ABSTRACT
The aim of the present study was to measure the therapeutic effects of bradykinin antagonists on lesion volume and brain swelling induced by cold injury in the parietal cortex of rat and mouse, respectively. Cold lesion was induced by application of a precooled (-78 degrees C) copper cylinder (3 mm diameter) to the intact dura of rat and mouse for 6 and 30 sec, respectively. At 24 h after the injury, the brains were removed and lesion volume was determined by the triphenyltetrazolium chloride method in rats. In the mouse, brain swelling was expressed as percentage increase in weight of the injured hemisphere which is compared to the contralateral side. After a subcutaneous priming dose of 18 microg/kg, a 1-h pretreatment and 24-h posttreatment using osmotic minipumps (300 ng/kg x min) was applied. Hoe140, a bradykinin receptor 2 antagonist, revealed a 19% reduction of lesion volume (p < 0.05) in the rat and a 14% diminution of brain swelling (p < 0.05) in the mouse. In contrast, the bradykinin receptor 1 antagonist, B 9858, had no effect on lesion volume compared to sham treated rats. When B 9858 was given in combination with Hoe140, a significant reduction in lesion volume was seen which was equivalent to and not different from that seen with Hoe140 alone in the rat. We conclude that brain injury after cold lesion is partially mediated by bradykinin and can be successfully treated with B2 antagonists.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Bradykinin Receptor Antagonists , Bradykinin/analogs & derivatives , Bradykinin/pharmacology , Brain Injuries/drug therapy , Neuroprotective Agents/pharmacology , Animals , Blood Pressure , Brain Edema/drug therapy , Brain Edema/pathology , Brain Injuries/pathology , Cold Temperature , Male , Mice , Rats , Rats, Inbred WKY , Receptor, Bradykinin B1 , Receptor, Bradykinin B2ABSTRACT
BACKGROUND AND PURPOSE: The present study was performed to determine the role of alpha4 (CD49d), a member of the integrin family of adhesion molecules, in ischemic brain pathology. METHODS: Male spontaneously hypertensive rats (SHR) or Sprague-Dawley rats underwent 60-minute middle cerebral artery occlusion (MCAO) followed by 23-hour reperfusion. Animals were injected intravenously with 2.5 mg/kg anti-rat alpha4 antibody (TA-2) or isotype control antibody (anti-human LFA-3 IgG(1), 1E6) 24 hours before MCAO. Infarct volume was quantified by staining of fresh tissue with tetrazolium chloride and myeloperoxidase activity measured in SHR tissue homogenates 24 hours after MCAO. In SHR, mean arterial blood pressure was recorded before and after MCAO in animals treated with TA-2 and 1E6. Fluorescence-activated cell sorting analysis was performed on peripheral blood leukocytes before and after MCAO. RESULTS: TA-2 treatment significantly reduced total infarct volume by 57.7% in normotensive rats (1E6, 84.2+/-11.5 mm(3), n=17; TA-2, 35.7+/-5.9 mm(3), n=16) and 35.5% in hypertensive rats (1E6, 146.6+/-15.5 mm(3), n=15; TA-2, 94.4+/-25.8 mm(3), n=11). In both strains, TA-2 treatment significantly reduced body weight loss and attenuated the hyperthermic response to MCAO. In SHR, treatment with TA-2 significantly reduced brain myeloperoxidase activity. Resting mean arterial blood pressure was unaffected by treatment. Leukocyte counts were elevated in TA-2-treated rats. Fluorescence-activated cell sorting analysis demonstrated the ability of TA-2 to bind to CD3+, CD4+, CD8+, and CD11b+ cells in both naive animals and after MCAO. CONCLUSIONS: These data demonstrate that inhibition of alpha4 integrin can protect the brain against ischemic brain injury and implicate endogenous alpha4 integrin in the pathogenesis of acute brain injury. The mechanism by which alpha4 integrin inhibition offers cerebroprotection is independent of blood pressure modulation and is likely due to inhibition of leukocyte function.
