Basic fibroblast growth factor alleviates brain injury following global ischemia reperfusion in rabbits

Basic fibroblast growth factor alleviates brain injury following global ischemia reperfusion in rabbits 1 first-author †E-mail:zmhz@hotmail.com; yuefengma@hotmail.com †E-mail:zmhz@hotmail.com; yuefengma@hotmail.com zmhz@hotmail.com 1 corresp ‡Corresponding Author ‡Corresponding Author †E-mail:zmhz@hotmail.com; yuefengma@hotmail.com †E-mail:zmhz@hotmail.com; yuefengma@hotmail.com zmhz@hotmail.com 1 1 1 1 2 2 Department of Emergency Medicine, Second Affiliated Hospital, Zhejiang University, School of Medicine, Hangzhou 310009, China Department of Emergency Medicine, Second Affiliated Hospital, Zhejiang University, School of Medicine, Hangzhou 310009, China Institute of Biological Engineering, Jinan University, Guangzhou 510632, China Institute of Biological Engineering, Jinan University, Guangzhou 510632, China The aim of this study was to explore the protective effect of basic fibroblast growth factor (bFGF) on brain injury following global ischemia reperfusion and its mechanisms. Brain injury following global ischemia was induced by four vessels occlusion and systemic hypotension. Twenty-four rabbits were randomized into three groups: group A, only dissection of vessels; group B, intravenous infusion of normal saline after reperfusion for 6 h; group C, 30 μg/kg bFGF injected intravenously at the onset of reperfusion, then infused with 10 μg/(kg·h) for 6 h. Serum neuron specific enolase (NSE), S-100B, tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1), interleukin-8 (IL-8) were measured before ischemia, 30 min after ischemia, 0.5, 1, 3, 6 h after reperfusion. Brain water content was determined and cerebral histopathological damages were compared. NSE and S-100B were increased 1 h after reperfusion and reached their peaks 6 h after reperfusion, but were much higher in group B than those in group C 3, 6 h after reperfusion. In groups B and C, TNF-α was increased after ischemia and IL-1 and IL-8 were increased significantly 0.5 h after reperfusion, then reached their peaks 6 h, 3 h, 6 h after reperfusion respectively. TNF-α and IL-8 at the time points of 1 h and 3 h and IL-1 at 3 h and 6 h in group C were correspondingly lower than those in group B. These indices in group A were nearly unchanged. There were less severe cerebral histopathological damages in group C compared with group B, but no difference in brain water content. It could be concluded that bFGF alleviates brain injury following global ischemia and reperfusion by down-regulating expression of inflammatory factors and inhibiting their activities. Basic fibroblast growth factor Brain injury Global ischemia Neuron specific enolase S-100B protein Tumor necrosis factor-α Interleukin INTRODUCTION 1 With increasing public knowledge on cardiopulmonary resuscitation (CPR) and improvements in emergent early response, these have resulted in higher percentage of patients with out-of-hospital cardiac arrest (CA) surviving to admission (Bunch et al., 2004 2004 ). The hospital discharge rate is only 2%~14%, with most patients being still left with poor neurological outcome (Snyder-Ramos and Bottiger, 2003 2003 ), due to persistent brain damage. Cerebral resuscitation plays a key role during the CPR process and remains a challenge. Recent studies showed basic fibroblast growth factor (bFGF) had neuro-protective effect against focal ischemic, excitotoxic and traumatic brain injury (Dietrich et al., 1996 1996 ; Bethel et al., 1997 1997 ; Himmelseher et al., 1997 1997 ; Yoshimura et al., 2003 2003 ). However the effect on global cerebral ischemia-reperfusion injury, following CA and CPR, has not been investigated. By creating an animal model of global ischemia and reperfusion, we investigated effects of bFGF on serum neuron specific enolase (NSE), S-100B, tumor necrosis factor-alpha (TNF-α), interleukin-1 (IL-1), interleukin-8 (IL-8), in order