phosphocreatine and Hyperglycemia

Hyperglycemia due to relative hypoinsulinism is common in extremely preterm infants and is associated with hippocampus-mediated long-term cognitive impairment. In neonatal rats, hypoinsulinemic hyperglycemia leads to oxidative stress, altered neurochemistry, microgliosis, and abnormal synaptogenesis in the hippocampus. Intranasal insulin (INS) bypasses the blood-brain barrier, targets the brain, and improves synaptogenesis in rodent models, and memory in adult humans with Alzheimer's disease or type 2 diabetes, without altering the blood levels of insulin or glucose. To test whether INS improves hippocampal development in neonatal hyperglycemia, rat pups were subjected to hypoinsulinemic hyperglycemia by injecting streptozotocin (STZ) at a dose of 80 mg/kg i.p. on postnatal day (P) 2 and randomized to INS, 0.3U twice daily from P3-P6 (STZ + INS group), or no treatment (STZ group). The acute effects on hippocampal neurochemical profile and transcript mRNA expression of insulin receptor (Insr), glucose transporters (Glut1, Glut4, and Glut8), and poly(ADP-ribose) polymerase-1 (Parp1, a marker of oxidative stress) were determined on P7 using in vivo 1H MR spectroscopy (MRS) and qPCR. The long-term effects on the neurochemical profile, microgliosis, and synaptogenesis were determined at adulthood using 1H MRS and histochemical analysis. Relative to the control (CONT) group, mean blood glucose concentration was higher from P3 to P6 in the STZ and STZ + INS groups. On P7, MRS showed 10% higher taurine concentration in both STZ groups. qPCR showed 3-folds higher Insr and 5-folds higher Glut8 expression in the two STZ groups. Parp1 expression was 18% higher in the STZ group and normal in the STZ + INS group. At adulthood, blood glucose concentration in the fed state was higher in the STZ and STZ + INS groups. MRS showed 59% higher brain glucose concentration and histochemistry showed microgliosis in the hippocampal subareas in the STZ group. Brain glucose was normal in the STZ + INS group. Compared with the STZ group, phosphocreatine and phosphocreatine/creatine ratio were higher, and microglia in the hippocampal subareas fewer in the STZ + INS group (p < 0.05 for all). Neonatal hyperglycemia was associated with abnormal glucose metabolism and microgliosis in the adult hippocampus. INS administration during hyperglycemia attenuated these adverse effects and improved energy metabolism in the hippocampus.

Topics: Adult; Animals; Blood Glucose; Diabetes Mellitus, Type 2; Glucose; Hippocampus; Humans; Hyperglycemia; Infant, Newborn; Infant, Premature; Insulin; Phosphocreatine; Rats; Streptozocin

Diabetic ketoacidosis (DKA) may cause brain injuries in children. The mechanisms responsible are difficult to elucidate because DKA involves multiple metabolic derangements. We aimed to determine the independent effects of hyperglycemia and ketosis on cerebral metabolism, blood flow, and water distribution. We used magnetic resonance spectroscopy to measure ratios of cerebral metabolites (ATP to inorganic phosphate [Pi], phosphocreatine [PCr] to Pi, N-acetyl aspartate [NAA] to creatine [Cr], and lactate to Cr) and diffusion-weighted imaging and perfusion-weighted imaging to assess cerebral water distribution (apparent diffusion coefficient [ADC] values) and cerebral blood flow (CBF) in three groups of juvenile rats (hyperglycemic, ketotic, and normal control). ATP-to-Pi ratio was reduced in both hyperglycemic and ketotic rats in comparison with controls. PCr-to-Pi ratio was reduced in the ketotic group, and there was a trend toward reduction in the hyperglycemic group. No significant differences were observed in NAA-to-Cr or lactate-to-Cr ratio. Cortical ADC was reduced in both groups (indicating brain cell swelling). Cortical CBF was also reduced in both groups. We conclude that both hyperglycemia and ketosis independently cause reductions in cerebral high-energy phosphates, CBF, and cortical ADC values. These effects may play a role in the pathophysiology of DKA-related brain injury.

