Oxidative Stress and Antioxidant Activity in Hypothermia and Rewarming. Can RONS Modulate the Beneficial Effects of Therapeutic Hypothermia?

Hypothermia is a condition in which core temperature drops below the level necessary to maintain bodily functions. The decrease in temperature may disrupt some physiological systems of the body, including alterations in microcirculation and reduction of oxygen supply to tissues. The lack of oxygen can induce the generation of reactive oxygen and nitrogen free radicals (RONS), followed by oxidative stress, and finally, apoptosis and/or necrosis. Furthermore, since the hypothermia is inevitably followed by a rewarming process, we should also consider its effects. Despite hypothermia and rewarming inducing injury, many benefits of hypothermia have been demonstrated when used to preserve brain, cardiac, hepatic, and intestinal function against ischemic injury. This review gives an overview of the effects of hypothermia and rewarming on the oxidant/antioxidant balance and provides hypothesis for the role of reactive oxygen species in therapeutic hypothermia

Extracellular ferritin contributes to neuronal injury in an in vitro model of ischemic stroke

Previous clinical and experimental studies have shown that neurological decline and poor functional outcome after acute ischemic stroke in humans are associated with high ferritin levels in serum and cerebrospinal fluid (CSF) within 24 hours of ischemic stroke onset. The aim of the present study was to find out if and how high extracellular ferritin concentrations can increase the excitotoxicity effect in a neuronal cortical culture model of stroke. Extracellular ferritin (100 ng/ml) significantly increased the excitotoxic effect caused by excessive exogenous glutamate (50 µM and 100 µM) by leading to an increase in lipid peroxidation, a reduction in mitochondrial membrane potential and a decrease in neuron viability. Extracellular apoferritin (100 ng/ml), the iron-free form of the protein, does not increase the excitotoxicity of glutamate, which proves that iron was responsible for the neurotoxic effect of the exogenous ferritin. We present evidence that extracellular ferritin iron exacerbate the neurotoxic effect induced by glutamate excitotoxicity and that the effect of ferritin iron is dependent of glutamate excitotoxicity. Our results support the idea that body iron overload is involved in the severity of the brain damage caused by stroke and reveal the need to control systemic iron homeostasis

Preconditioning-Like Properties of Short-Term Hypothermia in Isolated Perfused Rat Liver (IPRL) System

Hypothermia may attenuate the progression of ischemia-induced damage in liver. Here, we determined the effects of a brief cycle of hypothermic preconditioning applied before an ischemic/reperfusion (I/R) episode in isolated perfused rat liver (IPRL) on tissue damage and oxidative stress. Rats (male, 200-250 g) were anaesthetised with sodium pentobarbital (60 mg·kg-1 i.p) and underwent laparatomy. The liver was removed and perfused in a temperature-regulated non-recirculating system. Livers were randomly divided into two groups (n = 6 each group). In the hypothermia-preconditioned group, livers were perfused with hypothermic buffer (cycle of 10 min at 22 °C plus 10 min at 37 °C) and the other group was perfused at 37 °C. Both groups were then submitted to 40 min of warm ischemia and 20 min of warm reperfusion. The level of tissue-damage indicators (alanine amino transferase, ALT; lactate dehydrogenase, LDH; and proteins), oxidative stress markers (thiobarbituric acid-reactive substances, TBARS; advanced oxidation protein products, AOPP; and glutathione, GSH) were measured in aliquots of perfusate sampled at different time intervals. Histological determinations and oxidative stress biomarkers in homogenized liver (AOPP; TBARS; nitric oxide derivatives, NOx; GSH and glutathione disulphide, GSSG) were also made in the tissue at the end. Results showed that both damage and oxidant indicators significantly decreased while antioxidant increased in hypothermic preconditioned livers. In addition, homogenized liver determinations and histological observations at the end of the protocol corroborate the results in the perfusate, confirming the utility of the perfusate as a non-invasive method. In conclusion, hypothermic preconditioning attenuates oxidative damage and appears to be a promising strategy to protect the liver against IR injury

Ubiquitin-proteasome system and oxidative stress in liver transplantation

A major issue in organ transplantation is the development of a protocol that can preserve organs under optimal conditions. Damage to organs is commonly a consequence of flow deprivation and oxygen starvation following the restoration of blood flow and reoxygenation. This is known as ischemia-reperfusion injury (IRI): a complex multifactorial process that causes cell damage. While the oxygen deprivation due to ischemia depletes cell energy, subsequent tissue oxygenation due to reperfusion induces many cascades, from reactive oxygen species production to apoptosis initiation. Autophagy has also been identified in the pathogenesis of IRI, although such alterations and their subsequent functional significance are controversial. Moreover, proteasome activation may be a relevant pathophysiological mechanism. Different strategies have been adopted to limit IRI damage, including the supplementation of commercial preservation media with pharmacological agents or additives. In this review, we focus on novel strategies related to the ubiquitin proteasome system and oxidative stress inhibition, which have been used to minimize damage in liver transplantation