to explore the potential neuroprotection of bFGF in cardiopulmonary resuscitation and the possible mechanisms. MATERIALS AND METHODS 1 Animals and model 2 Cerebral ischemia-reperfusion injury following cardiac arrest was simulated by four vessels occlusion (2 carotids and 2 vertebral arteries) and systemic hypotension (Pomfy and Franko, 1999 1999 ). Twenty-four healthy rabbits (supplied by Experimental Animal Center, Medical Institute of Zhejiang Province) with mean body weight of 2.6 kg (2.2~3.0 kg), regardless of gender, were randomized into three groups: group A ( n =8), subjected to dissection of four vessels without occlusion; group B ( n =8), who underwent dissection and occlusion of vessels, infusion of normal saline (NS); group C ( n =8), who underwent dissection and occlusion of vessels, infusion of bFGF. The rabbits were anesthetized by slow intravenous injection of Diazepam (4 mg/kg), Haloperidol (0.8 mg/kg) and Fentanyl (16 μg/kg). The femoral artery was cannulated and connected to a transducer. Direct arterial blood pressure together with electrocardiograph (ECG), rectal temperature and partial pressure of end tidal carbon dioxide ( P ET,CO 2 ) were monitored using a multiple-parameters monitor (MB-526, USA). Rectal temperature was kept at about 38.5 °C with a heating system. Tracheotomy was performed and the rabbits were intubated, ventilated with an DH-150 animal ventilator (Medical Instrument Factory of Zhejiang University, China). Ventilator settings were adjusted to maintain P ET,CO 2 at 40 mmHg. Both common carotids and vertebral arteries were dissected and clamped, while hypotension was induced by exsanguinations via femoral arteries, mean blood pressure was kept at 50%~60% of basal value. After 30 min, the four vessels were unclamped and blood was transfused rapidly. In group B, a bolus of 5 ml NS was given intravenously, followed by a continuous infusion of NS at a rate of 5 ml/(kg·h). In group C, it was replaced by a bolus of 30 μg/kg bFGF (Recombinant bovine basic fibroblast growth factor, 36 micrograms/ampoule, Changchun Changsheng Gene Pharmaceutical Co. Ltd., China), then 10 μg/(kg·h) infused continuously, dissolved in the same volume of NS. Sample preparation and measurements 2 Three milliliters of blood samples were taken before occlusion of vessels (basal time), at the end of ischemia (0 h), 0.5, 1, 3, 6 h after reperfusion respectively and the same volume of NS was infused after sampling. All blood specimens were processed to serum, centrifugated at 3000 rpm for 10 min at room temperature, then stored in aliquots at −80 °C until batch evaluation. Serum NSE, S-100B, TNF-α, IL-1 and IL-8 were measured by enzyme linked immunosorbant assay on Opsys MR microplate reader (Dynex Inc., USA), according to the operating instructions. Kits for rabbit NSE and S-100B were supplied by Lifekey BioMeditech Inc. (USA), and those for TNF-α, IL-1, IL-8 by Sigma Inc. (USA). The animals were sacrificed by exsanguination 6 h after reperfusion. The right hemispheres were sampled and kept at 100 °C in a drier for 12 h, then the brain water content was determined by the difference between dry and wet weight. The left hemispheres were stored in 10% formalin. Histopathological sections were performed routinely, stained with hematoxylin and eosin (HE). Statistical analysis 2 All data were expressed as mean±standard deviation. Differences between groups or different time points were compared using one-way ANOVA and Tukey test (SigmaStat 2.03 software package was used). Correlation between serum NSE and S-100B was analyzed with linear regression. A P value of < 0.05 was considered as statistically significant. RESULTS 1 General animal characteristics 2 There were no differences in body weight, sexual ratio, and basal values of rectal temperature, arterial blood pressure and heart rate between the three groups. Compared with group A, arterial blood pressure in groups B and C increased rapidly