Topics: Adenosine Triphosphate; Animals; Aspartic Acid; Brain Edema; Cerebrum; Diabetes Mellitus, Experimental; Diabetic Ketoacidosis; Diet, High-Fat; Hyperglycemia; Lactic Acid; Magnetic Resonance Spectroscopy; Phosphates; Phosphocreatine; Rats; Water

As recently demonstrated by our group (da-Silva, W. S., Gómez-Puyou, A., Gómez-Puyou, M. T., Moreno-Sanchez, R., De Felice, F. G., de Meis, L., Oliveira, M. F., and Galina, A. (2004) J. Biol. Chem. 279, 39846-39855) mitochondrial hexokinase activity (mt-HK) plays a preventive antioxidant role because of steady-state ADP re-cycling through the inner mitochondrial membrane in rat brain. In the present work we show that ADP re-cycling accomplished by the mitochondrial creatine kinase (mt-CK) regulates reactive oxygen species (ROS) generation, particularly in high glucose concentrations. Activation of mt-CK by creatine (Cr) and ATP or ADP, induced a state 3-like respiration in isolated brain mitochondria and prevention of H(2)O(2) production obeyed the steady-state kinetics of the enzyme to phosphorylate Cr. The extension of the preventive antioxidant role of mt-CK depended on the phosphocreatine (PCr)/Cr ratio. Rat liver mitochondria, which lack mt-CK activity, only reduced state 4-induced H(2)O(2) generation when 1 order of magnitude more exogenous CK activity was added to the medium. Simulation of hyperglycemic conditions, by the inclusion of glucose 6-phosphate in mitochondria performing 2-deoxyglucose phosphorylation via mt-HK, induced H(2)O(2) production in a Cr-sensitive manner. Simulation of hyperglycemia in embryonic rat brain cortical neurons increased both DeltaPsi(m) and ROS production and both parameters were decreased by the previous inclusion of Cr. Taken together, the results presented here indicate that mitochondrial kinase activity performed a key role as a preventive antioxidant against oxidative stress, reducing mitochondrial ROS generation through an ADP-recycling mechanism.

Topics: Adenosine Diphosphate; Animals; Antioxidants; Brain; Cells, Cultured; Creatine; Creatine Kinase, Mitochondrial Form; Hydrogen Peroxide; Hyperglycemia; In Vitro Techniques; Male; Membrane Potential, Mitochondrial; Mitochondria; Models, Biological; Oxygen Consumption; Phosphocreatine; Rats; Rats, Wistar; Reactive Oxygen Species; Substrate Cycling

Iron-catalyzed radical generation is a potentially significant mechanism by which extensive tissue acidosis exacerbates brain injury during ischemia/reperfusion. We hypothesized that levels of low-molecular-weight (LMW) iron increase during in vivo global cerebral ischemia in a pH-dependent manner, potentially catalyzing oxidant injury. The present study quantified regional differences in LMW iron during global cerebral incomplete ischemia and determined whether augmenting the fall in ischemic tissue pH with hyperglycemia also amplifies free iron availability.. Dogs anesthetized with pentobarbital-fentanyl were treated with 30 minutes of global incomplete cerebral ischemia produced by intracranial pressure elevation. Cerebral energy metabolites (ATP, phosphocreatine) and intracellular pH (pHi) were measured by 31P magnetic resonance spectroscopy. Preischemic plasma glucose level was manipulated to titrate end-ischemic pHi. After ischemia, brains were perfused with cold phosphate-buffered saline solution; then 16 different brain areas were sampled, filtered to separate the LMW fraction (<30000 D), and assayed by rapid colorimetric assay for tissue iron. Total iron, LMW iron, and protein in each sample were measured in sham-operated (no ischemia, n=8), normoglycemic ischemia (ISCH [glucose 7+/-4 mmol/L], n=7), and hyperglycemic (GLU-ISCH [glucose 31+/-3 mmol/L], n=9) groups.. High-energy phosphates fell to near zero values in both ISCH and GLU-ISCH groups by 30 minutes but remained unchanged in the sham-operated group. As expected, pHi decreased during ischemia but to a greater extent in GLU-ISCH (6.20+/-0.05 in ISCH, 6.08+/-0.04 in GLU-ISCH, P<.05). Iron could be detected in all areas of the brain in sham-operated animals, with the highest amounts obtained from subcortical areas such as the hippocampus, pons, midbrain, and medulla. Total iron was higher in ISCH relative to sham-operated animals and higher in cortex and pons relative to GLU-ISCH. Regional LMW (as a percentage of total iron; LMW/total iron) was elevated in numerous brain areas in ISCH, including cortical gray matter, cerebellum, hippocampus, caudate, and midbrain. LMW/total iron was higher in GLU-ISCH versus ISCH in cortical gray matter only. In other brain areas, ischemic LMW/total iron was equivalent in glucose-treated or normoglycemic animals (white matter, thalamus, pons, medulla) or lower in the glucose-treated group (cerebellum, hippocampus, caudate, midbrain).. These data demonstrate that levels of total and LMW iron increase with global cerebral ischemia in the majority of cortical and subcortical regions of normoglycemic brain. However, exacerbation of ischemic acidosis via glucose administration does not increase tissue iron and produces a greater increase in the LMW fraction in cortical gray matter only. In other brain regions, total and LMW iron availability is similar to that of nonischemic animals.