Subject(s)
Antigens, CD/metabolism , Cerebral Infarction/prevention & control , Ischemic Attack, Transient/metabolism , Animals , Antibodies/pharmacology , Antigens, CD/immunology , Antigens, CD/pharmacology , Blood Pressure/drug effects , Body Temperature/drug effects , Body Weight/drug effects , Brain/enzymology , Brain/pathology , Cell Adhesion Molecules/antagonists & inhibitors , Cell Adhesion Molecules/immunology , Cell Adhesion Molecules/metabolism , Cerebral Infarction/etiology , Cerebral Infarction/metabolism , Cerebral Infarction/pathology , Disease Models, Animal , Flow Cytometry , Infarction, Middle Cerebral Artery/complications , Integrin alpha4 , Ischemic Attack, Transient/etiology , Ischemic Attack, Transient/immunology , Ischemic Attack, Transient/pathology , Leukocyte Count , Leukocytes/cytology , Leukocytes/drug effects , Leukocytes/immunology , Male , Peroxidase/metabolism , Rats , Rats, Inbred SHR , Rats, Sprague-Dawley , Reproducibility of ResultsABSTRACT
BACKGROUND AND PURPOSE: Inflammation, a process that involves neutrophils, lymphocytes, and monocytes, contributes to cerebral ischemic injury. Blockade of neutrophil adhesion to endothelium improves outcome after experimental stroke. In this study we sought to assess the contribution of lymphocytes and monocytes to ischemic brain injury. METHODS: Male Lewis rats underwent 3 hours of middle cerebral artery occlusion followed by 45 hours of reperfusion. Two hours after the onset of ischemia, one group of animals received an intraperitoneal injection of antibodies to the alpha(4) integrin (n=16); another group was injected with an isotype control antibody (n=11). Neurological examination, body temperature, and body weight were assessed at different time points after stroke. Animals were killed 48 hours after the onset of ischemia for determination of infarct volume and leukocyte counts. RESULTS: There were no significant differences in body temperature or weight at any time. Neurological scores (deficits) were significantly less in animals treated with anti-alpha(4) antibodies at 24 (2.0+/-1.2 versus 3. 0+/-0.4; P:=0.006) and 48 (2.0+/-1.2 versus 3.0+/-0.8; P:=0.011) hours after ischemia. Peripheral blood leukocyte counts were significantly higher in anti-alpha(4)-treated animals (6.8+/-2.2 x 10(9) versus 2.9+/-1.9 x 10(9); P:=0.001) and revealed a lymphocyte/monocyte predominance (86.0+/-16.2% versus 71.0+/-15.6%; P:=0.008). Infarct volume was significantly less in animals treated with antibodies to alpha(4) (120.1+/-51.21 versus 173.7+/-42.29 mm(3); P:=0.012). CONCLUSIONS: These data support a role for lymphocytes and monocytes in cerebral ischemic injury and show that blockade of alpha(4), even when instituted after the onset of ischemia, can improve neurological outcome and decrease infarct volume.
Subject(s)
Antibodies/pharmacology , Antigens, CD/immunology , Cerebral Infarction/prevention & control , Ischemic Attack, Transient/immunology , Animals , Antigens, CD/metabolism , Body Temperature/drug effects , Body Weight/drug effects , Brain/drug effects , Brain/immunology , Brain/metabolism , Brain/pathology , Cell Adhesion Molecules/antagonists & inhibitors , Cell Adhesion Molecules/immunology , Cell Adhesion Molecules/metabolism , Cerebral Infarction/etiology , Cerebral Infarction/immunology , Cerebral Infarction/pathology , Infarction, Middle Cerebral Artery/complications , Integrin alpha4 , Ischemic Attack, Transient/etiology , Ischemic Attack, Transient/metabolism , Leukocyte Count , Lymphocytes/drug effects , Lymphocytes/immunology , Male , Monocytes/drug effects , Monocytes/immunology , Rats , Rats, Inbred Lew , Receptors, Lymphocyte Homing/antagonists & inhibitors , Receptors, Lymphocyte Homing/immunology , Receptors, Lymphocyte Homing/metabolism , Treatment OutcomeABSTRACT
The potential use of anti-alpha4 antibody presents an attractive approach to the treatment of stroke for a number of reasons: firstly, access to the site of action; alpha(4) is expressed on circulating leukocytes, permitting efficacy with systemic administration and negating the requirement for passage across the blood-brain barrier, an important consideration for neuroprotective and neurotrophic agents and other antiinflammatory approaches such as chemokine inhibitors. Pharmacokinetic studies in the rat have demonstrated a maintained pharmacological action of alpha(4) antibody for up to 72 hours after a single intravenous dose using leukocytosis as a surrogate marker for activity. Finally, the ubiquitous pattern of alpha(4) expression on different leukocyte subsets makes it an attractive target to combat the early and late inflammatory responses to brain injury.
