Traumatic brain injury (TBI) is the leading cause of mortality and morbidity in children and is characterized by reduced aerobic cerebral energy metabolism early after injury, possibly due to impaired activity of the pyruvate dehydrogenase complex. Exogenous acetyl-L-carnitine (ALCAR) is metabolized in the brain to acetyl coenzyme A and subsequently enters the tricarboxylic acid cycle. ALCAR administration is neuroprotective in animal models of cerebral ischemia and spinal cord injury, but has not been tested for TBI. This study tested the hypothesis that treatment with ALCAR during the first 24 h following TBI in immature rats improves neurologic outcome and reduces cortical lesion volume. Postnatal day 21–22 male rats were isoflurane anesthetized and used in a controlled cortical impact model of TBI to the left parietal cortex. At 1, 4, 12 and 23 h after injury, rats received ALCAR (100 mg/kg, intraperitoneally) or drug vehicle (normal saline). On days 3–7 after surgery, behavior was assessed using beam walking and novel object recognition tests. On day 7, rats were transcardially perfused and brains were harvested for histological assessment of cortical lesion volume, using stereology. Injured animals displayed a significant increase in foot slips compared to sham-operated rats (6 ± 1 SEM vs. 2 ± 0.2 on day 3 after trauma; n = 7; p < 0.05). The ALCAR-treated rats were not different from shams and had fewer foot slips compared to vehicle-treated animals (2 ± 0.4; n = 7; p< 0.05). The frequency of investigating a novel object for saline-treated TBI animals was reduced compared to shams (45 ± 5% vs. 65 ± 10%; n = 7; p < 0.05), whereas the frequency of investigation for TBI rats treated with ALCAR was not significantly different from that of shams but significantly higher than that of saline-treated TBI rats (68 ± 7; p < 0.05). The left parietal cortical lesion volume, expressed as a percentage of the volume of tissue in the right hemisphere, was significantly smaller in ALCAR-treated than in vehicle-treated TBI rats (14 ± 5% vs. 28 ± 6%; p < 0.05). We conclude that treatment with ALCAR during the first 24 h after TBI improves behavioral outcomes and reduces brain lesion volume in immature rats within the first 7 days after injury.

Keywords:
Sensorimotor function, Controlled cortical impact, Novel object recognition, Memory, Hippocampus, Acetyl-L-carnitine, Neuroprotection
1.
Xu Y, McArthur DL, Alger JR, Etchepare M, Hovda DA, Glenn TC, Huang S, Dinov I, Vespa PM: Early nonischemic oxidative metabolic dysfunction leads to chronic brain atrophy in traumatic brain injury. J Cereb Blood Flow Metab 2010;30:883–894.
2.
Robertson CL, Scafidi S, McKenna MC, Fiskum G: Mitochondrial mechanisms of cell death and neuroprotection in pediatric ischemic and traumatic brain injury. Exp Neurol 2009;218:371–380.
3.
Verweij BH, Amelink GJ, Muizelaar JP: Current concepts of cerebral oxygen transport and energy metabolism after severe traumatic brain injury. Prog Brain Res 2007;161:111–124.
4.
Scafidi S, O’Brien J, Hopkins I, Robertson C, Fiskum G, McKenna M: Delayed cerebral oxidative glucose metabolism after traumatic brain injury in young rats. J Neurochem 2009;109(suppl 1):189–197.
5.
Vaishnav RA, Singh IN, Miller DM, Hall ED: Lipid peroxidation-derived reactive aldehydes directly and differentially impair spinal cord and brain mitochondrial function. J Neurotrauma 2010;27:1311–1320.
6.
Patel SP, Sullivan PG, Lyttle TS, Rabchevsky AG: Acetyl-L-carnitine ameliorates mitochondrial dysfunction following contusion spinal cord injury. J Neurochem 2010;114:291–301.
7.
Sharma P, Benford B, Li ZZ, Ling GS: Role of pyruvate dehydrogenase complex in traumatic brain injury and measurement of pyruvate dehydrogenase enzyme by dipstick test. J Emerg Trauma Shock 2009;2:67–72.
8.
Opii WO, Nukala VN, Sultana R, Pandya JD, Merchant ML, Klein JB, Sullivan PG, Butterfield DA: Proteomic identification of oxidized mitochondrial proteins following experimental traumatic brain injury. J Neurotrauma 2007;24:772–789.
9.
Thomale UW, Griebenow M, Mautes A, Beyer TF, Dohse NK, Stroop R, Sakowitz OW, Unterberg AW: Heterogeneous regional and temporal energetic impairment following controlled cortical impact injury in rats. Neurol Res 2007;29:594–603.
