappa B pathway. J Biol Chem 2004, 279: 32869-32881.
[31]	Intiso D, Zarrelli MM, Lagioia G, Di Rienzo F, Checchia De Ambrosio C, Simone P, Tonali P, Cioffi Dagger RP: Tumor necrosis factor alpha serum levels and inflammatory response in acute ischemic stroke patients. Neurol Sci 2004, 24: 390-396.
[32]	Sairanen T, Carpen O, Karjalainen-Lindsberg ML, Paetau A, Turpeinen U, Kaste M, Lindsberg PJ: Evolution of cerebral tumor necrosis factor-alpha production during human ischemic stroke. Stroke 2001, 32: 1750-1758.
[33]	Tarkowski E, Rosengren L, Blomstrand C, Wikkelso C, Jensen C, Ekholm S, Tarkowski A: Intrathecal release of pro- and anti-inflammatory cytokines during stroke. Clin Exp Immunol 1997, 110: 492-499.
[34]	Mao DJ, Guo RY, Tang YC, Zang YH: [Expression of sCD40L in peripheral blood and NF-kappaBp65 in PBMC of patients with acute progressive cerebral infarction]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi 2011, 27: 177-179.
[35]	Jeong HJ, Choi IY, Kim MH, Kim HM, Moon PD, Hong JW, Kim SH: Chungsim-Yeunja-Tang decreases the inflammatory response in peripheral blood mononuclear cells from patients with cerebral infarction through an NF-kappaB dependent mechanism. J Neuroinflammation 2010, 7: 85.
[36]	Lou HY, Wei XB, Zhang B, Sun X, Zhang XM: Hydroxyethylpuerarin attenuates focal cerebral ischemia-reperfusion injury in rats by decreasing TNF-alpha expression and NF-kappaB activity. Yao Xue Xue Bao 2007, 42: 710-715.
[37]	Zheng JM, Chen XC, Lin M, Zhang J, Lin ZY, Zheng GY, Li KZ: [Mechanism of the reduction of cerebral ischemic-reperfusion injury through inhibiting the activity of NF-kappaB by propyl gallate]. Yao Xue Xue Bao 2011, 46: 158-164.