Topics: Acidosis; Adenosine Triphosphate; Animals; Blood Glucose; Blood Pressure; Body Temperature; Brain; Carbon Dioxide; Dogs; Hydrogen-Ion Concentration; Hyperglycemia; Intracranial Pressure; Iron; Ischemic Attack, Transient; Male; Organ Specificity; Oxygen; Partial Pressure; Phosphocreatine; Reference Values; Time Factors

The effects of hyperglycemia on brain pyruvate dehydrogenase (PDH) and metabolites (ATP, PCr, and lactate) were investigated at 20 min ischemia, 0, 20, and 60 min, and 4 h reperfusion. During reperfusion, PDH activities were suppressed corresponding to the poor recovery of ATP and PCr concentrations and the increase in lactate concentration in the hyperglycemic group, suggesting that preischemic hyperglycemia may impair metabolism by suppressing PDH activity.

Topics: Adenosine Triphosphate; Animals; Cerebral Cortex; Energy Metabolism; Gerbillinae; Hyperglycemia; Ischemic Attack, Transient; Lactic Acid; Male; Phosphocreatine; Pyruvate Dehydrogenase Complex; Reperfusion

The purpose of this study was to test the hypothesis that hyperglycemia ameliorates changes in brain cell membrane function and preserves cerebral high energy phosphates during hypoxia-ischemia in newborn piglets. A total of 42 ventilated piglets were divided into 4 groups, normoglycemic/normoxic(group 1, n=9), hyperglycemic/normoxic(group 2, n=8), normoglycemic/hypoxic-ischemic(group 3, n=13) and hyperglycemic/hypoxic-ischemic(group 4, n=12) group. Cerebral hypoxia-ischemia was induced by occlusion of bilateral common carotid arteries and simultaneous breathing with 8% oxygen for 30 min. Hyperglycemia (blood glucose 350-400 mg/dl) was maintained for 90 min before and throughout hypoxia-ischemia using modified glucose clamp technique. Changes in cytochrome aa3 were continuously monitored using near infrared spectroscopy. Blood and CSF glucose and lactate were monitored. Na+, K+-ATPase activity, lipid peroxidation products (conjugated dienes), tissue high energy phosphates (ATP and phosphocreatine) levels and brain glucose and lactate levels were determined biochemically in the cerebral cortex. During hypoxia-ischemia, glucose levels in blood and CSF were significantly elevated in hyperglycemic/hypoxic-ischemic group compared with normoglycemic/hypoxic-ischemic group, but lactate levels in blood and CSF were not different between two groups. At the end of hypoxia-ischemia of group 3 and 4, triangle up Cyt aa3, Na+, K+-ATPase activity, ATP and phosphocreatine values in brain were significantly decreased compared with normoxic groups 1 and 2, but were not different between groups 3 and 4. Levels of conjugated dienes and brain lactate were significantly increased in groups 3 and 4 compared with groups 1 and 2, and were significantly elevated in group 4 than in group 3 (0.30+/-0.11 vs. 0.09+/-0.02 micromol g-1 protein, 26.4+/-7.6 vs. 13.1+/-2.6 mmol kg-1, p<0.05). These findings suggest that hyperglycemia does not reduce the changes in brain cell membrane function and does not preserve cerebral high energy phosphates during hypoxia-ischemia in newborn piglets. We speculate that hyperglycemia may be harmful during hypoxia-ischemia due to increased levels of lipid peroxidation in newborn piglet.