10.
Robertson CL, Saraswati M, Fiskum G: Mitochondrial dysfunction early after traumatic brain injury in immature rats. J Neurochem 2007;101:1248–1257.
11.
Richards EM, Rosenthal RE, Kristian T, Fiskum G: Postischemic hyperoxia reduces hippocampal pyruvate dehydrogenase activity. Free Radical Biol Med 2006;40:1960–1970.
12.
Vereczki V, Martin E, Rosenthal RE, Hof PR, Hoffman GE, Fiskum G: Normoxic resuscitation after cardiac arrest protects against hippocampal oxidative stress, metabolic dysfunction, and neuronal death. J Cereb Blood Flow Metab 2005;26:821–835.
13.
Bogaert YE, Rosenthal RE, Fiskum G: Postischemic inhibition of cerebral cortex pyruvate dehydrogenase. Free Radic Biol Med 1994;16:811–820.
14.
Bogaert YE, Sheu KF, Hof PR, Brown AM, Blass JP, Rosenthal RE, Fiskum G: Neuronal subclass-selective loss of pyruvate dehydrogenase immunoreactivity following canine cardiac arrest and resuscitation. Exp Neurol 2000;161:115–126.
15.
Xing G, Ren M, Watson WA, O’Neil JT, Verma A: Traumatic brain injury-induced expression and phosphorylation of pyruvate dehydrogenase: a mechanism of dysregulated glucose metabolism. Neurosci Lett 2009;454:38–42.
16.
Prins ML: Cerebral metabolic adaptation and ketone metabolism after brain injury. J Cereb Blood Flow Metab 2008;28:1–16.
17.
Faria MH, Muniz LR, Vasconcelos PR: Ketone bodies metabolism during ischemic and reperfusion brain injuries following bilateral occlusion of common carotid arteries in rats. Acta Cir Bras 2007;22:125–129.
18.
Koppaka SS, Puchowicz, LaManna JC, Gatica JE: Effect of alternate energy substrates on mammalian brain metabolism during ischemic events. Adv Exp Med Biol 2008;614:361–370.
19.
Dardzinski BJ, Smith SL, Towfighi J, Williams GD, Vannucci RC, Smith MB: Increased plasma β-hydroxybutyrate, preserved cerebral energy metabolism, and amelioration of brain damage during neonatal hypoxia ischemia with dexamethasone pretreatment. Pediatr Res 2000;48:248–255.
20.
Massieu L, Haces ML, Montiel T, Hernandez-Fonseca K: Acetoacetate protects hippocampal neurons against glutamate-mediated neuronal damage during glycolysis inhibition. Neuroscience 2003;120:365–378.
21.
Rosenthal RE, Williams R, Bogaert YE, Getson PR, Fiskum G: Prevention of postischemic canine neurological injury through potentiation of brain energy metabolism by acetyl-L-carnitine. Stroke 1992;23:1312–1317.
22.
Zanelli SA, Solenski NJ, Rosenthal RE, Fiskum G: Mechanisms of ischemic neuroprotection by acetyl-L-carnitine. Ann NY Acad Sci 2005;1053:153–161.
23.
Ames BN, Liu J: Delaying the mitochondrial decay of aging with acetylcarnitine. Ann NY Acad Sci 2004;1033:108–116.
24.
Jones LL, McDonald DA, Borum PR: Acylcarnitines: role in brain. Prog Lipid Res 2010;49:61–75.
25.
Virmani MA, Caso V, Spadoni A, Rossi S, Russo F, Gaetani F: The action of acetyl-L-carnitine on the neurotoxicity evoked by amyloid fragments and peroxide on primary rat cortical neurones. Ann NY Acad Sci 2001;939:162–178.
26.
Calabrese V, Mancuso C, Pennisi G, Calafato S, Bellia F, Guiffrida Stella AM, Schapira T, Dinkova Kostova AT, Rizzarelli E: Cellular stress response: a novel target for chemoprevention and nutritional neuroprotection in aging, neurodegenerative disorders and longevity. Neurochem Res 2008;33:2444–2471.
27.
Calabrese V, Ravagna A, Colombrita C, Scapagnini G, Guagliano E, Calvani M, Butterfield DA, Guiffrida Stella AM: Acetylcarnitine induces heme oxygenase in rat astrocytes and protects against oxidative stress: involvement of the transcription factor Nrf2. J Neurosci Res 2005;79:509–521.
28.
