Optimizing suture middle cerebral artery occlusion model in C57BL/6 mice circumvents posterior communicating artery dysplasia

The suture middle cerebral artery occlusion (MCAO) model is used worldwide in both academia and industry. However, the variable occurrence of dysplasia in posterior communicating arteries (PcomAs) induces high mortality and instability in permanent MCAO models, limiting the model's application to transient focal ischemia. In particular, high mortality in intraluminal suture MCAO models is associated with the dysplasia of PcomAs in C57BL/6 mice. Optimization of silicone coating length is critical for reducing mortality and generating stable infarct in this model. The aim of our study is to reduce mortality and improve the reproducibility of the intraluminal suture MCAO model in C57BL/6 mice, which have high variation in PcomA dysplasia. Adult male C57BL/6 mice (n=38) underwent MCAO using sutures with various diameters and silicone coating lengths. The occlusion of cerebral vessels was examined by synchrotron radiation live angiography. The morphology of PcomAs was examined under a microscope after MICROFIL(?) infusion. Neurological outcome, infarct volume, and mortality were examined within 28 days. Optimizing the silicone coating on an 8-0 suture tip, we were able to reduce the model mortality to zero after permanent occlusion in C57BL/6 and produce stable brain infarct volume independent of the patency of PcomAs.     

Beta-adrenoreceptor antagonists attenuate brain injury after transient focal ischemia in rats

Beta-adrenoreceptor antagonists experimentally reduce cardiac and renal injury after ischemia and are also clinically useful for myocardial infarction and severe burns. In addition, beta-adrenoreceptor antagonists provide neuroprotective effects after focal cerebral ischemia in experimental settings. We conducted the present study to compare the neuroprotective effects of several beta-adrenoreceptor antagonists in rat transient focal cerebral ischemia. Halothane-anesthetized normothermic adult male Sprague-Dawley rats were subjected to 2 h of middle cerebral artery occlusion using the intraluminal suture technique confirmed by laser Doppler flowmetry. Rats received an IV infusion of saline 0.5 mL/h, propranolol 100 microg x kg(-1) x min(-1), carvedilol 4 microg x kg(-1) x min(-1), esmolol 200 microg x kg(-1) x min(-1), or landiolol 50 microg x kg(-1) x min(-1) (n = 6 in each group). Infusion was initiated 30 min before middle cerebral artery occlusion and continued for 24 h. Additional rats received esmolol 50 microg x kg(-1) x min(-1) or landiolol 10 microg x kg(-1) x min(-1) intrathecally (IT) via the cisterna magna (n = 5 in each group), according to the same experimental protocol. The neurological deficit score was evaluated at 22 h after reperfusion, and the brains were removed and stained with triphenyltetrazolium chloride for evaluation of infarct volume. Additional rats that received saline, esmolol, and landiolol IV (n = 6 in each group) were allowed to survive for 7 days followed by measurement of infarct size. Neurological deficit scores were smaller in rats treated with propranolol-IV, carvedilol-IV, esmolol-IV, landiolol-IV, esmolol-IT, and landiolol-IT compared with saline-treated rats (P < 0.05). Cortical and striatum infarct volumes were less in the rats receiving beta-adrenoreceptor antagonists via either IV or IT than in saline-treated rats (P < 0.05). We conclude that beta-adrenoreceptor antagonists improve neurological and histological outcomes after transient focal cerebral ischemia in rats independent of administration route.     

Protective Effects of Memantine Against Ischemia-Reperfusion Injury in Spontaneously Hypertensive Rats