ABSTRACT
The physical parameters governing adsorption of DNA by various positively charged depth filters and membranes have been assessed. Buffers that reduced or neutralised the depth filter or membrane charge, and those that impeded hydrophobic interactions were shown to affect their operational capacity, demonstrating that DNA was adsorbed by a combination of electrostatic and hydrophobic interactions. The adsorption profile of DNA by a Sartobind Q anion exchange membrane showed immediate breakthrough, irrespective of challenge DNA concentration or flow rate, and in this case adsorption was by electrostatic interactions only. The production-scale removal of DNA from harvest broths containing therapeutic protein by partitioning of cells and debris from protein in sequential centrifugation and filtration steps, and the concentration of DNA in process supernatant were assessed. Centrifugation reduced the quantity of DNA in the process material from 79.8 micrograms ml-1 to 9.3 micrograms ml-1 whereas the concentration of DNA in the supernatant of pre- and post-filtration samples had only marginally reduced DNA content: from 6.3 to 6.0 micrograms ml-1 respectively. DNA was concentrated to 27.3 micrograms ml-1 along with monoclonal antibody in the ultrafiltration step. Similar effects were observed in the harvest step for a second antibody.
Subject(s)
Antibodies, Monoclonal/biosynthesis , DNA/immunology , Filtration/instrumentation , Membranes, Artificial , Adsorption , Animals , Cattle , DNA/isolation & purificationABSTRACT
BACKGROUND AND PURPOSE: Recent studies demonstrated a significant neuroprotective action of the selective peptide-based bradykinin B2 receptor antagonist CP-0597 after permanent middle cerebral artery (MCA) occlusion (MCAO) in the rat. We therefore evaluated the efficacy of this compound after reversible MCAO in the rat. METHODS: Male Wistar rats underwent reversible MCAO by insertion of a nylon monofilament to the origin of the MCA. After 1 hour the filament was retracted and the ischemic tissue reperfused. Immediately after MCAO, primed miniosmotic pumps containing either vehicle or CP-0597 (300 ng/kg per minute) were implanted into the subcutaneous space (n = 14 per group). Twenty-four hours after surgery, animals were killed and brains fixed, and 4-micron sections were taken from five sequential tissue blocks labeled A through E and stained with hematoxylin and eosin. Clinical evaluation of rats was performed by neurological scoring and change in body weight. RESULTS: Treatment with CP-0597 significantly reduced percent increase in hemisphere size of the ischemic hemisphere in all brain sections (C section: vehicle, 40.6 +/- 4.3% versus CP-0597, 20.8 +/- 5.3%; P < 0.001), total infarct volume (vehicle, 206.5 +/- 7.7 mm3 versus CP-0597, 94.0 +/- 19.2 mm3; P < .001), cortical infarct volume (vehicle, 145.5 +/- 4.5 mm3 versus CP-0597, 64.0 +/- 15.1 mm3; P < .001), subcortical infarct volume (vehicle, 55.8 +/- 4.1 mm3 versus CP-0597, 27.5 +/- 4.5 mm3; P < .001), and the number of necrotic neurons (vehicle 42.9 +/- 3.8 versus CP-0597, 23.6 +/- 4.7 per field; P < .01). Neurological score (vehicle, 2.78 +/- 0.36 versus CP-0597, 6.29 +/- 0.87 P < .01) and change in body weight (vehicle, -28.7 +/- 2.0 g versus CP-0597, -18.2 +/- 2.8 g; P < .01) were also significantly improved. CONCLUSIONS: The present data demonstrate the significant overall efficacy profile of CP-0597 in a rat model of reversible MCAO and provide strong rationale for the use of such bradykinin B2 receptor antagonist in the treatment of stroke.