Fiskum G, Liu Y, Bogaert YE, Rosenthal RE: Acetyl-L-carnitine stimulates cerebral oxidative metabolism and inhibits protein oxidation following cardiac arrest in dogs; in Krieglstein J, Oberpichler-Schwenk H (eds): Pharmacology of Cerebral Ischemia. Stuttgart, Wissenschaftliche Verlagsgesellschaft, 1992, pp 487–491.
29.
Ishii T, Shimpo Y, Matsuoka Y, Kinoshita K: Anti-apoptotic effect of acetyl-L-carnitine and L-carnitine in primary cultured neurons. Jpn J Pharmacol 2000;83:119–124.
30.
Al-Majed AA, Sayed-Ahmed MM, Al-Omar FA, Al-Yahya AA, Aleisa AM, Al-Shabanah OA: Carnitine esters prevent oxidative stress damage and energy depletion following transient forebrain ischaemia in the rat hippocampus. Clin Exp Pharmacol Physiol 2006;33:725–733.
31.
Pettegrew JW, Panchalingam K, Withers G, McKeag D, Strychor S: Changes in brain energy and phospholipid metabolism during development and aging in the Fischer 344 rat. J Neuropathol Exp Neurol 1990;49:237–249.
32.
Shuaib A, Waqaar T, Wishart T, Kanthan R, Howlett W: Acetyl-L-carnitine attenuates neuronal damage in gerbils with transient forebrain ischemia only when given before the insult. Neurochem Res 1995;20:1021–1025.
33.
Lolic MM, Fiskum G, Rosenthal RE: Neuroprotective effects of acetyl-L-carnitine after stroke in rats. Ann Emerg Med 1997;29:758–765.
34.
Jalal FY, Bohlke M, Maher TJ: Acetyl-L-carnitine reduces the infarct size and striatal glutamate outflow following focal cerebral ischemia in rats. Ann NY Acad Sci 2010;1199:95–104.
35.
Hagberg H, Mallard C, Rousset CI, Wang X: Apoptotic mechanisms in the immature brain: involvement of mitochondria. J Child Neurol 2009;24:1141–1146.
36.
Bayir H, Kochanek PM, Kagan VE: Oxidative stress in immature brain after traumatic brain injury. Dev Neurosci 2006;28:420–431.
37.
Bauer R, Fritz H: Pathophysiology of traumatic injury in the developing brain: an introduction and short update. Exp Toxicol Pathol 2004;56:65–73.
38.
Scafidi S, Fiskum G, Lindauer SL, Bamford P, Shi D, Hopkins I, McKenna MC: Metabolism of acetyl-L-carnitine for energy and neurotransmitter synthesis in the immature rat brain. J Neurochem 2010;114:820–831.
39.
Carter RJ, Morton J, Dunnett SB: Motor coordination and balance in rodents. Curr Protoc Neurosci 2001;chapt 8:unit 8.12.
40.
Bevins RA, Besheer J: Object recognition in rats and mice: a one-trial non-matching-to-sample learning task to study ‘recognition memory’. Nat Protoc 2006;1:1306–1311.
41.
Ennaceur A: One-trial object recognition in rats and mice: methodological and theoretical issues. Behav Brain Res 2010;215:244–254.
42.
Ennaceur A, Delacour J: A new one-trial test for neurobiological studies of memory in rats. 1. Behavioral data. Behav Brain Res 1988;31:47–59.
43.
Hall ED, Bryant YD, Cho W, Sullivan PG: Evolution of post-traumatic neurodegeneration after controlled cortical impact traumatic brain injury in mice and rats as assessed by the de Olmos silver and fluorojade staining methods. J Neurotrauma 2008;25:235–247.
44.
Reger ML, Hovda DA, Giza CC: Ontogeny of rat recognition memory measured by the novel object recognition task. Dev Psychobiol 2009;51:672–678.
45.
Broadbent NJ, Squire LR, Clark RE: Spatial memory, recognition memory, and the hippocampus. Proc Natl Acad Sci USA 2004;101:14515–14520.
46.
Aureli T, Miccheli A, di Cocco ME, Ghirardi O, Giuliani A, Ramacci MT, Conti F: Effect of acetyl-L-carnitine on recovery of brain phosphorus metabolites and lactic acid level during reperfusion after cerebral ischemia in the rat: study by 13P- and 1H-NMR spectroscopy. Brain Res 1994;643:92–99.
47.
Parnetti L, Gaiti A, Mecocci P, Cadini D, Senin U: Pharmacokinetics of IV and oral acetyl-L-carnitine in a multiple dose regimen in patients with senile dementia of Alzheimer type. Eur J Clin Pharmacol 1992;42:89–93.
48.