Summary  Memantine, an uncompetitive NMDA open-channel blocker, has been shown to be effective in preventing neuronal damage after permanent focal cerebral ischemia. Reperfusion after a long period of ischemia may aggravate the progression of neuronal damage. Those drugs that show protective effects after permanent cerebral ischemia, therefore, might fail to do so against ischemia-reperfusion injury. In this study we evaluated the effects of memantine on brain edema formation and ischemic injury volume after transient cerebral ischemia. Male Spontaneously Hypertensive Rats (SHR) weighing 250–300 g were anesthetized with halothane and subjected to 1 hour of temporary middle cerebral artery occlusion by an intraluminal suture. 20 mg/kg of memantine or saline were injected intraperitoneally 5 min. after the induction of ischemia. Physiological parameters and regional cerebral blood flow were monitored during the surgical procedure. Brain water content and ischemic injury volume were measured with the wet dry method and 2,3,5-triphenyl tetrazolium chloride monohydrate (TTC) staining, respectively, at 24 hours after occlusion. There were no statistically significant differences between the groups regarding physiological parameters during the procedure. Memantine treatment (n=9) reduced the brain water content significantly in the cortex compared to saline treatment (n=8; 83.1±0.7% vs. 84.5±1.5%, respectively, p<0.05). The total volume of ischemic brain injury was 300±49 mm³ in the animals treated with saline (n=13). Treatment with 20 mg/kg memantine (n=14) reduced the ischemic injury volume to 233±61 mm³ (P<0.01). These results demonstrate that the harmful effects of recirculation after a period of ischemia can be attenuated by the treatment of memantine, perhaps by its action at the NMDA receptors.     

丙泊酚预处理减轻大鼠短暂性局部脑缺血再灌注后的脑水肿及AQP4的过度表达

背景脑水肿是脑卒中患者面临的主要威胁。许多研究关注丙泊酚的神经保护作用,多着重于其减少梗死面积而非降低脑水肿方面。大脑经受各种神经损伤后,在维持脑内水环境稳定方面水通道蛋白-4（AQP4）发挥了重要作用。我们探讨了在大鼠脑缺血再灌注模型中丙泊酚预处理对脑水肿的作用,并评估了AQP4所起的作用。方法为了建立舾缺血再灌注模型,我们用涂抹硅胶的单丝尼龙线阻塞大脑中动脉起始处,90分钟后拔线。处理组（n=32）在阻塞前30分钟持续泵注丙泊酚0．1m1·kg^-1·min^-1,对照组（n=32）及假手术组（n=28）在相同时间以同样的速率泵注脂肪乳。用2,3,5．氯化三苯四唑染色法测定脑梗死体积;在湿．千重比率判断脑水肿程度的基础上,免疫组织化学法及蛋白印迹法检测AQP4表达。结果湿一干重比率在对照组（n=6）为86．89％±0．71％,丙泊酚组（n=6）降至72．42％±0．74％,平均下降16％。免疫组织化学法半定量分析显示,丙泊酚组（n=7）缺血边缘区的AQP4过度表达较对照组（n=7）显著降低,分别为1．28±0．03和1．40±0．05,P〈0．05。蛋白印迹定量同样显示丙泊酚组（n=4）较对照组（n=4）下降：分别为20．85％±4．18％和31．67％±3．23％,P〈0．05。然而,梗死体积及缺血后大鼠神经功能缺失程度在丙泊酚组和对照组间并没有显著统计学差异。结论丙泊酚预处理能够减轻大鼠缺血后脑水肿程度,可能的机制是降低了缺血边缘区AQP4的过度表达。     

Comparative neuroprotective efficacy of prolonged moderate intraischemic and postischemic hypothermia in focal cerebral ischemia

OBJECT: The purpose of this study was to compare the effects of prolonged hypothermia on ischemic injury in a highly reproducible model of middle cerebral artery (MCA) occlusion in rats. METHODS: Male Sprague-Dawley rats were anesthetized with halothane and subjected to 120 minutes of temporary MCA occlusion by retrograde insertion of an intraluminal nylon suture coated with poly-L-lysine through the external carotid artery into the internal carotid artery and the MCA. Two levels of prolonged postischemic cranial hypothermia (32 degrees C and 27 degrees C) and one level of intraischemic cranial hypothermia (32 degrees C) were compared with the ischemic normothermia (37 degrees C) condition. Target cranial temperatures were maintained for 3 hours and then gradually restored to 35 degrees C over an additional 2-hour period. The animals were evaluated using a quantitative neurobehavioral battery of tests before inducing MCA occlusion, during occlusion (at 60 minutes postonset in all rats except those in the intraischemic hypothermia group), and at 24, 48, and 72 hours after reperfusion. The rat brains were perfusion fixed at 72 hours after ischemia, and infarct volumes and brain edema were determined. Both intraischemic and postischemic cooling to 32 degrees C led to similar significant reductions in cortical infarct volume (by 89% and 88%, respectively) and total infarct volume (by 54% and 69%, respectively), whereas postischemic cooling to 27 degrees C produced lesser reductions (64% and 49%, respectively), which were not statistically significant. All three hypothermic regimens significantly lessened hemispheric swelling and improved the neurological score at 24 hours. The authors' data confirm that a high degree of histological neuroprotection is conferred by postischemic cooling to 32 degrees C, which is virtually equivalent to that observed with intraischemic cooling to the same level. CONCLUSIONS: These results may be relevant to the design of future clinical trials of therapeutic hypothermia for acute ischemic stroke.     