after occlusion of four vessels, and was maintained at half of the basal value during exsanguination, decreased sequentially after reperfusion, ameliorated gradually after transfusion, then decreased again 1 h after reperfusion. Blood pressure at different time points in group C seemed slightly lower than that in group B, but the differences were not statistically significant. Changes in serum NSE and S-100B 2 As shown in Fig. 1 1 , serum NSE and S-100B in group A at basal time were (2.51±0.67) μg/L, (21.53±3.84) ng/L respectively, remaining unchanged throughout the study. There were no differences between serum NSE and S-100B in groups B and C from those in group A at the first three time points. One hour after reperfusion, serum NSE and S-100B increased, then reached their peaks 6 h after reperfusion. Serum NSE and S-100B in group B were much higher than those in group C 3 h, 6 h after reperfusion. There was significant positive correlation between serum NSE and S-100B ( r =0.736, P < 0.001). Changes in serum TNF-α, IL-1, IL-8 2 Details are shown in Fig. 2 2 . There were no differences for serum TNF-α, IL-1, IL-8 between the three groups at the basal point. Compared with that at basal time point, TNF-α in group B and C increased significantly 30 min after ischemia (0 h) and reached its peak 3 h after reperfusion; these were correspondingly much higher than those in group A; TNF-α at time points of 1 h, 3 h in group B were significantly higher than that in group C, while there were no difference at other time points. IL-1 and IL-8 in groups B and C increased significantly 0.5 h after reperfusion and reached their peaks 3 h, 6 h after reperfusion respectively, and were much higher than those in group A; IL-1 at time points of 3 h, 6 h in group B were significantly higher than those in group C; IL-8 at time points of 1 h, 3 h in group B were significantly higher than those in group C. Brain water content 2 Brain water content was significantly higher in groups B and C (80.19%±0.70%, 80.45%±0.30%) as compared with 78.49%±0.59% in group A ( F =6.519, P =0.006; q =3.610, 4.933 and P =0.047, 0.006 respectively). But there was no difference between groups B and C ( q =1.322, P =0.625). Cerebral histopathological damages 2 As shown in Fig. 3 3 , neurons in group A had normal sizes, clear contour and axons, intact organelles in group B, neurons were shrunken and deeply stained, had unclear structures of organelles and significantly increased gap around neurons and microvessels. Damages of cerebral microscopic structures in group C were less severe than those in group B. Neurons had nearly normal sizes, slightly blurry contour and organelles, had less increased gap around neurons and microvessels. DISCUSSION 1 Complete cerebral ischemia could not be achieved by simplex occlusion of two common carotids and two vertebral arteries in the animals (Furlow, 1982 1982 ). When this combined with systemic hypotension induced by exsanguinations and reperfusion, changes of blood flow in the entire organism and brain following cardiac arrest and resuscitation are well simulated. So it may be accepted as an ideal animal model for cardiopulmonary cerebral resuscitation. Compared with the lower mean arterial blood pressure (35~40 mmHg) induced during ischemia in one published study (Wang et al., 2001 2001 ), about 50%~60% of basal values (50~60 mmHg) was maintained in the present experiment. In a preliminary study, we found that animals could hardly survive for more than 3 h with such low blood pressure during ischemia. Furthermore the rate and volume of exsanguination had to be well controlled to ensure a stable animal model. Neuron-specific enolase (NSE) is an isoenzyme of the glycplytic enzyme enolase (2-phospho-D-glycerate hydrolase), a dimeric enzyme composed of 2 gamma-subunits with a molecular weight of 78 kDa and biological half-life of approximately 24 h (Tiainen et al., 2003 2003 ). It is nearly exclusively