Topics: Adenosine Triphosphate; Animals; Animals, Newborn; Brain; Brain Ischemia; Cell Membrane; Energy Metabolism; Glucose; Hyperglycemia; Hypoxia; Lactic Acid; Phosphocreatine; Sodium-Potassium-Exchanging ATPase; Spectroscopy, Near-Infrared; Swine

Acidosis may contribute to ischemic injury by mobilizing iron because the iron chelator deferoxamine improves early metabolic recovery from hyperglycermic ischemia. Mobilized iron may then promote oxygen radical-induced lipid peroxidative injury during reperfusion. We tested the hypothesis that administration of the antioxidant tirilazad at the start of reperfusion improves early metabolic recovery after severe acidotic ischemia and ameliorates depletion of the endogenous antioxidant glutathione.. In anesthetized dogs, arterial glucose concentration was increased to 500 to 600 mg/dL and global incomplete cerebral ischemia was produced for 30 minutes by ventricular fluid infusion to reduce perfusion pressure to 10 to 12 mm Hg. Metabolic recovery and intracellular pH were measured by phosphorus MR spectroscopy. In the first experiment, four groups of eight dogs each received either vehicle or 0.25, 1, or 2.5 mg/kg of tirilizad mesylate at reperfusion. Cerebral blood flow was measured with microspheres. In the second experiment, two groups of eight dogs each each received either vehicle or 2.5 mg/kg of tirilazad at reperfusion, and cortical glutathione was measured at 3 hours of reperfusion.. Cerebral blood flow decreased to approximately 6 mL/min per 100 g and intracellular pH decreased to approximately 5.6 during ischemia in all groups. In the vehicle group, ATP recovery was transient and pH remained less than 6.0. Cerebral blood flow, O2 consumption, and ATP eventually declined to near-zero levels by 3 hours. Recovery was improved by tirilazad posttreatment in a dose-dependent fashion. At the highest dose, cerebral blood flow and O2 consumption were sustained near preischemic levels, and five of eight dogs had recovery of ATP greater than 50% and of pH greater than 6.7. Recovery of ATP and phosphocreatine became significantly greater than that in the vehicle group by 17 minutes of reperfusion despite similar levels of early hyperemia, indicating that the drug was acting before the onset of hypoperfusion. Cortical glutathione concentration in the vehicle group was 27% less than that in the tirilazad group and 34% less than that in nonischemic controls.. Decreased depletion of the endogenous antioxidant glutathione is consistent with tirilazad acting as an antioxidant in vivo. Improvement in high-energy phosphate recovery 17 minutes after starting tirilazad infusion during reperfusion is consistent with an early onset of a functionally significant oxygen radical injury. Thus, severe acidosis appears to contribute to early ischemic injury through an oxygen radical mechanism sufficient to impede metabolic recovery.

Topics: Acidosis; Adenosine Triphosphate; Animals; Antioxidants; Brain; Brain Ischemia; Cerebrovascular Circulation; Dogs; Dose-Response Relationship, Drug; Free Radical Scavengers; Glutathione; Hydrogen-Ion Concentration; Hyperemia; Hyperglycemia; Iron; Lipid Peroxidation; Magnetic Resonance Spectroscopy; Male; Oxygen Consumption; Phosphocreatine; Phosphorus; Pregnatrienes; Reperfusion

Unlike adults, hyperglycemia with circulating glucose concentrations of 25-35 mM/L protects the immature brain from hypoxic-ischemic damage. To ascertain the effect of hyperglycemia on cerebral oxidative metabolism during the course of hypoxia-ischemia, 7-day postnatal rats underwent unilateral common carotid artery ligation followed by exposure to 8% O2 for 2 h at 37 degrees C. Experimental animals received 0.2 cc s.c. 50% glucose at the onset of hypoxia-ischemia, and 0.15 cc 25% glucose 1 h later to maintain blood glucose concentrations at 20-25 mM/L for 2 h. Control rat pups received equivalent concentrations or volumes of either mannitol or 1 N saline at the same intervals. The cerebral metabolic rate for glucose (CMRglc) increased from 7.1 (control) to 20.2 mumol 100 g-1 min-1 in hyperglycemic rats during the first hour of hypoxia-ischemia, 79 and 35% greater than the rates for saline-and mannitol-injected animals at the same interval, respectively (p < 0.01). Brain intracellular glucose concentrations were 5.2 and 3.0 mM/kg in the hyperglycemic rat pups at 1 and 2 h of hypoxia-ischemia, respectively; glucose levels were near negligible in mannitol- and saline-treated animals at the same intervals. Brain intracellular lactate concentrations averaged 13.4 and 23.3 mM/kg in hyperglycemic animals at 1 and 2 h of hypoxia-ischemia, respectively, more than twice the concentrations estimated for the saline- and mannitol-treated littermates. Phosphocreatine (PCr) and ATP decreased in all three experimental groups, but were preserved to the greatest extent in hyperglycemic animals. Results indicate that anaerobic glycolytic flux is increased to a greater extent in hyperglycemic immature rats than in normoglycemic littermates subjected to cerebral hypoxia-ischemia, and that the enhanced glycolysis leads to greater intracellular lactate accumulation. Despite cerebral lactosis, energy reserves were better preserved in hyperglycemic animals than in saline-treated controls, thus accounting for the greater resistance of hyperglycemic animals to hypoxic-ischemic brain damage.