Youle M, Osio M: A double-blind, parallel-group, placebo-controlled, multicentre study of acetyl-L-carnitine in the symptomatic treatment of antiretroviral toxic neuropathy in patients with HIV-1 infection. HIV Med 2007;8:241–250.
49.
Appelberg KS, Hovda DA, Prins ML: The effects of a ketogenic diet on behavioral outcome after controlled cortical impact injury in the juvenile and adult rat. J Neurotrauma 2009;26:497–506.
50.
Fox GB, Faden AI: Traumatic brain injury causes delayed motor and cognitive impairment in a mutant mouse strain known to exhibit delayed Wallerian degeneration. J Neurosci Res 1998;53:718–727.
51.
Fox GB, Fan L, LeVasseur RA, Faden AI: Effect of traumatic brain injury on mouse spatial and nonspatial learning in the Barnes circular maze. J Neurotrauma 1998;15:1037–1046.
52.
Boyeson MG, Callister TR, Cavazos JE: Biochemical and behavioral effects of a sensorimotor cortex injury in rats pretreated with the noradrenergic neurotoxin DSP-4. Behav Neurosci 1992;106:964–973.
53.
Allen GV, Chase T: Induction of heat shock proteins and motor function deficits after focal cerebellar injury. Neuroscience 2001;102:603–614.
54.
Williams AJ, Ling GS, Tortella FC: Severity level and injury track determine outcome following a penetrating ballistic-like brain injury in the rat. Neurosci Lett 2006;408:183–188.
55.
Squire LR, Wixted JT, Clark RE: Recognition memory and the medial temporal lobe: a new perspective. Nat Rev Neurosci 2007;8:872–883.
56.
Prins ML, Fujima LS, Hovda DA: Age-dependent reduction of cortical contusion volume by ketones after traumatic brain injury. J Neurosci Res 2005;82:413–420.
57.
Prins ML, Lee SM, Fujima LS, Hovda DA: Increased cerebral uptake and oxidation of exogenous BHB improves ATP following traumatic brain injury in adult rats. J Neurochem 2004;90:666–672.
58.
Winter BK, Fiskum G, Gallo LL: Effects of L-carnitine on serum triglyceride and cytokine levels in rat models of cachexia and septic shock. Br J Cancer 1995;72:1173–1179.
59.
Poon HF, Calabrese V, Calvani M, Butterfield DA: Proteomics analyses of specific protein oxidation and protein expression in aged rat brain and its modulation by L-acetylcarnitine: insights into the mechanisms of action of this proposed therapeutic agent for CNS disorders associated with oxidative stress. Antioxid Redox Signal 2006;8:381–394.
60.
Ferraresi R, Troiano L, Roat E, Nemes E, Lugli E, Nasi M, Pinti M, Calvani M, Iannucelli M, Cossarizza A: Protective effect of acetyl-L-carnitine against oxidative stress induced by antiretroviral drugs. FEBS Lett 2006;580:6612–6616.
61.
Shen W, Liu K, Tian C, Yang L, Li X, Ren J, Packer L, Head E, Sharman E, Liu J: Protective effects of R-α-lipoic acid and acetyl-L-carnitine in MIN6 and isolated rat islet cells chronically exposed to oleic acid. J Cell Biochem 2008;104:1232–1243.
62.
Liu Y, Rosenthal RE, Starke-Reed P, Fiskum G: Inhibition of postcardiac arrest brain protein oxidation by acetyl-L-carnitine. Free Radic Biol Med 1993;15:667–670.
63.
Liu J, Head E, Kuratsune H, Cotman CW, Ames BN: Comparison of the effects of L-carnitine and acetyl-L-carnitine on carnitine levels, ambulatory activity, and oxidative stress biomarkers in the brain of old rats. Ann NY Acad Sci 2004;1033:117–131.
64.
Loots DT, Mienie LJ, Bergh JJ, van der Schyf CJ: Acetyl-L-carnitine prevents total body hydroxyl free radical and uric acid production induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in the rat. Life Sci 2004;75:1243–1253.
65.
Yasui F, Matsugo S, Ishiashi M, Kajita T, Ezashi Y, Oomura Y, Kojo S, Sasaki K: Effects of chronic acetyl-L-carnitine treatment on brain lipid hydroperoxide level and passive avoidance learning in senescence-accelerated mice. Neurosci Lett 2002;334:177–180.
66.
Barhwal K, Hota SK, Jain V, Prasad D, Singh SB, Ilavazhagan G: Acetyl-L-carnitine (ALCAR) prevents hypobaric hypoxia-induced spatial memory impairment through extracellular related kinase-mediated nuclear factor erythroid 2-related factor 2 phosphorylation. Neuroscience 2009;161:501–514.
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2011
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