located in neurons, neuroendocrine cells (APUD cells) and neuroendocrine tumors, including small-cell lung cancer (Tiainen et al., 2003 2003 ). S-100B is a dimeric acidic calcium-binding protein with a molecular weight of 21 kDa and biological half-life of 0.5 h (Jonsson et al., 2000 2000 ), and is found especially in high concentrations in astroglial cells that are prevalent in the white matter of the brain (Mussack et al., 2002 2002 ). When the brain suffers from different insults leading to impaired cellular structure and blood-brain-barrier integrity, NSE and S-100B are released quite rapidly into the cerebrospinal fluid and then into the circulation. Increased serum levels are proportionally related to the extent of cellular brain injury (Martens et al., 1998 1998 ). So serum NSE and S-100B are sensitive and quantitative markers of parenchymal brain injury due to ischemia stroke, intracerebral hemorrhage, trauma and cardiopulmonary surgery (Mabe et al., 1991 1991 ; Skogseid et al., 1992 1992 ; Schaarschmidt et al., 1994 1994 ; Isgro et al., 1997 1997 ; Missler et al., 1997 1997 ; Elting et al., 2000 2000 ; Jonsson et al., 2001 2001 ). Cardiac arrest leads to a period of temporary systemic and global cerebral ischemia, while the following cardiopulmonary resuscitation causes reperfusion injury, mediated by a series of inflammatory responses (Jean et al., 1998 1998 ). As a consequence, the observed significantly elevated NSE and S-100B levels in the blood served as predictors of neurological outcome in several previous studies (Martens et al., 1998 1998 ; Schoerkhuber et al., 1999 1999 ; Rosen et al., 2001 2001 ; Mussack et al., 2002 2002 ; Tiainen et al., 2003 2003 ). In the present study, serum NSE and S-100B began to increase 1 h after reperfusion, continued to rise and reached their peaks 6 h later in groups B and C. The good correlation between the two indices ( r =0.736) indicated increases of serum NSE and S-100B occurred very early after global cerebral ischemia-reperfusion injury. In group C treated with basic fibroblast growth factor, changes of blood pressure, heart rate and body temperature were similar to those in group B. But serum NSE and S-100B levels were lower at corresponding time points in group C, and with less severe cerebral histopathological damages. So it could be deduced that bFGF alleviated global cerebral ischemia-reperfusion injury, which was in accordance with previous findings showing that bFGF protected the brain from different insults. Himmelseher et al.( 1997 1997 ) found that bFGF increased cell survival and supported axonal re-elongations in adult hippocampal neurons in vitro when applied after axotomy. In cats undergoing left middle cerebral artery (MCA) occlusion for 4 h, intravenous bFGF at a rate of 5, 50, 250 μg/kg per hour decreased injury volume of ipsilateral cerebral cortex (Bethel et al., 1997 1997 ). A continuous intravenous infusion of 45 μg/(kg·h) bFGF for 3 h, beginning 30 min after fluid-percussion brain injury, reduced both the number of damaged neurons and the contusion volume in rats (Dietrich et al., 1996 1996 ). Brain water content was not decreased by bFGF in the present study, with results different from those of Liu and Zhu ( 1998 1998 ) who found that bFGF minimized brain water content in rat following forebrain ischemia-reperfusion, which could probably be attributable to different animal models employed. In this study, the test animals underwent complete global cerebral ischemia and longer time of reperfusion, which led to much severer brain injury. Inflammatory response plays a main causative role in cerebral ischemia-reperfusion injury, in which a variety of inflammatory mediators are involved. Increased levels of TNF-α are observed early in cerebral ischemia and reperfusion (Liu et al., 1994 1994 ; Buttini et al., 1996 1996 ). The sources of TNF-α under these conditions include activated macrophages/monocytes, neurons, astrocytes