Effect of mannitol on local cerebral blood flow after temporary complete cerebral ischemia in rats.

The effects of pretreatment with mannitol on local cerebral blood flow (CBF) after permanent or temporary global cerebral ischemia were evaluated with 14C-iodoantipyrine autoradiography in rats under halothane-N2O endotracheal anesthesia. Blood pressure, pulse rate, arterial blood gas levels, and electroencephalographic (EEG) tracings were monitored throughout the experiments. After permanent occlusion of the basilar artery and both external carotid and pterygopalatine arteries, severe global ischemia was induced by permanent occlusion of the common carotid arteries (CCA's) or by a 30-minute temporary CCA occlusion followed by 5 minutes of reperfusion. Intravenous mannitol (25%, 1 gm/kg) or saline solution was administered 5 minutes before occlusion of the CCA's. Cerebral blood flow was measured in 24 anatomical regions. The EEG tracings flattened within 2 to 3 minutes after the onset of ischemia, and no recovery was observed during reperfusion. In the mannitol-treated rats and the saline-treated controls, autoradiographic studies after permanent occlusion showed no CBF in the forebrain or cerebellum, although brain-stem and spinal cord CBF values were normal. After 5 minutes of reperfusion, CBF in the cortex, basal ganglia, and white matter was 100% to 200% higher in mannitol-treated rats and 50% to 100% higher in saline-injected rats than in the nonischemic anesthetized control group. Heterogeneously distributed areas of no-reflow were seen in all saline-injected rats but were observed in none of the mannitol-treated rats. Pretreatment with mannitol prevented postischemic obstruction of the microcirculation during 5 minutes of recirculation after 30 minutes of severe temporary ischemia, but the EEG signals did not recover. Further studies of the functional and morphological responses to longer periods of postischemic recirculation are needed to verify the extent to which these mannitol-induced effects are protective.     

Magnesium sulfate and nimesulide have synergistic effects on rescuing brain damage after transient focal ischemia

Magnesium sulfate and nimesulide are commonly used drugs with reported neuroprotective effects. Their combination as stroke treatment has the potential benefits of decreasing individual drug dosage and fewer adverse effects. This study evaluated their synergistic effects and compared a low-dose combination with individual drug alone and placebo. Sprague-Dawley rats underwent 90?min of focal ischemia with intraluminal suture occlusion of the middle cerebral artery followed by reperfusion. The rats were randomly assigned to receive one of the following treatments: placebo, magnesium sulfate (MgSO?; 45?mg/kg) intravenously immediately after the induction of middle cerebral artery occlusion, nimesulide (6?mg/kg) intraperitoneally before reperfusion, and combined therapy. Three days after the ischemia-reperfusion insult, therapeutic outcome was assessed by 2,3,5-triphenyltetrazolium chloride staining and a 28-point neurological severity scoring system. Cyclooxygenase-2, prostaglandin E?, myeloperoxidase, and caspase-3 expression after treatment were evaluated using Western blot analyses and immunohistochemical staining, followed by immunoreactive cell analysis using tissue cytometry. Only the combination treatment group showed a significant decrease in infarction volume (10.93±6.54% versus 26.43±7.08%, p<0.01), and neurological severity score (p<0.05). Low-dose MgSO? or nimesulide showed no significant neuroprotection. There was also significant suppression of cyclooxygenase-2, prostaglandin E?, myeloperoxidase, and caspase-3 expression in the combination treatment group, suggesting that the combination of the two drugs improved the neuroprotective effects of each individual drug. MgSO? and nimesulide have synergistic effects on ischemia-reperfusion insults. Their combination helps decrease drug dosage and adverse effects. Combined treatment strategies may help to combat stroke-induced brain damage in the future.     