Subject(s)
Arterial Occlusive Diseases/drug therapy , Bradykinin Receptor Antagonists , Brain Ischemia/drug therapy , Brain/blood supply , Cerebral Arteries , Oligopeptides/pharmacology , Reperfusion Injury/drug therapy , Animals , Arterial Occlusive Diseases/metabolism , Behavior, Animal/drug effects , Body Temperature , Body Weight , Brain Chemistry/physiology , Brain Edema/drug therapy , Brain Edema/metabolism , Brain Ischemia/metabolism , Cerebrovascular Disorders/drug therapy , Cerebrovascular Disorders/metabolism , Disease Models, Animal , Male , Rats , Rats, Wistar , Receptor, Bradykinin B2 , Reperfusion Injury/metabolismABSTRACT
Interleukin-1 beta is a proinflammatory cytokine released by activated immune cells. In addition to orchestrating immune responses to infection, interleukin-1 beta is a key mediator of immune-to-brain communication. Interleukin-1 beta and endotoxin (which releases IL1 beta from immune cells) cause centrally mediated illness responses such as fever, aphagia, etc. These effects are blocked by intraperitoneal IL1 receptor antagonist (IL1ra), suggesting critical involvement of peripheral IL1 receptors. Centrally mediated illness responses are also blocked by vagotomy, suggesting that IL1 beta directly or indirectly activates vagal afferents. To test for IL1 beta binding whole vagus (abdominal, laryngeal, and thoracic) and sections of hepatic vagus and liver hilus were incubated with biotinylated IL1ra and processed for avidin-biotin complex (ABC) or avidin-FITC histochemistry. Glomus cells of vagal paraganglia were labeled in all regions of the vagus. Biotinylated IL1ra also labeled smooth muscle and endothelial cells of blood vessels and lymphoid tissues. No label was present in omission or competition controls. These data suggest that centrally mediated illness responses result from IL1 activation of vagal paraganglia.
Subject(s)
Interleukin-1/pharmacology , Receptors, Interleukin-1/antagonists & inhibitors , Vagus Nerve/drug effects , Animals , Dose-Response Relationship, Drug , Histocytochemistry , Male , Rats , Rats, Sprague-DawleyABSTRACT
Recombinant human ciliary neurotrophic factor (rh-CNTF) was reported to attenuate skeletal muscle wasting in rats after unilateral transection of the sciatic nerve (M. E. Helgren, S. P. Squinto, H. L. Davis, D. J. Parry, T. G. Bolton, C. S. Heck, Y. Zhu, G. D. Yancopoulos, R. M. Lindsay, and P. S. DiStefano. Cell 76: 493-504, 1994). Under the experimental conditions reported herein, the absolute masses of the denervated gastrocnemius and soleus muscles were not increased in mature or immature rats of either sex by treatment with rhCNTF. At the highest doses of rhCNTF (1 and 0.1 mg/kg), increases in the ratio of the masses of the denervated to the contralateral innervated gastrocnemius and soleus muscles could be attributed entirely to a muscle-wasting effect on the contralateral innervated muscle rather than any muscle-sparing effect on the denervated muscle. The muscle-wasting effects of rhCNTF were associated with reductions in body weight gain and reduced food intake. Pair-fed rats lost less body weight and skeletal muscle mass than rhCNTF-injected freely fed rats but experienced significantly greater loss of visceral mass. Male rats displayed greater loss of body weight and skeletal muscle mass than female rats. Recombinant inhibitors of the cachectic cytokines, tumor necrosis factor and interleukin-1, did not significantly alter the wasting effects of rhCNTF. These findings demonstrate that, in contrast to its well-characterized trophic effects on cells of the nervous system, rhCNTF causes atrophy of skeletal muscle by mechanisms involving both anorexia and cachexia based on the results of pair-feeding experiments.