Topics: Adenosine Triphosphate; Animals; Body Water; Brain; Brain Ischemia; Carotid Arteries; Glucose; Glycolysis; Hyperglycemia; Hypoxia, Brain; Kinetics; Lactates; Lactic Acid; Ligation; Oxygen; Phosphocreatine; Rats; Rats, Wistar

Sepsis and endotoxin (LPS) have been demonstrated to impair insulin-mediated glucose uptake in skeletal muscle. However, the intracellular mechanism responsible for this defect is not fully defined. The purpose of the present study was to determine whether specific elements of the insulin receptor (IR) signaling pathway in skeletal muscle are altered by LPS. In vivo injection of Escherichia coli LPS resulted in a 44% reduction in whole body glucose disposal under euglycemic hyperinsulinemic conditions, which was largely accounted for by a decreased rate of glycogen synthesis. Scatchard analysis indicated that the number and affinity of the high-affinity insulin binding sites in muscle were similar between control and LPS-treated rats. Western blot analysis indicated that under basal conditions, the levels of total and phosphorylated IR, insulin receptor substrate (IRS)-1, and mitogen-activated protein (MAP) kinase were not significantly different between control and endotoxic rats. In control animals, muscle obtained 2 min after intravenous injection of a maximally stimulating dose of insulin demonstrated a marked increase in the amount of phosphorylated IR (approximately 5-fold), IRS-1 (approximately 10-fold), and MAP kinase (approximately 10-fold). Insulin-stimulated phosphorylation of IR, IRS-1, and MAP kinase was markedly diminished (approximately 75%, 90%, and 78%, respectively) in LPS-treated rats. However, there was no concomitant reduction in the total abundance of these proteins under hyperinsulinemic conditions. These data demonstrate that LPS alters multiple steps in the insulin signal transduction pathway, but not insulin binding, in skeletal muscle that may mediate the observed impairment in glucose uptake.

Topics: Adenosine Triphosphate; Animals; Blood Glucose; Calcium-Calmodulin-Dependent Protein Kinases; Endotoxins; Glucose; Glycogen; Hyperglycemia; Insulin; Insulin Receptor Substrate Proteins; Lipopolysaccharides; Male; Muscle, Skeletal; Phosphocreatine; Phosphoproteins; Phosphorylation; Rats; Rats, Sprague-Dawley; Signal Transduction

Hyperglycemia increases cerebral damage after transient cerebral ischemia. This study used in vivo 31P nuclear magnetic resonance spectroscopy to determine the relationship of intracellular tissue acidosis and delayed recovery of brain high-energy phosphates to increased damage during the reperfusion period. Mongolian gerbils were subjected to transient bilateral carotid ischemia for 20 min with 2 h reperfusion. All gerbils were pretreated intraperitoneally with equivalent volumes in saline of 0.003 units per kilogram of insulin or vehicle, or with 4 grams of glucose per kilogram. The gerbils were then scanned in a 4.7 Tesla Magnetic Resonance Imager-Spectrometer to determine levels of intracellular pH, inorganic phosphate, adenosine triphosphate, and phosphocreatine. In each group, intracellular pH decreased with ischemia, but most significantly in hyperglycemic animals (6.45 +/- 0.15), in which it had not recovered to preischemic levels by the end of the reperfusion period (6.8 +/- 0.1 vs 7.04 +/- 0.1, p < 0.05). High-energy phosphates phosphocreatine-inorganic phosphate and phosphocreatine-adenosine triphosphate showed partial recovery in all groups throughout the reperfusion period; the recovery was not significantly altered by glucose status. Hyperglycemia worsened pH but not the recovery of high-energy phosphates in animals reperfused after 20 min of transient cerebral ischemia. This sustained acidosis may be a primary event in transient damage in hyperglycemic animals.