and microglias. TNF-α recruits and activates leukocytes, leading to increased leukocyte-endothelial adhesion (Fabry et al., 1992 1992 ; Liu et al., 1994 1994 ), which progressively induces expression of other cytokines, including IL-1, IL-6, IL-8 and so on, thus forming a positive feedback loop further exacerbating brain injury. Levels of TNF-α, IL-1 and IL-8 are positively correlated proportionably to neurons damage, brain edema and infarction extension, and can serve as good indicators of cerebral ischemia-reperfusion injury (Yamasaki et al., 1995 1995 ; Barone et al., 1997 1997 ; Yang et al., 1997 1997 ). Elevated serum IL-8 and TNF-α levels comprise the worse outcome in patients following out-of-hospital cardiac arrest (Ito et al., 2001 2001 ). Blocking or countering their activities lead to decreased neutrophil infiltration, blood-brain barrier breakdown, infarct size and extent of injury (Kadoya et al., 1995 1995 ; Matsumoto et al., 1997 1997 ; Yamasaki et al., 1997 1997 ; Yang et al., 1997 1997 ). In the present study, serum TNF-α was increased 30 min after ischemia, continued to rise after reperfusion, concomitant with increased serum IL-1 and IL-8. In those treated with bFGF, serum TNF-α, IL-1 and IL-8 were significantly decreased 1, 3, 6 h after reperfusion, compared with those in group B. It may indicate that bFGF probably exerts its neuroprotective effect by interfering with the expression and functions of these cytokines. In rats undergoing superior mesenteric artery occlusion (45 min) and reperfusion (3 d), an intravenous administration of 4 μg bFGF at the onset of reperfusion reduced serum tumor necrosis factor-alpha and free radicals, while protecting gut and liver against ischemia and reperfusion injury (Fu et al., 1997 1997 ). bFGF could suppress apoptosis of L929 cells induced by TNF-α, which was mediated by activation of the MAPK (mitogen activated protein kinases) intracellular signaling pathways (Gardner and Johnson, 1996 1996 ). bFGF also reduced IL-1 secretion (Bodo et al., 2000 2000 ), and down-regulated expression of vascular cell adhesion molecules stimulated by TNF-α and IL-1, resulting in markedly decreased levels of leukocyte recruitment and adhesion (Kikuchi, 2000 2000 ; Tromp et al., 2000 2000 ), subsequently decreasing tissue injury. CONCLUSION 1 The present study suggests that basic fibroblast growth factor alleviates brain injury following global ischemia reperfusion. And this is probably achieved by down-regulating expression of inflammatory factors and countering their activities to reduce inflammatory injury during reperfusion. It shows promising application in patients with cardiac arrest and deserves further investigations. References FC Barone , B Arvin , RF White , A Miller , CL Webb , RN Willette , PG Lysko , GZ Feuerstein . Tumor necrosis factor-alpha. A mediator of focal ischemic brain injury . Stroke , 1997 . 28 ( 6 ): 1233 - 1244 . . Tumor necrosis factor-alpha. A mediator of focal ischemic brain injury A Bethel , JR Kirsch , RC Koehler , SP Finklestein , RJ Traystman . Intravenous basic fibroblast growth factor decreases brain injury resulting from focal ischemia in cats . Stroke , 1997 . 28 ( 3 ): 609 - 615 . . Intravenous basic fibroblast growth factor decreases brain injury resulting from focal ischemia in cats M Bodo , T Baroni , F Carinci , E Becchetti , C Conte , C Bellucci , F Pezzetti , M Calvitti , S Bellocchio , G Stabellini , 等 . Interleukin secretion, proteoglycan and procollagen alpha (1) gene expression in Crouzon fibroblasts treated with basic fibroblast growth factor . Cytokine , 2000 . 12 ( 8 ): 1280 - 1283 . . Interleukin secretion, proteoglycan and procollagen alpha (1) gene expression in Crouzon fibroblasts treated with basic fibroblast growth factor TJ Bunch , CP West , DL Packer , MS Panutich , RD White . Admission predictors of in-hospital mortality and subsequent long-term outcome in survivors of ventricular fibrillation out-of-hospital cardiac arrest: A population-based study . Cardiology , 2004 . 