Subnormothermic perfusion in the isolated rat liver preserves the antioxidant glutathione and enhances the function of the ubiquitin proteasome system

The reduction of oxidative stress is suggested to be one of the main mechanisms to explain the benefits of subnormothermic perfusion against ischemic liver damage. In this study we investigated the early cellular mechanisms induced in isolated ratliversafter15minperfusionattemperaturesrangingfromnormothermia(37 ∘ C) to subnormothermia (26 ∘ Cand22 ∘ C). Subnormothermic perfusion was found to maintain hepatic viability. Perfusion at 22 ∘ C raised reduced glutathione levels and the activity of glutathione reductase; however, lipid and protein oxidation still occurred as determined by malondialdehyde, 4-hydroxynonenal-protein adducts, and advanced oxidation protein products. In livers perfused at 22 ∘ C the lysosomal and ubiquitin proteasome system (UPS) were both activated. The 26S chymotrypsin-like ( 훽 5) proteasome activity was significantly increased in the 26 ∘ C (46%) and 22 ∘ C (42%) groups. The increased proteasome activity may be due to increased Rpt6 Ser120 phosphorylation, which is known to enhance 26S proteasome activity. Together, our results indicate that the early events produced by subnormothermic perfusion in the liver can induce oxidative stress concomitantly with antioxidant glutathione preservation and enhanced function of the lysosomal and UPS systems. Thus, a brief hypothermia could trigger antioxidant mechanisms and may be functioning as a preconditioning stimulus

Intermittent Hypobaric Hypoxic Preconditioning Provides Neuroprotection by Increasing Antioxidant Activity, Erythropoietin Expression and Preventing Apoptosis and Astrogliosis in the Brain of Adult Rats Exposed to Acute Severe Hypoxia

Background: Exposure to intermittent hypoxia has been demonstrated to be an efficienttool for hypoxic preconditioning, preventing damage to cells and demonstrating therapeutic benefits.We aimed to evaluate the effects of respiratory intermittent hypobaric hypoxia (IHH) to avoid braininjury caused by exposure to acute severe hypoxia (ASH). Methods: biomarkers of oxidative damage,mitochondrial apoptosis, and transcriptional factors in response to hypoxia were assessed by Westernblot and immunohistochemistry in brain tissue. Four groups of rats were used: (1) normoxic (NOR),(2) exposed to ASH (FiO27% for 6 h), (3) exposed to IHH for 3 h per day over 8 days at 460 mmHg,and (4) ASH preconditioned after IHH. Results: ASH animals underwent increased oxidative-stress-related parameters, an upregulation in apoptotic proteins and had astrocytes with phenotype formscompatible with severe diffuse reactive astrogliosis. These effects were attenuated and even preventedwhen the animals were preconditioned with IHH. These changes paralleled the inhibition of NF-κBexpression and the increase of erythropoietin (EPO) levels in the brain. Conclusions: IHH exertedneuroprotection against ASH-induced oxidative injury by preventing oxidative stress and inhibitingthe apoptotic cascade, which was associated with NF-κB downregulation and EPO upregulatio

Cardiorespiratory, metabolic and muscular responses during a video-recorded aerobic dance session on an air dissipation platform

Background: Aerobic dance (AD) is an appropriate physical activity for improving cardiorespiratory fitness. This study aimed to compare cardiorespiratory and metabolic responses, and muscle fatigue between an air dissipation platform (ADP) and a hard surface during a video-recorded AD session. Methods: 25 healthy young women (23.3 ± 2.5 years) completed three sessions. In session 1, participants performed an incremental test to exhaustion on a treadmill. One week after session 1, participants were randomly assigned in a crossover design to perform video-recorded AD sessions on an ADP and on a hard surface (sessions 2 and 3). Cardiorespiratory and metabolic responses were assessed during AD sessions. Muscular fatigue was measured before and after AD sessions by a countermovement jump test. Results: Significantly higher heart rate, respiratory exchange ratio, pulmonary ventilation, ventilatory oxygen equivalent, and ventilatory carbon dioxide equivalent were observed on an ADP than on a hard surface (p 0.05). Conclusions: Video-recorded AD on an ADP increased the cardioventilatory and metabolic responses compared to a hard surface, preventing further muscle fatigue