The effect of prior elevation of skin flaps and ischemia on blood thromboxane levels

The physiological factors that allow for the survival of ischemic skin flaps have not been clearly elucidated. Previous work by others has shown that elevation of a flap 24 hours before an episode of complete ischemia significantly improved survival, presumably by delaying the onset of the no-reflow phenomenon. The current study, using an epigastric flap, investigated the role of thromboxane in these events by observing postischemic plasma levels of thromboxane B2, the stable metabolite of the short-lived thromboxane A2. Acutely ischemic flaps were compared with those elevated 24 hours before ischemia. After the ischemic insult, blood was drawn from the venous effluent of the flaps. Thromboxane levels after 4 hours of ischemia were significantly decreased (p less than 0.001) in the postischemic period in those flaps elevated 24 hours before ischemia compared with flaps that had undergone ischemia acutely. Moreover, acutely ischemic flaps had significantly more thromboxane than nonischemic controls (p less than 0.001). These results confirm the importance of thromboxane metabolism in the no-reflow phenomenon.     

Superiority of early relative to late ischemic preconditioning in spinal cord protection after descending thoracic aortic occlusion

OBJECTIVE: We previously showed that ischemic preconditioning significantly reduced spinal cord injury caused by 35-minute aortic occlusion. In this study we investigated the effect of ischemic preconditioning on spinal cord injury after 45-minute aortic occlusion. METHODS: Thirty-two pigs were divided as follows: group 1 (n = 6) underwent sham operation, group 2 (n = 6) underwent 20 minutes of aortic occlusion, group 3 (n = 6) underwent 45 minutes of occlusion, group 4 (n = 6) underwent 20 minutes of occlusion and 48 hours later underwent an additional 45 minutes, and group 5 (n = 8) underwent 20 minutes of occlusion and 80 minutes later underwent an additional 45 minutes. Aortic occlusion was accomplished with two balloon occlusion catheters placed fluoroscopically after the origin of the left subclavian artery and at the aortic bifurcation. Neurologic evaluation was by Tarlov score. The lower thoracic and lumbar spinal cords were harvested at 120 hours and examined histologically with hematoxylin-eosin staining. The number of neurons was counted, and the inflammation was scored (0-4). Statistical analysis was by Kruskal-Wallis and 1-way analysis of variance tests. RESULTS: Group 5 (early ischemic preconditioning) had better Tarlov scores than group 3 ( P < .001) and group 4 (late ischemic preconditioning, P < .001). The histologic changes were proportional to the Tarlov scores, with the least histologic damage in the animals of group 5 relative to group 3 (number of neurons P < .001, inflammation P = .004) and group 4 (number of neurons P < .001, inflammation P = .006). CONCLUSION: Early ischemic preconditioning is superior to late ischemic preconditioning in reducing spinal cord injury caused by the extreme ischemia of 45 minutes of descending thoracic aortic occlusion.     

Monoamine metabolism after transient global ischemia. Mechanism of delayed postischemic hypoperfusion