102 ( 1 ): 41 - 47 . . Admission predictors of in-hospital mortality and subsequent long-term outcome in survivors of ventricular fibrillation out-of-hospital cardiac arrest: A population-based study M Buttini , K Appel , A Sauter , PJ Gebicke-Haerter , HW Boddeke . Expression of tumor necrosis factor alpha after focal cerebral ischemia in the rat . Neuroscience , 1996 . 71 ( 1 ): 1 - 16 . . Expression of tumor necrosis factor alpha after focal cerebral ischemia in the rat WD Dietrich , O Alonso , R Busto , SP Finklestein . Posttreatment with intravenous basic fibroblast growth factor reduces histopathological damage following fluid-percussion brain injury in rats . J Neurotrauma , 1996 . 13 ( 6 ): 309 - 316 . . Posttreatment with intravenous basic fibroblast growth factor reduces histopathological damage following fluid-percussion brain injury in rats JW Elting , AE de Jager , AW Teelken , MJ Schaaf , NM Maurits , J van der Naalt , CT Sibinga , GA Sulter , J de Keyser . Comparison of serum S-100 protein levels following stroke and traumatic brain injury . J Neuro Sci , 2000 . 181 ( 1-2 ): 104 - 110 . . Comparison of serum S-100 protein levels following stroke and traumatic brain injury Z Fabry , MM Waldschmidt , D Hendrickson , J Keiner , L Love-Homan , F Takei , MN Hart . Adhesion molecules on murine brain microvascular endothelial cells: Expression and regulation of ICAM-1 and Lgp 55 . J Neuroimmunol , 1992 . 36 ( 1 ): 1 - 11 . . Adhesion molecules on murine brain microvascular endothelial cells: Expression and regulation of ICAM-1 and Lgp 55 X Fu , Z Sheng , Y Wang , Y Ye , M Xu , T Sun , B Zhou . Basic fibroblast growth factor reduces the gut and liver morphologic and functional injuries after ischemia and reperfusion . J Trauma , 1997 . 42 ( 6 ): 1080 - 1085 . . Basic fibroblast growth factor reduces the gut and liver morphologic and functional injuries after ischemia and reperfusion TW Furlow . Cerebral ischemia produced by four-vessel occlusion in the rat: A quantitative evaluation of cerebral blood flow . Stroke , 1982 . 13 ( 6 ): 852 - 855 . . Cerebral ischemia produced by four-vessel occlusion in the rat: A quantitative evaluation of cerebral blood flow AM Gardner , GL Johnson . Fibroblast growth factor-2 suppression of tumor necrosis factor alpha-mediated apoptosis requires Ras and the activation of mitogen-activated protein kinase . J Biol Chem , 1996 . 271 ( 24 ): 14560 - 14566 . . Fibroblast growth factor-2 suppression of tumor necrosis factor alpha-mediated apoptosis requires Ras and the activation of mitogen-activated protein kinase S Himmelseher , E Pfenninger , M Georgieff . Effects of basic fibroblast growth factor on hippocampal neurons after axonal injury . J Trauma , 1997 . 42 ( 4 ): 659 - 664 . . Effects of basic fibroblast growth factor on hippocampal neurons after axonal injury F Isgro , C Schmidt , P Pohl , W Saggau . A predictive parameter in patients with brain related complications after cardiac surgery? . Eur J Cardiothorac Surg , 1997 . 11 ( 4 ): 640 - 644 . . A predictive parameter in patients with brain related complications after cardiac surgery? T Ito , D Saitoh , K Fukuzuka , T Kiyozumi , M Kawakami , T Sakamoto , Y Okada . Significance of elevated serum interleukin-8 in patients resuscitated after cardiopulmonary arrest . Resuscitation , 2001 . 51 ( 1 ): 47 - 53 . . Significance of elevated serum interleukin-8 in patients resuscitated after cardiopulmonary arrest WC Jean , SR Spellman , ES Nussbaum , WC Low . Reperfusion injury after focal cerebral ischemia: The role of inflammation and the therapeutic horizon . Neurosurgery , 1998 . 43 ( 6 ): 1382 - 1396 . . Reperfusion injury after focal cerebral ischemia: The role of inflammation and the therapeutic horizon H Jonsson , P Johnsson , P Hoglund , C Alling , S Blomquist . Elimination of S100B and renal function after cardiac surgery . J Cardiothorac Vasc Anesth , 2000 . 