Subject(s)
Cachexia/physiopathology , Muscle, Skeletal/innervation , Muscle, Skeletal/physiopathology , Nerve Tissue Proteins/pharmacology , Animals , Body Weight/drug effects , Ciliary Neurotrophic Factor , Cytokines/antagonists & inhibitors , Denervation , Female , Humans , Male , Muscular Atrophy/etiology , Muscular Atrophy/prevention & control , Nervous System/physiopathology , Organ Size/drug effects , Rats , Rats, Sprague-Dawley , Recombinant ProteinsABSTRACT
We assessed the efficacy of recombinant human interleukin-1 receptor antagonist (rhIL-1ra) on brain injury and edema formation after permanent middle cerebral artery occlusion (MCAo) in the rat. Previous studies showed that low amounts of rhIL-1ra injected directly into the brain significantly decreased infarct size after MCAo or excitotoxic injury in rats. Peripheral administration of rhIL-1ra (100 mg/kg sc at 0, 4, 8, 12, and 18 h after MCAo) significantly inhibited infarct size, by 46% (P < 0.05), measured at 24h. This was greater than the effect of MK801 administered immediately after MCAo (4 mg/kg ip, 0 h) which did not significantly reduce infarct size. rhIL-1ra (100 mg/kg also significantly inhibited cerebral edema formation by 49% (p< 0.05 measured 24 h after MCAo, but did not reduce edema formation measured 2 h after MCAo, but did not reduce edema formation measured 2 h after MCAo. Inhibition of infarction by rhIL-1ra was dependent on dose and time of administration. Together the results demonstrate that peripherally administered rhIL-1ra at high doses is able to mimic the efficacy of low dose of rhIL-1ra administered directly into the brain in a rodent model of stroke and that protection observed with rhIL-1ra was better than that offered by MK801 in this model.
Subject(s)
Brain Damage, Chronic/prevention & control , Brain Ischemia/complications , Receptors, Interleukin-1/antagonists & inhibitors , Animals , Blood Glucose/analysis , Brain Edema/prevention & control , Cerebral Infarction/pathology , Dizocilpine Maleate/pharmacology , Humans , Injections, Intravenous , Injections, Subcutaneous , Male , Rats , Rats, Sprague-Dawley , Recombinant ProteinsABSTRACT
Marked increases in the brain expression of interleukin (IL)-1 have been reported in rats after permanent occlusion of a large cerebral artery. Interactions between endothelial cells and leukocytes have been implicated in the pathogenesis of several types of ischemic injury to the myocardium and other organs. In this study we asked whether inhibiting the effects of IL-1 would affect the outcome of an experimental brain infarct. Adult male Wistar rats (n = 13) with permanent occlusion of the middle cerebral artery were given IL-1 receptor antagonist. A second group (n = 13) with the same type of brain injury was given a placebo. A third group, subjected to a sham operation, was given either IL-1 receptor antagonist (n = 2) or a placebo (n = 2). Experiments were terminated after either 24 hours or 7 days. Compared with the control group, animals treated with IL-1 receptor antagonist improved their neurological score (P < 0.05), experienced less pronounced changes in body weight (P < 0.05), and had fewer necrotic neurons (P < 0.001) and fewer leukocytes in the ischemic hemisphere (P < 0.001) as well as a smaller area of pallor (P < 0.05) in the ischemis hemisphere. The results suggest that inhibiting the proinflammatory effects of IL-1 with a receptor antagonist is an effective way of influencing the leukocyte responses elicited by an arterial occlusion. Such leukocyte inhibition seemingly attenuates the number of necrotic neurons resulting from the occlusion of a large brain artery.