During the period of recirculation following transient global ischemia, an initial hyperemia is succeeded by a secondary decrease in cerebral blood flow, termed "delayed postischemic hypoperfusion." It has been suggested that this phenomenon can lead to additional brain damage after the initial ischemic insult. One proposed mechanism of delayed postischemic hypoperfusion is increased cerebrovascular smooth muscle tone. Release of vasoactive amines, formation of vasoactive products of arachidonic acid metabolism, and disturbance of calcium ion homeostasis in cerebrovascular smooth muscle may contribute to postischemic vasoconstriction. In this study, monoamine metabolism following transient global ischemia was investigate. Mongolian gerbils subjected to 15 minutes of temporal bilateral common carotid artery occlusion and up to 6 hours of recirculation were employed as a model of transient global ischemia. In this model, secondary energy failure reportedly occurs after 6 hours of recirculation. Regional cerebral concentrations of monoamines and their metabolites were determined by high-performance liquid chromatography with electrochemical detection. After 4 to 6 hours of recirculation, accumulation of vasoactive amine, 5-hydroxytryptamine, its major metabolite, 5-hydroxyindole acetic acid, and its precursor amino acid, tryptophan were detected. This finding strongly suggests a postischemic increase in both synthesis and release of this amine, which may explain postischemic vasoconstriction. Moreover, increased dopamine synthesis and release after 1 hour of recirculation was suggested. As dopamine release is reported to increase cerebral glucose utilization, its elevation may contribute to an increase in cerebral energy demand after ischemia. Thus, the brain becomes relatively ischemic and secondary ischemic cell damage occurs.     

Ischemic preconditioning of skeletal muscle improves tissue oxygenation during reperfusion

Ischemic preconditioning (IPC) renders tissue resistant to the deleterious effects of prolonged ischemia and reperfusion by prior exposure to brief, repeated periods of vascular occlusion. Although the mechanism by which IPC exerts its effect is unclear, it likely mediates an attenuation in capillary no-reflow. Tissue oximetry provides a potential technique to assess microvascular flow during ischemia/reperfusion and to measure the effect of IPC on muscle tissue oxygenation. The authors aimed to (a) establish that tissue oximetry is a sensitive method to assess the "no-reflow" phenomenon in skeletal muscle; and (b) to test the hypothesis that IPC would increase tissue oxygenation during reperfusion. In Group 1 (n = 5), the rabbit rectus femoris muscle was subjected to 2-hr ischemia. In Group 2 (n = 5), the muscle was subjected to 3.5-hr ischemia. In Group 3 (n = 6) the muscle was subjected to 3.5-hr ischemia preceded by three cycles of 10 min of pedicle occlusion and 10 min of reperfusion. Muscle oxygen tension was continuously monitored during the ischemic interval and for 6 hr of reperfusion. It was found that muscle oxygen tension in the flap at 5, 10, 30, 60, and 360 min after reperfusion was significantly decreased after 3.5-hr ischemia, compared with 2-hr ischemia (p < 0.05). Muscle oxygen tension at 30 and 60 min after reperfusion was significantly improved in the preconditioned group (p < 0.05). The results suggest that tissue oximetry is a sensitive method to assess tissue perfusion in reperfused skeletal muscle. Ischemic preconditioning improves tissue oxygenation during reperfusion following prolonged ischemia, which likely reflects an attenuation in capillary no-reflow.     

缺血预处理对家兔脑缺血保护效应的实验研究

目的 探讨缺血预处理脑保护效应,以及神经细胞凋亡与缺血性脑损害的关系。方法 家兔１５只,随机分为３组,对照组,缺血组,缺血预处理组。Ａ组只做手术操作,Ｂ组采用二血管夹闭全脑缺血１０分钟,Ｃ组在缺血前增加缺血预处理２分钟再灌注３０分钟。对比观察缺血后３天海马ＣＡ１区神经元密度和缺血细胞数,同时使用ＴＵＮＥＬ原位标记法,检测缺血３天后海马区的凋亡细胞。     

Potent sigma 1-receptor ligand 4-phenyl-1-(4-phenylbutyl) piperidine provides ischemic neuroprotection without altering dopamine accumulation in vivo in rats

