Nimodipine does not affect cerebral lactate levels following complete ischemia in dogs

Nimodipine is known to improve postischemic cerebral blood flow (CBF) and neurologic outcome in experimental animals. Whether or not the two observations are related is unknown. This study searched for a possible improved rate of brain metabolic recovery in animals treated with nimodipine postischemia. Complete cerebral ischemia was produced for 11 min in 16 dogs, followed by reperfusion for 70 min. Prior to ischemia, glucose was administered (0.75 g X kg-1) in 12 dogs. Half of the glucose-treated dogs were given i.v. nimodipine, beginning 5 min postischemia (10 micrograms X kg-1 bolus followed by 1 microgram X kg-1 X min-1). The other half were given only saline postischemia. The remaining four dogs were given no glucose and received saline only postischemia. In all dogs, serial brain biopsies were taken at 2, 20, 40, and 70 min postischemia. In 5 dogs, the integrity of the blood-brain barrier (BBB) was tested by injection of Evans blue dye and postmortem examination of the brains. Brain biopsies were assayed for concentrations of phosphocreatine, ATP, ADP, AMP, glucose, lactate, and pyruvate. In all dogs, there was rapid restoration of a normal brain energy state following reperfusion. Brain lactate had returned to near normal in all dogs by 70 min postischemia, and the rate of lactate depletion was not different between groups. The integrity of the BBB was only minimally affected. A portion of the brain lactate was converted to pyruvate rather than crossing the BBB.(ABSTRACT TRUNCATED AT 250 WORDS)     

Nicardipine increases cerebral blood flow but does not improve neurologic recovery in a canine model of complete cerebral ischemia

The effects of the calcium entry blocker nicardipine on CBF, CMRO2, and neurologic outcome following 10 min of complete cerebral ischemia were examined in dogs. In CBF and CMRO2 studies, the CBF in the untreated group (seven dogs) and the nicardipine group (seven dogs; 20 micrograms kg-1 at 30 min postischemia and a subsequent infusion of 2 micrograms kg-1 min-1 for 90 min) initially increased to 300-400% and then returned to preischemic values at 30 min postischemia. Thereafter the CBF in the untreated group significantly decreased to 50% of preischemic values for the following 90-min period (hypoperfusion), while the CBF in the nicardipine group did not differ from preischemic values. The CMRO2 in both groups decreased to approximately 50-80% of preischemic values after 15 min postischemia and did not differ between the groups throughout the study. In neurologic outcome studies, 18 dogs were divided into three groups (of six dogs each): untreated; saline infusion only, posttreated; nicardipine as in CBF and CMRO2 studies, pretreated; nicardipine 20 micrograms kg-1 at 2 min preischemia and a subsequent infusion of 2 micrograms kg-1 min-1 from immediately postischemia to 120 min postischemia. Nicardipine treatment initiated either before or after ischemia failed to improve neurologic outcome at 48 h postischemia. Thus, the increase of postischemic global CBF by nicardipine is not accompanied by neurologic recovery in a canine model of complete cerebral ischemia.     

Tirilazad reduces cortical infarction after transient but not permanent focal cerebral ischemia in rats.