14 ( 6 ): 698 - 701 . . Elimination of S100B and renal function after cardiac surgery H Jonsson , P Johnsson , M Birch-Iensen , C Alling , S Westaby , S Blomquist . S100B as a predictor of size and outcome of stroke after cardiac surgery . Ann Thorac Surg , 2001 . 71 ( 5 ): 1433 - 1437 . . S100B as a predictor of size and outcome of stroke after cardiac surgery C Kadoya , EF Domino , GY Yang , JD Stern , AL Betz . Preischemic but not postischemic zinc protoporphyrin treatment reduces infarct size and edema accumulation after temporary focal cerebral ischemia in rats . Stroke , 1995 . 26 ( 6 ): 1035 - 1038 . . Preischemic but not postischemic zinc protoporphyrin treatment reduces infarct size and edema accumulation after temporary focal cerebral ischemia in rats H Kikuchi . Differential influences of bFGF and VEGF on the expression of vascular cell adhesion molecule-1 on human umbilical vein endothelial cells . Nihon Rinsho Meneki Gakkai Kaishi , 2000 . 23 ( 1 ): 12 - 21 . . Differential influences of bFGF and VEGF on the expression of vascular cell adhesion molecule-1 on human umbilical vein endothelial cells X Liu , XZ Zhu . Basic fibroblast growth factor protected forebrain against ischemia-reperfusion damage in rats . Acta Pharmacologica Sinica , 1998 . 19 ( 6 ): 527 - 530 . (in Chinese) . Basic fibroblast growth factor protected forebrain against ischemia-reperfusion damage in rats T Liu , RK Clark , PC McDonnell , PR Young , RF White , FC Barone , GZ Feuerstein . Tumor necrosis factor-alpha expression in ischemic neurons . Stroke , 1994 . 25 ( 7 ): 1481 - 1488 . . Tumor necrosis factor-alpha expression in ischemic neurons H Mabe , S Suzuki , M Mase , A Umemura , H Nagai . Serum neuron-specific enolase levels after subarachnoid hemorrhage . Surg Neurol , 1991 . 36 ( 3 ): 170 - 174 . . Serum neuron-specific enolase levels after subarachnoid hemorrhage P Martens , A Raabe , P Johnsson . Serum S-100 and neuron-specific enolase for prediction of regaining consciousness after global cerebral ischemia . Stroke , 1998 . 29 ( 11 ): 2363 - 2366 . . Serum S-100 and neuron-specific enolase for prediction of regaining consciousness after global cerebral ischemia T Matsumoto , K Ikeda , N Mukaida , A Harada , Y Matsumoto , J Yamashita , K Matsushima . Prevention of cerebral edema and infarct in cerebral reperfusion injury by an antibody to interleukin-8 . Lab Invest , 1997 . 77 ( 2 ): 119 - 125 . . Prevention of cerebral edema and infarct in cerebral reperfusion injury by an antibody to interleukin-8 U Missler , M Wiesmann , C Friedrich , M Kaps . S-100 protein and neuron-specific enolase concentrations in blood as indicators of infarction volume and prognosis in acute ischemic stroke . Stroke , 1997 . 28 ( 10 ): 1956 - 1960 . . S-100 protein and neuron-specific enolase concentrations in blood as indicators of infarction volume and prognosis in acute ischemic stroke T Mussack , P Biberthaler , KG Kanz , E Wiedemann , C Gippner-Steppert , W Mutschler , M Jochum . Serum S-100B and interleukin-8 as predictive markers for comparative neurologic outcome analysis of patients after cardiac arrest and severe traumatic brain injury . Crit Care Med , 2002 . 30 ( 12 ): 2669 - 2674 . . Serum S-100B and interleukin-8 as predictive markers for comparative neurologic outcome analysis of patients after cardiac arrest and severe traumatic brain injury M Pomfy , J Franko . Validation of a four-vessel occlusion model for transient global cerebral ischemia in dogs . J Hirnforsch , 1999 . 39 ( 4 ): 465 - 471 . . Validation of a four-vessel occlusion model for transient global cerebral ischemia in dogs H Rosen , KS Sunnerhagen , J Herlitz , C Blomstrand , L Rosengren . Serum levels of the brain-derived proteins S-100 and NSE predict long-term outcome after cardiac arrest . Resuscitation , 2001 . 