The in vivo signaling of ischemic neuroprotection provided by sigma-receptor ligands remains unclear. Catecholamines have been implicated in the propagation of ischemic neuronal injury, and previous in vitro studies suggest that sigma ligands modulate dopaminergic neurotransmission. In this study, we tested the hypothesis that the potent sigma(1)-receptor ligand 4-phenyl-1-(4-phenylbutyl) piperidine (PPBP) attenuates the increase of extracellular dopamine in ischemic striatum. Under controlled physiological conditions, a microdialysis probe was implanted in right caudoputamen (CP) complex of adult male Wistar rats. Rats were subjected to 2 h of transient middle cerebral artery occlusion (MCAO) by the intraluminal suture technique. In a blinded, randomized fashion, rats were divided into five treatment groups: Group 1 (n = 8; saline-saline) continuous i.v. infusion of saline vehicle 30 min before MCAO followed by saline at reperfusion until the end of the experiment; Group 2 (n = 8; PPBP-PPBP) i.v. PPBP 30 min before MCAO followed by 1 micromol x kg(-1) x h(-1) of PPBP; Group 3 (n = 8; saline-PPBP) i.v. saline before MCAO followed by PPBP; Group 4 (n = 4) surgical shams (saline-saline); and Group 5 (n = 4) surgical shams (PPBP-PPBP). Infarction volume at 22 h of reperfusion in the CP complex (percentage of ipsilateral structure) was significantly attenuated in rats treated with PPBP-PPBP (27.3% +/- 9.1%) and saline-PPBP (27.8% +/- 12.7%) compared with saline-saline (59.3% +/- 7.3%) treatment. There was a three- to fourfold increase in dopamine concentrations in the microdialysates within 40 min of the onset of MCAO. Dopamine and its metabolites dihydroxy phenylacetic acid and homovallinic acid levels were similar among the three groups subjected to MCAO. Therefore, PPBP provides significant ischemic neuroprotection in the CP complex without altering the acute accumulation of dopamine in vivo during transient focal ischemia in the rat.     

Overexpression of rat heat shock protein 70 reduces neuronal injury after transient focal ischemia, transient global ischemia, or kainic acid-induced seizures

OBJECTIVE: Transgenic (Tg) mice overexpressing rat heat shock protein 70 (hsp70) demonstrated less infarction than did wild-type (WT) littermates after permanent focal cerebral ischemia. The purpose of this study was to determine whether neuronal injury and apoptosis were reduced in hsp70 Tg mice after transient focal ischemia. The effects of hsp70 overexpression were also evaluated after transient global ischemia or kainic acid (KA)-induced seizures, to verify the results in other excitotoxic stress models. METHODS: Transient focal ischemia was produced with middle cerebral artery occlusion via intraluminal suture cannulation. Infarction volumes were assessed 24 hours after 30 minutes of middle cerebral artery occlusion. Transient global ischemia was produced with 25 minutes of bilateral common carotid artery occlusion. KA (30 mg/kg) was administered subcutaneously, and seizure activity was evaluated. The number of eosinophilic neurons was assessed in the CA1 region 72 hours after bilateral common carotid artery occlusion and in the CA3 region 24 hours after KA administration. RESULTS: The infarction volume after transient middle cerebral artery occlusion was significantly smaller in hsp70 Tg mice than in WT mice (9.1 +/- 5.7 mm(3) versus 22.4 +/- 16.8 mm(3), P < 0.05). The number of eosinophilic neurons in the CA1 area after bilateral common carotid artery occlusion and in CA3 after KA injection was significantly lower in hsp70 Tg mice than in WT mice (949.1 +/- 1095.5 versus 2406.9 +/- 1380.3, P < 0.05, and 33.8 +/- 45.3 versus 119.4 +/- 112.1, P < 0.05, respectively). Fewer terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end-labeling-positive cells were observed in hsp70 Tg mice than in WT mice in each model. CONCLUSION: The results demonstrate that overexpression of hsp70 reduces neuronal injury after ischemia and seizures. The reduction in the number of terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end-labeling-positive cells in hsp70 Tg mice suggests that hsp70 overexpression might reduce apoptotic cell death.     