BACKGROUND AND PURPOSE: We examined the cytoprotective effect of the lipid peroxidation inhibitor tirilazad mesylate (U74006F) in rodent models of neocortical infarction induced by transient and permanent focal cerebral ischemia. METHODS: Wistar rats (experiment 1) and spontaneously hypertensive rats (experiment 2) were subjected to 2 hours of transient middle cerebral artery occlusion followed by 22 hours of reperfusion and pretreated with 10 mg/kg i.p. tirilazad, vehicle, or saline. Repeat doses were given at 4 and 10 hours after reperfusion. Spontaneously hypertensive rats were also subjected to permanent middle cerebral artery occlusion and either pretreated with tirilazad, vehicle, or saline intraperitoneally (experiment 3) or treated with either tirilazad or vehicle intravenously after ischemia (experiment 4). Cortical infarct volumes were measured 24 hours after the onset of either transient or permanent ischemia, and changes in core regional cerebral blood flow were monitored with laser Doppler flowmetry. RESULTS: Tirilazad reduced infarct volume after transient ischemia by 40% in Wistar rats (p = 0.08) (experiment 1) and 23% in spontaneously hypertensive rats (p less than 0.05) (experiment 2) but did not reduce infarction after permanent ischemia whether it was given intraperitoneally (experiment 3) or intravenously (experiment 4). Ischemic core blood flows were not affected during ischemia, nor were they affected during reperfusion after transient ischemia. CONCLUSIONS: Tirilazad reduces cortical infarction in transient but not permanent ischemia, an effect not related to improvement in regional cerebral blood flow. Tirilazad might prove to be useful as an adjuvant therapy after successful thrombolysis in acute stroke patients.     

Allopurinol pretreatment improves evoked response recovery following global cerebral ischemia in dogs

The reperfusion of previously ischemic tissue may lead to the formation of highly reactive free radicals that promote tissue injury. Xanthine oxidase has been implicated as one source of these free radicals. We examined the role of xanthine oxidase in brain injury using a cerebrospinal fluid compression model of global cerebral ischemia with 15 minutes of ischemia and 4 hours of reperfusion. Seven dogs were pretreated with the xanthine oxidase inhibitor allopurinol (50 mg/kg for 5 days). Neurophysiological recovery was monitored with cortical somatosensory evoked potentials. As an attempt to correlate brain recovery with the mechanism of protection, free brain malondialdehyde was measured at the end of reperfusion by high-performance liquid chromatography. Brain water content was measured by wet-dry weights. Compared with seven untreated control dogs, allopurinol pretreatment significantly improved recovery of somatosensory evoked potentials after 4 hours of reperfusion. However, the amount of free malondialdehyde in the allopurinol-treated dogs was 32% greater than that in the controls. Brain water content was similar in the two groups. These results suggest that xanthine oxidase contributes to brain injury after ischemia and reperfusion. However, tissue damage caused by xanthine oxidase may be mediated through mechanisms other than free radical production.     

Lidoflazine does not improve neurologic outcome when administered after complete cerebral ischemia in primates

In order to investigate the effects of the calcium entry blocker lidoflazine on neurologic outcome in primates following an episode of global brain ischemia, 12 pigtail monkeys (Macaca nemestrina) were subjected to 17 min of complete cerebral ischemia, followed by 48 h of intensive care treatment and daily neurologic evaluations for 96 h. The monkeys were randomly assigned to receive, in a blind fashion, either lidoflazine 1.0 mg/kg (n = 6) or inactive lidoflazine solvent (n = 6) at 5 min, 8 h, and 16 h postischemia. One monkey in the lidoflazine group did not meet preestablished protocol criteria and was excluded from data analysis. The remaining monkeys were well matched for age, sex, and other physiologic variables. Neurologic outcome was not significantly different between the lidoflazine- and placebo-treated groups (p greater than 0.5). No monkey in either group achieved a normal neurologic exam by 96 h postischemia. Three lidoflazine-treated monkeys and two placebo-treated monkeys died prior to the 96-h neurologic evaluation. These deaths were judged to be neurologic in origin. The authors concluded that lidoflazine does not improve neurologic outcome in primates when administered after 17 min of complete cerebral ischemia.     