Subject(s)
Arterial Occlusive Diseases/pathology , Cerebral Arteries , Neurons/pathology , Sialoglycoproteins/therapeutic use , Animals , Arterial Occlusive Diseases/drug therapy , Arterial Occlusive Diseases/physiopathology , Body Weight/drug effects , Brain Ischemia/drug therapy , Brain Ischemia/pathology , Brain Ischemia/physiopathology , Cerebral Cortex/ultrastructure , Glial Fibrillary Acidic Protein/immunology , Interleukin 1 Receptor Antagonist Protein , Interleukin-1/antagonists & inhibitors , Male , Necrosis , Neurons/drug effects , Neurons/ultrastructure , Neutrophils/pathology , Neutrophils/ultrastructure , Rats , Rats, WistarABSTRACT
Activation of immune cells by pathogens induces the release of a variety of proinflammatory cytokines, including IL-1 beta and TNF-alpha. Previous studies using IL-1 beta have demonstrated that this cytokine can alter brain function, resulting in a variety of 'illness responses' including increased sleep, decreased food intake, fever, etc. We have recently demonstrated that i.p. IL-1 beta also produces hyperalgesia and that this hyperalgesia (as well as most illness responses) is mediated via activation of subdiaphragmatic vagal afferents. The present series of studies were designed to provide an initial examination of the generality of proinflammatory cytokine-induced hyperalgesia by examining the effects of i.p. TNF-alpha on pain responsivity. These studies demonstrate that: (a) i.p. TNF-alpha produces dose-dependent hyperalgesia as measured by the tailflick test, (b) this hyperalgesia is mediated via the induced release of IL-1 beta, (c) hyperalgesia is mediated via activation of subdiaphragmatic vagal afferents, and (d) the effects of subdiaphragmatic vagotomy cannot be explained by a generalized depression of neural excitability.
Subject(s)
Hyperalgesia/chemically induced , Tumor Necrosis Factor-alpha/toxicity , Animals , Dose-Response Relationship, Drug , Formaldehyde , Humans , Hyperalgesia/physiopathology , Injections, Intraperitoneal , Interleukin-1/administration & dosage , Interleukin-1/pharmacology , Male , Pain Threshold/drug effects , Rats , Rats, Sprague-Dawley , Receptors, Interleukin-1/antagonists & inhibitors , Recombinant Proteins/toxicity , Vagotomy , Vagus Nerve/physiopathologyABSTRACT
Interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) are cytokines released by activated immune cells. IL-1 beta and TNF-alpha elicit various illness symptoms including avoidance of novel tastes with which they have been paired (conditioned taste aversion). Previous hypotheses to account for these actions have focused on blood-borne IL-1 beta and TNF-alpha exerting their effects directly at the brain. However, recent evidence suggests that these cytokines may activate subdiaphragmatic vagal afferents. The present experiments demonstrate that subdiaphragmatic vagal transection both attenuates acquisition and facilitates extinction of conditioned taste aversions induced by i.p. administration of either IL-1 beta or TNF-alpha.
Subject(s)
Brain/immunology , Taste , Vagus Nerve/physiology , Animals , Cytokines/pharmacology , Interleukin-1/pharmacology , Male , Rats , Rats, Sprague-Dawley , Saccharin , Tumor Necrosis Factor-alpha/pharmacology , Vagotomy , Vagus Nerve/immunologyABSTRACT
Processes occurring within the immune system can alter neural function. Cytokines released by cells of the immune system during illness are key messengers in immune-to-brain communication. Interleukin-1 beta (IL-1 beta) is particularly important in this regard and is known to stimulate a myriad of illness-related outcomes such as fever, sickness behavior, aphagia, adipsia, hypothalamic-pituitary-adrenal activation, and changes in pain reactivity. Thus peripherally released IL-1 beta has potent neural effects and is a critical mediator of the impact of immune processes on brain. There is, however, uncertainty concerning the communication pathways involved. We provide evidence that a primary route of peripheral cytokine signalling is through stimulation of peripheral vagal afferents rather than or in addition to direct cytokine access to brain. Subdiaphragmatic, but not hepatic vagotomy, blocked rhIL-1 beta-induced hypothalamic norepinephrine depletion and attenuated rhIL-1 beta-induced increases in serum corticosterone. These data suggest that rhIL-1 beta activates the hypothalamic-pituitary-adrenal axis via stimulation of peripheral vagal afferents and further support the hypothesis that peripheral cytokine signalling to the CNS is mediated primarily by stimulation of peripheral afferents.
