Lower doses of isoflurane treatment has no beneficial effects in a rat model of intracerebral hemorrhage

Background: Intracerebral hemorrhage is a subtype of stroke that has a poor prognosis without an adequate therapy. Recently, the use of anesthetics such as isoflurane has been shown to be protective after cerebral ischemia. However, the potential therapeutic effect of isoflurane after intracerebral hemorrhage (ICH) has not been fully explored. Results: In this study, male Sprague–Dawley rats (SD) were subjected to ICH and randomized into controls and 1.2% or 1.5% isoflurane posttreatment groups. Brain water content, neurological outcomes and matrix metalloproteinase-2 and -9 (MMP2-MMP9) plasma levels were quantified at 24 hours. Isoflurane treatment did not reduce brain edema compared with controls in any of the applied isoflurane concentrations. Moreover, consistent with this lack of effect on brain edema, isoflurane posttreatment did not affect neurological outcomes in any of the tests used. Plasma MMP levels did not change. Conclusion: Our data suggested that there is no neuroprotection after isoflurane posttreatment in a rat model of ICH

Diurnal Differences in Immune Response in Brain, Blood and Spleen After Focal Cerebral Ischemia in Mice.

BACKGROUND The immune response to acute cerebral ischemia is a major factor in stroke pathobiology. Circadian biology modulates some aspects of immune response. The goal of this study is to compare key parameters of immune response during the active/awake phase versus inactive/sleep phase in a mouse model of transient focal cerebral ischemia. METHODS Mice were housed in normal or reversed light cycle rooms for 3 weeks, and then they were blindly subjected to transient focal cerebral ischemia. Flow cytometry was used to examine immune responses in blood, spleen, and brain at 3 days after ischemic onset. RESULTS In blood, there were higher levels of circulating T cells in mice subjected to focal ischemia during zeitgeber time (ZT)1-3 (inactive or sleep phase) versus ZT13-15 mice (active or awake phase). In the spleen, organ weight and immune cell numbers were lower in ZT1-3 versus ZT13-15 mice. Consistent with these results, there was an increased infiltration of activated T cells into brain at ZT1-3 compared with ZT13-15. CONCLUSIONS This proof-of-concept study indicates that there are significant diurnal effects on the immune response after focal cerebral ischemia in mice. Hence, therapeutic strategies focused on immune targets should be reassessed to account for the effects of diurnal rhythms and circadian biology in nocturnal rodent models of stroke.Supported in part by the Rappaport Foundation and Leducq Foundation. The authors thank all team members of the MGH animal facility for help with light schedule switching.S

Anesthesia and Surgery Impair Blood–Brain Barrier and Cognitive Function in Mice

Blood–brain barrier (BBB) dysfunction, e.g., increase in BBB permeability, has been reported to contribute to cognitive impairment. However, the effects of anesthesia and surgery on BBB permeability, the underlying mechanisms, and associated cognitive function remain largely to be determined. Here, we assessed the effects of surgery (laparotomy) under 1.4% isoflurane anesthesia (anesthesia/surgery) for 2 h on BBB permeability, levels of junction proteins and cognitive function in both 9- and 18-month-old wild-type mice and 9-month-old interleukin (IL)-6 knockout mice. BBB permeability was determined by dextran tracer (immunohistochemistry imaging and spectrophotometric quantification), and protein levels were measured by Western blot and cognitive function was assessed by using both Morris water maze and Barnes maze. We found that the anesthesia/surgery increased mouse BBB permeability to 10-kDa dextran, but not to 70-kDa dextran, in an IL-6-dependent and age-associated manner. In addition, the anesthesia/surgery induced an age-associated increase in blood IL-6 level. Cognitive impairment was detected in 18-month-old, but not 9-month-old, mice after the anesthesia/surgery. Finally, the anesthesia/surgery decreased the levels of β-catenin and tight junction protein claudin, occludin and ZO-1, but not adherent junction protein VE-cadherin, E-cadherin, and p120-catenin. These data demonstrate that we have established a system to study the effects of perioperative factors, including anesthesia and surgery, on BBB and cognitive function. The results suggest that the anesthesia/surgery might induce an age-associated BBB dysfunction and cognitive impairment in mice. These findings would promote mechanistic studies of postoperative cognitive impairment, including postoperative delirium

Transient limb ischemia induces remote postconditioning and protects the brain from stroke induced damage

ABSTRACT Background. It has been recently hypothesized that a sub-lethal ischemic insult induced in an organ is able to protect from a harmful ischemia occurring in a different and anatomically distant organ. In this paper, a new neuroprotective strategy termed remote ischemic postconditioning is described and characterized for the first time. This neuroprotective mechanism occurs in rats in which a harmful brain ischemia is followed by a sub-lethal ischemic insult applied at the femoral artery level. Methods and Results. Remote ischemic postconditioning was induced in adult male rats by subjecting them to 100 minutes of middle cerebral artery occlusion (MCAO) followed by several brief cycles of ipsilateral femoral artery occlusion-reperfusion. Within all the considered experimental protocols, the one in which 100 minutes of MCAO were followed by 10 minutes reperfusion and 20 minutes occlusion of the femoral artery was the most effective in reducing brain infarct, resulting in almost 50% reduction in the infarct volume if compared to animals subjected to 100’ MCAO alone. Importantly, this protection was still present 7 days after remote postconditioning induction. Experiments carried out with specific inhibitors of each NO synthesizing enzyme, demonstrated that NO production through nNOS mediates part of the neuroprotective effect induced by remote ischemic postconditioning. In fact, the pharmacological blockade of nNOS was able to partially revert the neuroprotection induced by remote postconditioning. In addition, the neuroprotection induced by remote postconditioning was partially reverted when ganglion transmission was pharmacologically interrupted by hxamethonium, thus showing that neural factors are involved in this neuroprotective process. Conclusion. Collectively, the results of the present study demonstrate that remote postconditioning induces a marked neuroprotection through nNOS activation and it may represent a new clinically feasible therapeutic approach to treat ischemic stroke