Intermittent ischemia potentiates intestinal reperfusion injury

We hypothesized that even brief periods of reperfusion interjected between ischemic episodes would increase tissue injury. Studies were performed in a rat small intestine preparation in which metabolic, hemodynamic, and histologic responses to ischemia have been well characterized. Animals were subjected to a total of 30 or 45 minutes of complete intestinal ischemia. Flow interruption was continuous (C, single episode) or intermittent (I, two or three episodes of 15-minute ischemia separated by 5 minutes of reperfusion). In some experiments 5-minute reperfusions were performed with arterial blood depleted of leukocytes (IL). This additional perturbation was included to determine the role of neutrophils that have been strongly implicated in reperfusion injury. In all three protocols histologic sections were obtained after each ischemic insult and after 1 hour of reperfusion with arterial blood. Villous histology was graded in a blinded fashion with 1 = normal and 5 = severe injury. No significant differences were found between groups in immediate postischemic histologies before reperfusion. After 1 hour of reperfusion, intermittent episodes of ischemia were associated with significantly worse histologic injury than that seen with comparable durations of continuous ischemia (30 min: I, 4.4 +/- 0.5 vs C, 2.7 +/- 0.4; 45 min: I, 4.9 +/- 0.2 vs C, 2.8 +/- 0.3). However, if 5-minute reperfusions were with leukopenic blood, this effect was markedly reduced (30 min IL, 3.4 +/- 0.3; 45 min IL, 3.6 +/- 0.2). Even short periods of reperfusion during an ischemic insult greatly increased mucosal injury.(ABSTRACT TRUNCATED AT 250 WORDS)     

Influence of Propofol on Neuronal Damage and Apoptotic Factors after Incomplete Cerebral Ischemia and Reperfusion in Rats: A Long-term Observation

Background: Propofol reduces neuronal damage from cerebral ischemia when investigated for less than 8 postischemic days. This study investigates the long-term effects of propofol on neuronal damage and apoptosis-related proteins after cerebral ischemia and reperfusion.

Methods: Male Sprague-Dawley rats were randomly assigned as follows: group 1 (n = 32, control): fentanyl and nitrous oxide-oxygen; group 2 (n = 32, propofol): propofol and oxygen-air. Ischemia (45 min) was induced by carotid artery occlusion and hemorrhagic hypotension. Pericranial temperature and arterial blood gases were maintained constant. After 1, 3, 7, and 28 postischemic days, brains were removed, frozen, and sliced. Hippocampal eosinophilic cells were counted. The amount of apoptosis-related proteins Bax, p53, Bcl-2, and Mdm-2 and neurons positive for activated caspase-3 were analyzed.

Results: In propofol-anesthetized rats, no eosinophilic neurons were detected, whereas in control animals, 16-54% of hippocampal neurons were eosinophilic (days 1-28). In control animals, the concentration of Bax was 70-200% higher after cerebral ischemia compared with that in animals receiving propofol over time. Bcl-2 was 50% lower in control animals compared with propofol-anesthetized rats during the first 3 days. In both groups, a maximal 3% of the hippocampal neurons were positive for activated caspase-3.     

Early detection of vascular dysfunction in type I diabetes

Blood flow and transcutaneous oxygen tension was measured by venous occlusion plethysmography and a transcutaneous oxygen electrode before and after 5 min of arterial occlusion in the forearm of young adult subjects with type I (insulin-dependent) diabetes without overt evidence of angiopathy. In control subjects (n = 21), the forearm blood flow increased by greater than or equal to 2.8-fold at 30 s after ischemia. Diabetic subjects with glycosylated hemoglobin (GHb) less than or equal to 9.5% (n = 15) exhibited a blood flow response that was not statistically different from normal control subjects. Diabetic subjects with GHb greater than or equal to 12.5% (n = 23) did not exhibit an increase in the postischemic blood flow. When blood flow patterns for the first 14 diabetic subjects were examined regardless of GHb value, four patterns of response were noted: 1) normal pattern (n = 3), 2) normal postischemic rise in blood flow with a prolonged elevation (n = 3), 3) no postischemic rise (n = 4), and 4) variable baseline blood flow with a decrease in blood flow postischemia (n = 4). This approach indicated that a comparison of means obscured potentially meaningful abnormal patterns. Abnormalities in the response of the transcutaneous oxygen tension to ischemia were observed in both groups of diabetic patients, but the difference between diabetic patients in good and poor control was less obvious. We have defined an abnormal response of blood flow and transcutaneous oxygen tension to ischemia that may correlate to glycemic control and have identified several patterns of blood flow after ischemia that may be important in defining the etiology and natural history of diabetic angiopathy.     