Subject(s)
Corticosterone/blood , Hypothalamus/metabolism , Interleukin-1/pharmacology , Norepinephrine/metabolism , Vagus Nerve/physiology , Animals , Diaphragm/innervation , Dose-Response Relationship, Drug , Hypothalamus/drug effects , Liver/innervation , Male , Rats , Rats, Sprague-Dawley , Recombinant Proteins/pharmacology , VagotomyABSTRACT
Interleukin-1 beta (IL-1 beta), a cytokine released by activated immune cells, elicits various illness symptoms including hyperthermia. Previous hypotheses to account for these actions have focused on blood-borne IL-1 beta exerting its effects directly at the level of the brain. However, recent behavioral and physiological evidence suggest that IL-1 beta can activate the subdiaphragmatic vagus. The present experiments demonstrate that subdiaphragmatic vagal transection disrupts the hyperthermia-inducing effects of recombinant human IL-1 beta and stress. These data provide evidence for a novel route of immune-brain communication, as well as a novel route whereby stress can influence physiological processes.
Subject(s)
Brain/immunology , Fever , Interleukin-1/pharmacology , Animals , Male , Rats , Rats, Sprague-Dawley , Stress, Physiological , Time Factors , VagotomyABSTRACT
This study investigated the involvement of corticotrophin-releasing factor (CRF) in acute neuronal damage induced by focal cerebral ischaemia or pharmacological activation of NMDA receptors in the rat brain. Intracerebroventricular injection of a CRF receptor antagonist (alpha-helical CRF9-41), markedly inhibited ischaemic (61%) and excitotoxic (41%) brain damage. Peripheral injection of a glucocorticoid antagonist (RU38486) did not affect ischaemic damage. Ischaemic and excitotoxic damage caused increased hypothalamic concentrations of CRF. These data indicate that CRF mediates ischaemic and excitotoxic neuronal damage in the rat, but that this effect is not dependent on glucocorticoids.
Subject(s)
Brain/metabolism , Corticotropin-Releasing Hormone/antagonists & inhibitors , Corticotropin-Releasing Hormone/pharmacology , Ischemic Attack, Transient/pathology , Neurons/drug effects , Peptide Fragments/pharmacology , Receptors, N-Methyl-D-Aspartate/metabolism , Animals , Body Temperature/drug effects , Brain/drug effects , Corticotropin-Releasing Hormone/administration & dosage , Corticotropin-Releasing Hormone/metabolism , Hypothalamus/drug effects , Hypothalamus/metabolism , Hypothalamus/pathology , Injections, Intraventricular , Male , Mifepristone/administration & dosage , Mifepristone/pharmacology , Peptide Fragments/administration & dosage , Rats , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate/antagonists & inhibitors , Receptors, N-Methyl-D-Aspartate/drug effectsABSTRACT
Arachidonic acid [20:4(N-6)] has been implicated in neurological damage induced by cerebral ischaemia. Membrane arachidonate concentrations can be reduced by changes in dietary fat intake. Therefore, in the present study, we have investigated the effects of N-3 fatty acid supplementation on neuronal damage induced by permanent focal cerebral ischaemia or pharmacological activation of striatal NMDA receptors. Weanling rats were fed either a control diet or an N-3 supplemented diet (1.75% by weight as N-3 fatty acids) for 6 weeks. N-3-supplemented animals reduced ischaemic damage following middle cerebral artery occlusion (36%, p < 0.05), and excitotoxic damage induced by infusion of the selective NMDA agonist (1-aminocyclobutane-cis-1,3-dicarboxylic acid, 43%, p < 0.001) compared to the control-fed group. These data are consistent with the proposed role of arachidonic acid in ischaemic and excitotoxic brain damage, and suggest that modest dietary supplementation with N-3 fatty acids may offer benefit to populations at high risk of stroke.