Cerebral metabolic state following complete compression ischemia

Recovery of cerebral metabolism after total ischemia was studied in rats by measurements of brain tissue concentrations of carbohydrate substrates, amino acids and organic phosphates at various periods (1–180 min) following an increase in the intracranial CSF pressure to values exceeding the mean arterial blood pressure. Control experiments were performed to evaluate the recovery of the EEG and of gross neurological function.Following ischemia of up to 15 min duration there was rapid rephosphorylation of PCr and ATP suggesting that mitochondrial function was adequately resumed. Furthermore, since the AMP levels were completely normalized and since accumulated lactate disappeared the data exclude the possibility that any significant proportion of the tissue remained unperfused.After periods of ischemia lasting 5 min or more EEG or gross neurological function did not normalize within the period studied (180 min). Although the adenylate energy charge returned to normal after 3 min of ischemia significantly subnormal values were obtained after 7.5 and 15 min of ischemia, suggesting that a biochemical lesion may have occurred.Recirculation of the tissue after ischemia was followed by disappearance of accumulated ammonia and by replenishment of citric cycle intermediates. However, α-ketoglutarate and malate showed slightly subnormal values after 180 min. Furthermore, there was a permanent decrease in glutamate and a late increase in glutamine indicating a lingering derangement in the amino acid metabolism that many have functional implications.     

Ischemic preconditioning does not improve neurological recovery after spinal cord compression injury in the rat

Ischemic preconditioning (IPC) has been defined as the endogenous cellular protective mechanism evoked by brief ischemic periods. IPC renders the tissue of the central nervous system more resistant to subsequent lethal ischemic insults, and similar protective effect of IPC has been observed after experimental traumatic brain injury. Spinal cord trauma differs from cerebral trauma in that the secondary processes are damaging mostly the white matter. In the present study, we have tested the hypothesis that a transient non-lethal ischemic insult would improve outcomes after subsequent traumatic spinal cord injury (SCI). In the IPC group, 5-min spinal cord ischemia has been induced by aortic occlusion combined with hypotension. Forty-eight hours after IPC, moderate spinal cord injury has been induced by epidural balloon inflation at T8 level. Control group underwent identical surgical procedures without ischemia followed by SCI after 48 h. During the 4-week survival, locomotor performance of all rats was repeatedly tested and evaluated according to BBB scale. After 4 weeks, the animals were perfusion-fixed for histopathology, and morphometric analyses were performed in order to quantify the extent of the spinal cord lesion. All animals were completely paraplegic after SCI, and showed partial neurological recovery during their survival period. No significant differences were detected either in neurological scores or in morphometric measurements after 4 weeks' survival. These results indicate that in contrary to cerebral trauma, IPC does not improve the outcome after SCI.     

17beta-Estradiol treatment following permanent focal ischemia does not influence recovery of sensorimotor function

The development of therapy to aid poststroke recovery is essential. The female hormone 17beta-estradiol has been shown to promote synaptogenesis; the purpose of this study was to attempt to harness these mechanisms to promote repair and recovery in the peri-infarct zone. Rats were ovariectomized, tested for sensorimotor function, and the middle cerebral artery permanently occluded (MCAO). Infarct volumes were calculated using MRI, and damage was equivalent in all animals prior to implantation of either 17beta-estradiol or placebo pellets. Animals were tested for functional recovery for 28 days and tissue processed for synaptic marker syntaxin immunohistochemistry. The stroke induced a significant behavioral deficit, which persisted out to 28 days, and was not significantly different between 17beta-estradiol and placebo treatment groups. There was no difference in syntaxin immunostaining between groups in either the peri-infarct cortex or in the dendritic CA1 reference region. In conclusion, 17beta-estradiol treatment, delivered poststroke, did not influence recovery of function or synaptogenesis.     