Topics: Adenosine Triphosphate; Animals; Arterial Occlusive Diseases; Blood Glucose; Body Weight; Brain Ischemia; Carotid Arteries; Energy Metabolism; Gerbillinae; Glucose; Hydrogen-Ion Concentration; Hyperglycemia; Hypoglycemic Agents; Insulin; Magnetic Resonance Spectroscopy; Male; Phosphates; Phosphocreatine; Reperfusion Injury

We used 31P nuclear magnetic resonance (NMR) spectroscopy to study the effect of moderate hyperglycemia on brain ATP and intracellular pH in a model of severe incomplete forebrain ischemia. Plasma glucose in the hyperglycemic rats was 277 +/- 9 mg/100 ml compared with 115 +/- 10 mg/100 ml in the normoglycemic rats at the onset of ischemia. After 15 min of ischemia, brain ATP levels decreased to 31 +/- 8% in normoglycemic rats vs. 63 +/- 11% in hyperglycemic rats (P < 0.05). Phosphocreatine levels were 31 +/- 9 and 55 +/- 8% for normoglycemic and hyperglycemic rats, respectively. Intracellular pH decreased to the same level (approximately 6.5) in both normoglycemic and hyperglycemic animals after 15 min of ischemia. In summary, we found that moderate hyperglycemia during severe incomplete forebrain ischemia significantly increases ischemic brain ATP levels but does not have a significant effect on intracellular pH. These results support the hypothesis that alterations in brain ATP and adenosine concentrations may be important in the pathogenesis of ischemic tissue injury under moderate hyperglycemic conditions, whereas alterations in tissue pH may be less important.

Topics: Adenosine Triphosphate; Animals; Brain; Brain Ischemia; Hydrogen-Ion Concentration; Hyperglycemia; Intracellular Membranes; Magnetic Resonance Spectroscopy; Male; Phosphocreatine; Phosphorus; Rats; Rats, Sprague-Dawley

31-phosphorous magnetic resonance spectroscopy was used in a rat model of 10 min severe incomplete forebrain ischemia (two-vessel occlusion with hypotension) to assess the effect of hyperglycemia on intracellular pH and high energy phosphates during ischemia and early reperfusion. One group (n = 8) with preischemic hyperglycemia (serum glucose 20 mmol.l-1) showed an increased intracellular acidosis (pH 6.35) during ischemia compared to 6.55 in the normoglycemic control group (n = 7, P less than 0.001), but the recovery of phosphocreatine and ATP in early reperfusion was the same in the two groups. Another group (n = 7) was normoglycemic during ischemia, but received an i.v. bolus of glucose during the first minute of reperfusion. In this group the recovery of intracellular pH in early reperfusion was slower than in the control group (0.034 +/- 0.006 pH units per minute compared to 0.052 +/- 0.11 in the controls, +/- s.d. and P less than 0.01).

Topics: Acidosis; Adenosine Triphosphate; Animals; Blood Glucose; Brain Ischemia; Hydrogen-Ion Concentration; Hyperglycemia; Magnetic Resonance Spectroscopy; Male; Phosphates; Phosphocreatine; Phosphorus; Rats; Rats, Inbred Strains; Reperfusion; Time Factors

This study investigated the relations between hemorrhagic infarction and occlusion, release, levels of glycemia, brain energy state, and lactate content after cerebrovascular occlusion.. Prospective, controlled laboratory investigation.. One hundred six pentobarbital-anesthetized cats.. The middle cerebral artery was occluded with a Yasargil clip transorbitally either temporarily (0.5, four, and eight hours) or permanently. Normoglycemic and hyperglycemic animals were closely monitored for eight hours. Brain pathology was assessed after two weeks' survival or at the time of spontaneous animal death. Topographic brain metabolite studies were carried out after four hours of middle cerebral artery occlusion.. Morphometric quantitation of cerebral hemorrhage and infarction and fluorometric determinations of blood and brain tissue, glucose, glycogen, lactate, adenosine triphosphate, and phosphocreatine from 16 topographic brain sites were carried out. Twenty-one of 82 (25.6%) animals evaluated neuropathologically showed hemorrhagic infarcts. Occluding the artery in hyperglycemic animals caused fivefold more frequent and 25-fold more extensive hemorrhage into infarcts than in normoglycemic animals. Temporary occlusion with clip release after four hours in hyperglycemic animals caused the most extensive hemorrhage into infarcts. Most hemorrhages into infarcts (81%) took place in animals that died within a few hours after they experienced ischemia and that showed infarction and marked edema of the entire middle cerebral artery territory. Linear regression analyses demonstrated a close relation between hemorrhage into infarcts and near-total energy depletion (adenosine triphosphate, less than 0.3 microM/g; phosphocreatine, less than 0.5 microM/g) in brain sites that showed extremely high tissue lactate concentrations (more than 30 microM/g). The biochemical changes that correlated with hemorrhage into infarcts were more marked than those with infarcts without hemorrhage.. Hyperglycemia and restoration of blood flow to ischemic territories were strong risk factors for hemorrhagic infarct conversion. Concomitant tissue metabolic changes suggest that marked tissue energy depletion accompanied by acidosis damages brain vessels and renders them penetrable for edema fluid and, ultimately, red blood cell extravasation.