In cold blood: intraarteral cold infusions for selective brain cooling in stroke

Hypothermia is a promising therapeutic option for stroke patients and an established neuroprotective treatment for global cerebral ischemia after cardiac arrest. While whole body cooling is a feasible approach in intubated and sedated patients, its application in awake stroke patients is limited by severe side effects: Strong shivering rewarms the body and potentially worsens ischemic conditions because of increased O(2) consumption. Drugs used for shivering control frequently cause sedation that increases the risk of aspiration and pneumonia. Selective brain cooling by intraarterial cold infusions (IACIs) has been proposed as an alternative strategy for patients suffering from acute ischemic stroke. Preclinical studies and early clinical experience indicate that IACI induce a highly selective brain temperature decrease within minutes and reach targeted hypothermia 10 to 30 times faster than conventional cooling methods. At the same time, body core temperature remains largely unaffected, thus systemic side effects are potentially diminished. This review critically discusses the limitations and side effects of current cooling techniques for neuroprotection from ischemic brain damage and summarizes the available evidence regarding advantages and potential risks of IACI

Pharmacologic pre- and postconditioning for stroke: Basic mechanisms and translational opportunity

Beyond reperfusion therapies, there are still no widely effective therapies for ischemic stroke. Although much progress has been made to define the molecular pathways involved, targeted neuroprotective strategies have often failed in clinical trials. An emerging hypothesis suggests that focusing on single targets and mechanisms may not work since ischemic stroke triggers multiple pathways in multiple cell types. In this review, we briefly survey and assess the opportunities that may be afforded by pre- and postconditioning therapies, with particular attention to pharmacologic pre- and postconditioning. Pharmacologic conditioning may be defined as the use of chemical agents either before or shortly after stroke onset to trigger mechanisms of endogenous tolerance that are thought to involve evolutionarily conserved signals that offer broad protection against ischemia. Importantly, many of the pharmacologic agents may also have been previously used in humans, thus providing hope for translating basic mechanisms into clinical applications

ASIC1a contributes to neuroprotection elicited by ischemic preconditioning and postconditioning

Acid-sensing ion channels, ASICs, are proton-gated cation channels widely expressed in peripheral sensory neurons and in neurons of the central nervous system that play an important role in a variety of physiological and pathological processes. To further confirm the role played by ASIC1a in cerebral ischemia, here we examined the involvement of this channel in two endogenous recently characterized neuroprotective strategies: brain ischemic preconditioning and postconditioning. The main aim of this study was to elucidate whether ASIC1a might take part as effector in the neuroprotection evoked by brain ischemic preconditioning and postconditioning. For this purpose we investigated the effect of ischemic preconditioning and postconditioning on (1) ASIC1a mRNA and protein expression in the temporoparietal cortex of rats at different time intervals; and (2) the effect of p-AKT inhibition on ASIC1a expression during ischemic preconditioning and postconditioning. Ischemic preconditioning and postconditioning were experimentally induced in adult male rats by subjecting them to different protocols of middle cerebral artery occlusion and reperfusion. ASIC1a expression was dramatically reduced in both the neuroprotective processes. These changes in ASIC expression were p-AKT mediated, since LY-294002, a specific p-AKT inhibitor, was able to prevent variations in ASIC1a expression. The results of the present study support the idea that the downregulation of ASIC1a expression and activity might be a reasonable strategy to reduce the infarct extension after stroke

Glymphatic and lymphatic communication with systemic responses during physiological and pathological conditions in the central nervous system

Abstract Crosstalk between central nervous system (CNS) and systemic responses is important in many pathological conditions, including stroke, neurodegeneration, schizophrenia, epilepsy, etc. Accumulating evidence suggest that signals for central-systemic crosstalk may utilize glymphatic and lymphatic pathways. The glymphatic system is functionally connected to the meningeal lymphatic system, and together these pathways may be involved in the distribution of soluble proteins and clearance of metabolites and waste products from the CNS. Lymphatic vessels in the dura and meninges transport cerebrospinal fluid, in part collected from the glymphatic system, to the cervical lymph nodes, where solutes coming from the brain (i.e., VEGFC, oligomeric α-syn, β-amyloid) might activate a systemic inflammatory response. There is also an element of time since the immune system is strongly regulated by circadian rhythms, and both glymphatic and lymphatic dynamics have been shown to change during the day and night. Understanding the mechanisms regulating the brain-cervical lymph node (CLN) signaling and how it might be affected by diurnal or circadian rhythms is fundamental to find specific targets and timing for therapeutic interventions