Effect of nimodipine on cerebral functional and metabolic recovery following ischemia in the rat brain

Whether the calcium entry blocker, nimodipine, prevents the increase in the concentration of free fatty acids and metabolic disturbances during ischemia and promotes functional and metabolic recovery after recirculation were examined. Severe forebrain ischemia in rats was induced by four-vessel occlusion with mild hypotension. After 30 minutes of ischemia, recirculation was started by removal of the arterial clamps and by increasing blood pressure to the preischemic level. Recovery of EEG activity following recirculation was better in the nimodipine-treated group than in the control group. During the ischemic period, there were no significant differences in accumulation of free fatty acids or in depletion of ATP between treated and control groups. At 120 minutes following recirculation, recovery of the ATP level was significantly better in the treated group than in the control group. Therefore, the promotion of functional and metabolic recovery by nimodipine-treatment is suggested to be not due to the prevention of an accumulation of free fatty acids nor to the depletion of ATP during the ischemic period, but to either improvement of postischemic hypoperfusion or a direct action on metabolic processes during reperfusion period.     

Baclofen does not protect against cerebral ischemia in rats

Presynaptic release of glutamate into the extracellular compartment and activation of receptor-operated calcium channels may contribute to ischemic neuronal damage. We evaluated the effect of baclofen, a selective inhibitor of presynaptic glutamate release, on mortality, working memory, and light microscopic hippocampal and cortical damage in the four-vessel occlusion model of cerebral ischemia using 64 male Wistar rats. Baclofen (10 mg/kg i.p.) given 1 hour before and 30-60 minutes after 20 minutes of global ischemia did not lessen mortality, prevent ischemic cellular damage, or significantly improve working memory compared with no treatment. We conclude that preischemic and postischemic administration of baclofen does not protect neurons from ischemic injury.     

Forced exercise does not improve recovery after hemorrhagic stroke in rats

Exercise can improve recovery following ischemia and intracerebral hemorrhage (ICH) in rodents. We tested whether forced exercise (EX; running wheel) prior to and/or following ICH in rats would reduce lesion volume and improve functional outcome (walking, skilled reaching, spontaneous paw usage) at 7 weeks post-ICH. A striatal hemorrhage was produced by infusing collagenase. First, we compared animals that received EX (2 weeks; 1 h/day) ending two days prior to ICH and/or starting two weeks following ICH. EX did not improve functional recovery or affect lesion size. Doubling the amount of EX given per day (two 1-h sessions) both prior to and following ICH did not alter lesion volume, but worsened recovery. We then determined if EX (1 h/day) prior to and following ICH would affect outcome after a somewhat milder insult. There were no differences between the groups in lesion volume or recovery. Finally, we used a hemoglobin assay at 12 h following ICH to determine if pre-stroke EX (2 weeks; 1 h/day) aggravated bleeding. It did not. These observations suggest that EX does not improve outcome when given prior to and/or when delayed following ICH. Effective rehabilitation for ICH will likely require more complex interventions than forced running.     

Effect of calcium entry blocker, nimodipine, on the cerebral function and metabolic recovery following experimental cerebral ischemia

Ischemic deporalization of cell membranes is associated with a precipitous influx of calcium from the extracellular to the intracellular compartment, and it is suggested that increased intracellular calcium in ischemic brain leads to an activation of phospholipase and to increase of the concentration of free fatty acids, in particular arachidonic acid, with energy depletion. The objective of the present study is to test whether calcium entry blocker, nimodipine, prevent increase of free fatty acids and metabolic disturbances during ischemic period, and promote functional and metabolic recovery after recirculation. Severe forebrain ischemia in rats was induced by four-vessel occlusion with reducing the systolic arterial pressure to 100 mmHg. After forebrain ischemia had been maintained for 30 minutes, recirculation was started by removal of the arterial clamps of bilateral common carotid arteries and by increasing systemic blood pressure to the preischemic level. The EEG was continuously recorded from gold-coated screws inserted bilaterally in the parietal bones with the tips in extradural position, against a reference inserted prefrontal bone. Analysis of power spectrum of EEG activity was done by Berg Fourier Analyser. The brain were frozen in situ with liquid nitrogen before, during and after ischemia and then chiselled out during irrigation with liquid nitrogen. Concentrations of ATP, ADP, AMP and free fatty acids in brain tissue were determined with high performance liquid chromatography. Nimodipine, 10 micrograms/kg, was given intravenously 2-3 minutes before induction of ischemia, and an infusion of 1 microgram/kg/min was continued during ischemic and postischemic periods.(ABSTRACT TRUNCATED AT 250 WORDS)     