Topics: Adenosine Triphosphate; Animals; Brain; Brain Chemistry; Brain Ischemia; Cats; Cerebral Arteries; Cerebral Hemorrhage; Cerebral Infarction; Constriction; Glucose; Hyperglycemia; Lactates; Lactic Acid; Linear Models; Phosphocreatine; Prospective Studies; Risk Factors

Brain oedema is an important aspect of infarction from cerebrovascular occlusion. In a cat stroke model where the middle cerebral artery (MCA) was reversibly or permanently occluded, we analyzed the incidence of fatal hemispheral oedema in 35 normo- (6 mM) and 35 hyperglycaemic (20 mM for 6 hours) animals, with (N = 45) and without (N = 25) restoration of blood flow with clip release at 4 and 8 hrs of occlusion. Fatal hemispheral oedema occurred in 23% of cats (16/70) while hyperglycaemia, for one, and restoration of blood flow, for another, each quadrupled its occurrence. Further, evidence of remote oedema in the form of posterior cingulate cortical pressure atrophy from transtentorial herniation was found in animals that were allowed to survive for 2 weeks and that exhibited infarcts that affected 12 to 95% of the MCA territory. Thus, hemispheral oedema in association with MCA occlusion developed sufficiently markedly as to cause transtentorial herniation in 47% of all cats (33/70). We carried out biochemical analyses in 14 hyper- and 10 normoglycaemic cats after 4 hrs of MCA occlusion for ATP, phosphocreatine (PCr), lactate, glucose and glycogen. The biochemical findings then were correlated with the occurrence of reperfusion oedema following clip release after 4 hrs of occlusion point-by-point in the brains. Linear regression analyses of the brain metabolic and pathologic data revealed highly significant (p less than 0.001) correlations of acute oedema with brain tissue ATP and PCr reductions less than 1.5 microM/g, with lactic acid accumulation greater than 20 microM/g and with the extents of reduction in brain tissue glucose concentrations in the ischaemic territories.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenosine Triphosphate; Animals; Brain; Brain Edema; Brain Ischemia; Cats; Forecasting; Hyperglycemia; Phosphocreatine; Regression Analysis; Reperfusion

Progressive cerebral ischemia was induced in seven anesthetized hyperglycemic rats by carotid artery ligation and hemorrhagic hypotension. Phosphorus metabolites, intracellular pH, and lactate in the brain were monitored by 31P and 1H magnetic resonance spectroscopy. Under conditions in which blood flow was low, phosphocreatine (PCr) concentration and intracellular pH decreased and the concentration of lactate increased. The decrease in ATP was approximately one-third that of PCr until only 25% PCr remained, after which ATP was lost more rapidly than PCr. These changes were interpreted in terms of three regions observed by the magnetic resonance coil, one of complete ischemia, one of partial ischemia, and one of perfusion sufficient to maintain normal metabolite levels. The extent of the three regions was estimated quantitatively. Broadening and splitting of the inorganic phosphorus (Pi) peak into two components provided further evidence of distinct populations of cells, one very acidic and another less so. Apparent intracellular buffering capacity was calculated as 23.6 +/- 1.3 mumol lactate/g wet wt/pH.