Subject(s)
Amino Acids, Cyclic , Amino Acids/toxicity , Brain/drug effects , Dietary Fats/pharmacology , Fatty Acids, Omega-3/pharmacology , Ischemic Attack, Transient/prevention & control , Neurons/drug effects , Neurons/pathology , Neurotoxins/toxicity , Animals , Brain/pathology , Corpus Striatum/drug effects , Food, Fortified , Infusions, Parenteral , Ischemic Attack, Transient/pathology , Male , Rats , Rats, Sprague-DawleyABSTRACT
The cytokine interleukin-1 (IL-1) is synthesised within the brain and acts as a mediator of host defence responses to disease and injury. Several of these central actions of IL-1 are inhibited by an endogenous calcium and phospholipid binding protein, lipocortin-1. Synthesis of IL-1 and lipocortin-1 in the brain is markedly increased by neuronal damage, and inhibition of the actions of endogenous IL-1 by central injection of IL-1 receptor antagonist in the rat significantly inhibits ischaemic and excitotoxic brain damage. Lipocortin-1 appears to act as an endogenous neuroprotective agent that markedly attenuates ischaemic and excitotoxic damage. In contrast, inhibition of the actions of lipocortin-1 by injection of neutralising antiserum exacerbates both forms of neurodegeneration. The mechanisms underlying these effects of IL-1 and lipocortin-1 are largely unknown, but are probably independent of changes in body temperature. Actions of these molecules on corticotrophin releasing factor, arachidonic acid, excitatory amino acids, and nitric oxide, and the possible involvement of these factors in brain damage are discussed.
Subject(s)
Annexin A1/physiology , Brain Ischemia/physiopathology , Interleukin-1/physiology , Animals , Brain Ischemia/pathology , HumansABSTRACT
Cytokines, (particularly interleukins and growth factors) are synthesised in the brain, and induced by brain damage. Interleukin-I appears to directly mediate ischaemic and excitotoxic brain damage, whereas growth factors (e.g., bFGF, NGF), and the phospholipid binding protein lipocortin-1 exhibit neuroprotective actions. Central administration of recombinant interleukin-1 receptor antagonist markedly attenuates damage induced by focal cerebral ischaemia, or pharmacological activation of NMDA receptors in the rat brain. The mechanisms of action of these cytokines on neurodegeneration are unknown, but indirect evidence has implicated corticotropin releasing factor, arachidonic acid, and nitric oxide. In vitro effects of interleukin-1, growth factors, and lipocortin-1 have been reported on intracellular calcium homeostasis, which is critically important in neurodegeneration. Pharmacological modulation of the expression and/or actions of cytokines in the brain may be of considerable therapeutic benefit in the treatment of acute neurodegeneration.
Subject(s)
Central Nervous System/physiology , Cytokines/physiology , Nerve Degeneration/physiology , Animals , HumansABSTRACT
In the present studies, the mechanisms underlying the inhibitory actions of lipocortin-1 on the pyrogenic and thermogenic properties of cytokines were investigated. Central (icv) injection of corticotropin-releasing factor (CRF, 4.7 micrograms) or the recombinant cytokines interleukin (IL)-1 alpha (50 ng), IL-1 beta (5 ng), IL-6 (20 ng), IL-8 (20 ng), or tumor necrosis factor-alpha (TNF-alpha, 1 microgram) in conscious rats produced significant increases in resting oxygen consumption (VO2, 13-26%) and colonic temperature (0.7-1.6 degrees C) within 2 h postinjection. Administration (icv) of a recombinant fragment (NH2-terminus, 1-188 amino acids) of human lipocortin-1 (1.2 micrograms) produced small increases in VO2 (< 5%) and body temperature (< 0.3 degrees C). Pretreatment (-5 min) with lipocortin-1 significantly attenuated the thermogenic and pyrogenic effects of centrally injected IL-1 beta (80% inhibition), IL-6 (60%), IL-8 (80%), or CRF (60%). However, pretreatment with lipocortin-1 failed to modify the actions of IL-1 alpha or TNF-alpha. We have previously demonstrated that the pyrogenic and thermogenic effects of IL-1 beta, IL-6, and IL-8 are dependent on the central actions of CRF, whereas IL-1 alpha and TNF-alpha act independently of CRF. Fever and thermogenesis induced by all of these cytokines (with the exception of IL-8) can also be prevented by administration of a cyclooxygenase inhibitor. The data presented here suggest that the potent antipyretic effects of lipocortin-1 may result from inhibition of the release or actions of CRF rather than modulation of eicosanoid synthesis.