Therapeutic effects of hyperbaric oxygenation on acute focal cerebral ischemia in rats

The effects of hyperbaric oxygenation on acute focal cerebral ischemia in rats were investigated. All rats suffered 4-hour middle cerebral artery occlusion. Nontreated controls had a 27.9% +/- 5.5% infarct volume, and a left/right hemispheric volume ratio of 109% +/- 3%. Animals treated between 2.5 and 3.5 hours following occlusion had an 18.1% +/- 9.7% infarct volume (p less than 0.01), and a 105% +/- 3% left/right hemispheric volume ratio (p less than 0.001). In conclusion, at least until 4 hours following an ischemic insult, hyperbaric oxygenation reduces ischemic neuronal injury and brain edema following middle cerebral artery occlusion in rats treated between 2.5 and 3.5 hours following occlusion.     

Hyperglycemia exacerbates and insulin fails to protect in acute renal ischemia in the rat

Hyperglycemia worsens ischemic injury in several ischemic models. To determine whether renal lactate accumulation was associated with hyperglycemia-exacerbated postischemic renal dysfunction and mortality, halothane-anesthetized rats underwent right nephrectomy and 45 min of left renal artery and vein occlusion. Prior to ischemia, rats received saline (n = 22), glucose (2 g/kg, n = 22), or insulin (4 U/kg, n = 18). Sham-operated glucose-treated rats (2 g/kg, n = 4) underwent right nephrectomy and no vascular occlusion. As anticipated, glucose pretreatment elevated plasma glucose, while insulin pretreatment lowered plasma glucose; both were significantly different from values in saline controls. Creatinine was unchanged in sham-operated rats but was significantly higher in glucose-treated rats at 24 and 48 hr postischemia compared to saline controls. No statistical differences in creatinine were found when comparing saline controls to insulin-treated rats. Eighteen percent of glucose-treated rats survived to 72 hr postocclusion, while 45% of insulin-treated rats, 73% of saline control rats, and 100% of sham-operated rats survived this period. In a separate but identical treatment protocol, renal tissue was serially sampled and lactate content was determined in rats pretreated with saline (n = 7), glucose (n = 6) or insulin (n = 6) or sham-operated (n = 2) and receiving identical operation. Tissue lactate concentration did not change during serial sampling in the sham group. During ischemia, lactate was significantly higher in glucose-treated rats and significantly lower in insulin-treated rats as compared to saline controls. The adverse effects of exogenous glucose and attendant hyperglycemia on renal function during normothermic renal ischemia are demonstrated.(ABSTRACT TRUNCATED AT 250 WORDS)     

Dietary protein prior to renal ischemia dramatically affects postischemic kidney function

Male Sprague-Dawley rats were maintained on high protein (60%), normal protein (20%), low protein (5%), or no protein (0%) diets for two or four weeks prior to 45 minutes of renal ischemia induced by renal pedicle clamping. Most (93%) of the rats on the high protein diet died within three days following renal ischemia. In addition, 69% of the rats on normal protein diets also died, most before the fourth day following ischemic insult. In contrast, 88% of the rats on the low protein diet lived, although some exhibited elevated serum creatinine levels for up to one to two weeks following ischemia. Finally, all of the rats on no protein diets lived, and most (75%) exhibited normal serum creatinine levels by the fourth day following ischemia. Shifting the diets of high protein and normal protein adapted rats to no protein diets immediately following ischemia did not improve postischemic survival. Also, changing the diets of no protein adapted rats to high protein diets immediately following ischemia did not significantly affect postischemic recovery. When rats were maintained on no protein diets for shorter periods of time prior to ischemia, it was found that approximately a week on this diet is necessary to provide maximum protection from postischemic acute renal failure. These findings demonstrate a dramatic effect of dietary protein prior to ischemic induced acute renal failure, and suggests that preoperative dietary protein intake should be an important consideration in those situations which are predisposed to postoperative acute renal failure.