Failure of deferoxamine, an iron chelator, to improve neurologic outcome following complete cerebral ischemia in dogs

Eleven minutes of complete cerebral ischemia was produced in 17 dogs by temporary ligation of the venae cavae and aorta. Immediately prior to the ischemic episode, 7 dogs received deferoxamine, an iron chelator, 50 mg/kg i.v., and 10 dogs received an equivalent volume of saline placebo i.v. Five dogs failed to meet preestablished protocol criteria and were excluded from data analysis. Neurologic recovery was evaluated by an observer blind to the treatment groups in the remaining 12 dogs at 48 hours postischemia. The neurologic effects of complete cerebral ischemia were compared between dogs treated with deferoxamine and those receiving placebo treatment. One of 6 deferoxamine-treated dogs was normal and 5 were moderately to severely damaged. Similarly, 1 of 6 placebo-treated dogs was normal and 5 were moderately to severely damaged. The authors conclude that deferoxamine does not provide cerebral protection in this model of complete cerebral ischemia.     

The antioxidant tirilazad does not affect cerebral blood flow or histopathologic changes following severe ischemia in rats

We studied the effect of tirilazad, an aminosteroid with radical scavenging effect, or its vehicle on cerebral blood flow and neuronal death when given before 15 min of severe global ischemia achieved by hypotensive bilateral carotid clamping in rats. Ischemic blood flows less than 1% of the non-ischemic values were seen in the forebrain regions. Hypoperfusion occurred in all regions 60 min after the insult with flow values 21–58% of those in the non-ischemic group. Tirilazad had no effect on cerebral blood flow in the non-ischemic rats, nor in those decapitated during or after the insult. Five days postischemia neuronal damage had developed in all regions examined, but no significant differences were seen between the tirilazad-and the vehicle-treated rats.     

Effects of FO-1561 on postischemic cerebral functional and metabolic recovery in experimental cerebral ischemia

Effects of S-adenosyl-L-methionine sulfate tosylate (FO-1561) on postischemic cerebral functional and metabolic recovery in experimental cerebral ischemia were investigated. Severe bilateral forebrain ischemia in rats was induced by four-vessel occlusion with reducing the mean arterial pressure to 100-110 mmHg. After forebrain ischemia had been maintained for 30 minutes, recirculation was started by removal of the arterial clamps of bilateral common carotid arteries and by increasing systemic arterial pressure to the preischemic level. The EEG was continuously recorded from gold-coated screws inserted bilaterally in the parietal bones with the tips in extradural position, against a reference inserted prefrontal bone. Analysis of power spectrum of EEG activity was done by Berg Fourier Analyser. The brains were frozen in situ with liquid nitrogen before, during and after ischemia and then chiselled out during irrigation with liquid nitrogen. Concentrations of ATP in brain tissue were determined with high performance liquid chromatography. FO-1561, 100 mg/kg, was given intravenously, immediately after recirculation. After recirculation there was a tendency that EEG power spectrum in FO-1561-treated animals contained higher percentage of beta wave compared to that in control animals, while delta wave was lesser in FO-1561-treated animals. At 90 minutes following recirculation, ATP level in control animals was 2.17 +/- 0.05 mumol/g (mean +/- SE) and 2.42 +/- 0.03 mumol/g (mean +/- SE) in FO-1561-treated animals. Thus, recovery of ATP level was significantly better in FO-1561-treated animals than in control animals (p less than 0.01).     

Administration of selective endothelin receptor type A antagonist Ro 61-1790 does not improve outcome in focal cerebral ischemia in cat.