Topics: Acidosis, Lactic; Adenosine Triphosphate; Animals; Brain Ischemia; Hydrogen; Hydrogen-Ion Concentration; Hyperglycemia; Lactates; Magnetic Resonance Spectroscopy; Phosphocreatine; Phosphorus; Rats; Rats, Inbred Strains

Unlike adult rats, glucose supplementation of immature rats does not lead to accentuated hypoxic-ischemic brain damage. To explore the reason for this age-specific paradox, we subjected 7-day postnatal rats to unilateral common carotid artery occlusion followed by a subcutaneous injection of either 0.1 ml 50% glucose or normal saline. They were then exposed to hypoxia with 8% oxygen, during which they received 2.5 microCi 2-[14C]-glucose or were quick-frozen for brain metabolite analysis. During hypoxia-ischemia, glucose transport into the ipsilateral cerebral hemisphere of the hyperglycemic rats was greater (+100-150%) than in normoglycemic animals. However, glucose consumption was similar in the two groups. Glucose concentrations in brain were lower during hypoxia-ischemia in the normoglycemic animals, whereas lactate increased to similar levels in the two groups. The high-energy phosphate reserves, ATP and phosphocreatine, were depleted to a similar extent. Thus, hyperglycemia combined with hypoxia-ischemia, although associated with increased glucose transport into brain, does not lead to enhanced glucose utilization or lactate accumulation by brain over that of hypoxia-ischemia alone.

Topics: Adenine Nucleotides; Animals; Animals, Newborn; Blood Glucose; Brain; Brain Ischemia; Glucose; Hyperglycemia; Hypoxia, Brain; Lactates; Lactic Acid; Phosphocreatine; Pyruvates; Pyruvic Acid; Rats; Rats, Inbred Strains

The hyperglycemia-induced in vivo metabolic changes produced in subcutaneous murine RIF-1 tumors, grown on female C3H/Anf mice, were examined with 31P surface-coil NMR. Serum glucose levels were elevated 4-fold by bolus intraperitoneal injection of 0.3 ml of an aqueous 50% glucose solution. Tumor pH was calculated from the chemical shift of Pi and relative phosphocreatine and ATP concentrations were determined by Simpson's rule integration of the peak areas. Tumor pH decreased by ca. 0.45 unit over 2 hr while phosphocreatine concentrations decreased by ca. 50% over the same time period (n = 9). Initial tumor pH correlated inversely with the initial peak intensity ratio of Pi:ATP (r = -0.77). In a significant number of tumors (n = 4), two pH populations were observed. In these tumors, one population was unaffected by hyperglycemia and the other showed a decrease in pH. In the other tumors (n = 5), the pH distribution broadened as the pH decreased. In these tumors, the observed decreased in phosphocreatine concentration correlated with that calculated from the effect of measured tumor pH on the intracellular creatine kinase equilibrium (n = 18, r = 0.91). This correlation and consideration of the Pi distribution in the tumor suggest that the pH measured by 31P NMR is weighted heavily by intracellular pH for the RIF-1 tumor. The presence of two distinct tumor pH populations or a broadened pH distribution likely reflects variations in tumor microcellular environment. Control experiments showed negligible changes in tumor pH and high energy phosphate concentrations after bolus intraperitoneal injection of 0.3 ml of isotonic saline. In addition, negligible changes in leg muscle pH and high energy phosphate concentrations were observed after glucose injection into mice with or without tumors. These results indicate that hyperglycemia induced by intraperitoneal glucose injection is effective in lowering the tumor pH of the murine RIF-1 tumor.

Topics: Adenosine Diphosphate; Adenosine Triphosphate; Animals; Fibrosarcoma; Hydrogen-Ion Concentration; Hyperglycemia; Magnetic Resonance Spectroscopy; Mice; Mice, Inbred C3H; Phosphates; Phosphocreatine; Sarcoma, Experimental

An hypothesis is presented relating several well-defined metabolic abnormalities in diabetic peripheral nerve to impaired peripheral nerve function by a sodium-potassium ATPase mechanism. It is proposed that this hypothesis be tested in the most well-defined animal model for human insulin deficiency diabetes currently available--the BB diabetic rat.

Topics: Animals; Diabetes Mellitus, Experimental; Diabetes Mellitus, Type 1; Energy Metabolism; Female; Glucose; Hyperglycemia; Inositol; Male; Peripheral Nerves; Phosphocreatine; Rats; Rats, Inbred Strains; Sodium-Potassium-Exchanging ATPase

Topics: Adenosine Triphosphate; Animals; Brain Chemistry; Hyperglycemia; Male; Muscles; Myocardium; Phosphocreatine; Rats

Topics: Acidosis; Adenosine Diphosphate; Adenosine Monophosphate; Adenosine Triphosphate; Animals; Blood Glucose; Blood Pressure; Body Temperature; Brain; Creatine; Energy Metabolism; Glucosephosphates; Glycogen; Hydrogen-Ion Concentration; Hyperglycemia; Hypoglycemia; Ischemia; Lactates; Phosphocreatine; Pyruvates; Rats