49 ( 2 ): 183 - 191 . . Serum levels of the brain-derived proteins S-100 and NSE predict long-term outcome after cardiac arrest H Schaarschmidt , HW Prange , H Reiber . Neuron-specific enolase concentrations in blood as a prognostic parameter in cerebrovascular diseases . Stroke , 1994 . 25 ( 3 ): 558 - 565 . . Neuron-specific enolase concentrations in blood as a prognostic parameter in cerebrovascular diseases W Schoerkhuber , H Kittler , F Sterz , W Behringer , M Holzer , M Frossard , S Spitzauer , AN Laggner . Time course of serum neuron-specific enolase. A predictor of neurological outcome in patients resuscitated from cardiac arrest . Stroke , 1999 . 30 ( 8 ): 1598 - 1603 . . Time course of serum neuron-specific enolase. A predictor of neurological outcome in patients resuscitated from cardiac arrest IM Skogseid , HK Nordby , P Urdal , E Paus , F Lilleaas . Increased serum creatine kinase BB and neuron specific enolase following head injury indicates brain damage . Acta Neurochir (Wien) , 1992 . 115 ( 3-4 ): 106 - 111 . . Increased serum creatine kinase BB and neuron specific enolase following head injury indicates brain damage SA Snyder-Ramos , BW Bottiger . Molecular markers of brain damage–clinical and ethical implications with particular focus on cardiac arrest . Restor Neurol Neurosci , 2003 . 21 ( 3-4 ): 123 - 139 . . Molecular markers of brain damage–clinical and ethical implications with particular focus on cardiac arrest M Tiainen , RO Roine , V Pettila , O Takkunen . Serum neuron-specific enolase and S-100B protein in cardiac arrest patients treated with hypothermia . Stroke , 2003 . 34 ( 12 ): 2881 - 2886 . . Serum neuron-specific enolase and S-100B protein in cardiac arrest patients treated with hypothermia SC Tromp , MG Oude Egbrink , RP Dings , S van Velzen , DW Slaaf , HF Hillen , GJ Tangelder , RS Reneman , AW Griffioen . Tumor angiogenesis factors reduce leukocyte adhesion in vivo . Int Immunol , 2000 . 12 ( 5 ): 671 - 676 . . Tumor angiogenesis factors reduce leukocyte adhesion in vivo XB Wang , X Kuang , HX Wu , N Non . Delayed protective effect of adenosine A1 receptors on brain following preconditioning ischemia in rabbits . Chin J Anaeth (Chin) , 2001 . 21 ( 1 ): 40 - 43 . . Delayed protective effect of adenosine A1 receptors on brain following preconditioning ischemia in rabbits Y Yamasaki , Y Matsuo , N Matsuura , H Onodera , Y Itoyama , K Kogure . Transient increase of cytokine-induced neutrophil chemoattractant, a member of the interleukin-8 family, in ischemic brain areas after focal ischemia in rats . Stroke , 1995 . 26 ( 2 ): 318 - 322 . . Transient increase of cytokine-induced neutrophil chemoattractant, a member of the interleukin-8 family, in ischemic brain areas after focal ischemia in rats Y Yamasaki , Y Matsuo , J Zagorski , N Matsuura , H Onodera , Y Itoyama , K Kogure . New therapeutic possibility of blocking cytokine-induced neutrophil chemoattractant on transient ischemic brain damage in rats . Brain Res , 1997 . 759 ( 1 ): 103 - 111 . . New therapeutic possibility of blocking cytokine-induced neutrophil chemoattractant on transient ischemic brain damage in rats GY Yang , YJ Zhao , BL Davidson , AL Betz . Overexpression of interleukin-1 receptor antagonist in the mouse brain reduces ischemic brain injury . Brain Res , 1997 . 751 ( 2 ): 181 - 188 . . Overexpression of interleukin-1 receptor antagonist in the mouse brain reduces ischemic brain injury S Yoshimura , T Teramoto , MJ Whalen , MC Irizarry , Y Takagi , J Qiu , J Harada , C Waeber , XO Breakefield , MA Moskowitz . FGF-2 regulates neurogenesis and degeneration in the dentate gyrus after traumatic brain injury in mice . J Clin Invest , 2003 . 112 ( 8 ): 1202 - 1210 . . FGF-2 regulates neurogenesis and degeneration in the dentate gyrus after traumatic brain injury in mice 10.1631/jzus.2005.B0637 2004-09-17 2004-11-05 2005-06-23 2022-01-05 Copyright © 2005, Journal of Zhejiang University Science 2005 Journal of Zhejiang University-SCIENCE B (Biomedicine and Biotechnology) 07 6B