Aerobic Dance on an Air Dissipation Platform Improves Cardiorespiratory, Muscular and Cellular Fitness in the Overweight and Obese Elderly

Background: Obesity is a global health problem associated with a high number of comorbidities that decrease functional capacity, especially in elderly people. Aerobic dance is considered a viable strategy to prevent the effects of aging, mainly in obese and overweight elderly people. This study aimed to evaluate the effects of aerobic dance on an air dissipation platform (ADP) on body composition, oxidative stress and muscular and cardiorespiratory fitness in elderly people. Methods: In total, 32 elderly adults (67.1 ± 3.6) were divided into 3 groups based on body mass index: healthy (HG), overweight (OWG) and obese (OG). Training program of aerobic dance on an ADP was carried out twice a week for 12 weeks. Results: OWG (p = 0.016) and OG decreased their weight (p < 0.001). There was a significant decrease in malondialdehyde concentrations in all experimental groups (p < 0.05). OWG and OG significantly improved their peak oxygen uptake (p < 0.01). HG increased the vertical jump height (p < 0.05), and HG and OG improved the power output of the lower extremities (p < 0.05). Conclusions: The aerobic dance on an ADP may be an effective alternative to lose weight, prevent oxidative stress and improve cardiorespiratory fitness in obese and overweight elderly people

The relevance of the UPS in the fatty liver graft preservation: a new approach for IGL-1 and HTK solutions

The 26S proteasome is the central proteolytic machinery of the ubiquitin proteasome system (UPS), which is involved in the degradation of ubiquitinated protein substrates. Recently, UPS inhibition has been shown to be a key factor in fatty liver graft preservation during organ cold storage using University of Wisconsin solution (UW) and Institute Georges Lopez (IGL-1) solutions. However, the merits of IGL-1 and histidine-tryptophan-ketoglutarate (HTK) solutions for fatty liver preservation have not been compared. Fatty liver grafts from obese Zücker rats were preserved for 24 h at 4 °C. Aspartate aminotransferase and alanine aminotransferase (AST/ALT), glutamate dehydrogenase (GLDH), ATP, adenosine monophosphate protein kinase (AMPK), e-NOS, proteasome activity and liver polyubiquitinated proteins were determined. IGL-1 solution prevented ATP breakdown during cold-storage preservation of steatotic livers to a greater extent than HTK solution. There were concomitant increases in AMPK activation, e-NOS (endothelial NOS (NO synthase)) expression and UPS inhibition. UPS activity is closely related to the composition of the solution used to preserve the organ. IGL-1 solution provided significantly better protection against ischemia-reperfusion for cold-stored fatty liver grafts than HTK solution. The effect is exerted through the activation of the protective AMPK signaling pathway, an increase in e-NOS expression and a dysregulation of the UPS

Aldehyde Dehydrogenase 2 (ALDH2) in Rat Fatty Liver Cold Ischemia Injury

Institut George Lopez-1 (IGL-1) and Histidine-tryptophan-ketoglutarate (HTK) solutions are proposed as alternatives to UW (gold standard) in liver preservation. Their composition differs in terms of the presence/absence of oncotic agents such as HES or PEG, and is decisive for graft conservation before transplantation. This is especially so when fatty (steatotic) livers are used since these grafts are more vulnerable to ischemia insult during conservation. Their composition determines the extent of the subsequent reperfusion injury after transplantation. Aldehyde dehydrogenase-2 (ALDH2), a mitochondrial enzyme, has been reported to play a protective role in warm ischemia-reperfusion injury (IRI), but its potential in fatty liver cold ischemic injury has not yet been investigated. We evaluated the relevance of ALDH2 activity in cold ischemia injury when fatty liver grafts from Zucker Obese rats were preserved in UW, HTK, and IGL-1 solutions, in order to study the mechanisms involved. ALDH2 upregulation was highest in livers preserved in IGL-1. It was accompanied by a decrease in transaminases, apoptosis (Caspase 3 and TUNEL assay), and lipoperoxidation, which was concomitant with the effective clearance of toxic aldehydes such as 4-hydroxy-nonenal. Variations in ATP levels were also determined. The results were consistent with levels of NF-E2 p45-related factor 2 (Nrf2), an antioxidant factor. Here we report for the first time the relevance of mitochondrial ALDH2 in fatty liver cold preservation and suggest that ALDH2 could be considered a potential therapeutic target or regulator in clinical transplantation