The authors examined the effect of selective endothelin (ET) receptor type A (ET(A)) antagonism on histological and functional recovery in cat at 24 hours after reversible middle cerebral artery occlusion (MCAO). A novel and specific ET(A) antagonist, Ro 61-1790 [5-methylpyridine-2-sulfonic acid-6-(2-hydroxyethoxy)-5-(2-methoxyphenoxy)-2-(2-1H-tetrazol-5-y l-pyridin-4-yl)-pyrimidin-4-ylamide sodium salt (1:2)] (Roche, Basel, Switzerland), was used at doses that produced steady-state plasma concentrations and abolished ET-induced pial arteriolar vasoconstriction. In a cranial window preparation, 8 nmol/L ET constricted pial arterioles by 33 +/- 18% (mean +/- SD), but this response was ablated by intravenous Ro 61-1790 treatment (10-mg/kg bolus, 4-mg/kg/h infusion). In additional animal cohorts, halothane-anesthetized cats were treated with 90 minutes of MCAO and 24 hours of reperfusion. Animals received Ro 61-1790 infusion beginning at the onset of reperfusion and continuing for 6 or 24 hours (n = 41). Control cats were treated with 0.9% saline by intravenous infusion throughout reperfusion. There was no difference in injury volume or neurologic evaluation score in saline-treated cats (n = 11; caudate 24 +/- 28%, cortical injury 7.5 +/- 5% of ipsilateral structure; score 52 +/- 8) versus the results in cats treated with Ro 61-1790 for either 24 hours (n = 6; caudate 22 +/- 23%, cortex 6 +/- 5%, injury volume of ipsilateral structure; score 55 +/- 3) or 6 hours (n = 11; caudate 33 +/- 30%, cortex 12 +/- 14%, injury volume of ipsilateral structure; score 50 +/- 10). Mortality was greatest in the 24-hour drug treatment group. These data suggest that blockade of ET(A) receptor activity is not beneficial to tissue or functional outcomes from experimental stroke in cat.     

Glatiramer Acetate administration does not reduce damage after cerebral ischemia in mice

Inflammation plays a key role in ischemic stroke pathophysiology: microglial/macrophage cells and type-1 helper cells (Th1) seem deleterious, while type-2 helper cells (Th2) and regulatory T cells (Treg) seem protective. CD4 Th0 differentiation is modulated by microglial cytokine secretion. Glatiramer Acetate (GA) is an immunomodulatory drug that has been approved for the treatment of human multiple sclerosis by means of a number of mechanisms: reduced microglial activation and pro-inflammatory cytokine production, Th0 differentiation shifting from Th2 to Th2 and Treg with anti-inflammatory cytokine production and increased neurogenesis.We induced permanent (pMCAo) or transient middle cerebral artery occlusion (tMCAo) and GA (2 mg) or vehicle was injected subcutaneously immediately after cerebral ischemia. Mice were sacrificed at D3 to measure neurological deficit, infarct volume, microglial cell density and qPCR of TNFα and IL-1β (pro-inflammatory microglial cytokines), IFNγ (Th2 cytokine), IL-4 (Th2 cytokine), TGFβ and IL-10 (Treg cytokines), and at D7 to evaluate neurological deficit, infarct volume and neurogenesis assessment.We showed that in GA-treated pMCAo mice, infarct volume, microglial cell density and cytokine secretion were not significantly modified at D3, while neurogenesis was enhanced at D7 without significant infarct volume reduction. In GA-treated tMCAo mice, microglial pro-inflammatory cytokines IL-1β and TNFα were significantly decreased without modification of microglial/macrophage cell density, cytokine secretion, neurological deficit or infarct volume at D3, or modification of neurological deficit, neurogenesis or infarct volume at D7.In conclusion, Glatiramer Acetate administered after cerebral ischemia does not reduce infarct volume or improve neurological deficit in mice despite a significant increase in neurogenesis in pMCAo and a microglial pro-inflammatory cytokine reduction in tMCAo.