Loss Of Redox Homeostasis Research Articles

Sulodexide protects endothelial cells against 4-hydroxynonenal-induced oxidative stress and glutathione-dependent redox imbalance by modulation of sestrin2/nuclear factor erythroid 2-related factor 2 pathway.

The lipid peroxidation product 4-hydroxynonenal (HNE) may be involved in vascular endothelial cell damage by induction of oxidative stress, apoptosis, and loss of redox homeostasis. There is evidence that stimulation of endothelial cells with 4-HNE induces the activation of the nuclear factor erythroid 2-related factor 2/Kelch-like ECH-associated protein 1 (Nrf2/Keap-1) pathway. Sestrin2 protein (SESN2) is one of the key regulators of Nrf2 and is involved in the cellular response to oxidative stress. However, the function of SESN2 in HNE-induced endothelial injury is not yet understood. Sulodexide (SDX) is a mixture of glycosaminoglycans used in clinical practice in the treatment of chronic venous and arterial diseases. While SDX has well-documented endothelial protective properties, little is known about its antioxidant effects. The aim of this study was to elucidate the molecular mechanisms activated by SDX in human umbilical endothelial cells (HUVECs) under HNE-induced oxidative stress. In this experimental model, we decided to evaluate the anti-apoptotic and antioxidant potential of SDX and its effect on the SESN2/Nrf2/GSH pathway. HUVECs were treated with 25 _M HNE or HNE combined with 0.5 LRU/mL SDX for 4 hours. Cell viability, apoptosis and intracellular reactive oxygen species (ROS) production were assessed by MTT assay and fluorescence microscopy. The expressions of Bax, cleaved caspase-3, Keap-1 and Nrf2 were determined by Western blot analysis. The intracellular concentrations of reduced glutathione (GSH) and oxidized glutathione (GSSG) were measured by colorimetric assay. SESN2, glutamate-cysteine ligase catalytic subunit (GCLc) and glutathione synthase (GSS) were assessed using ELISA. RT-qPCR was performed to detect Nrf2, GCLc and GSS mRNA levels. Transient Nrf2 silencing was obtained by short interfering RNA (siRNA). We have demonstrated that SDX can reduce the negative impact of HNE on HUVECs. SDX significantly protected HNE-treated HUVECs from apoptosis (p<0.001) and oxidative stress (p<0.001). SDX treatment significantly reduced Bax (p<0.05) and cleaved caspase-3 (p<0.01) expression. Co-administration of HNE and SDX increased GSH content (p<0.001) and GSH:GSSG ratio (p<0.001) as well as decreased SESN2 concentration (p<0.001) and Nrf2 (p<0.01), GCLc (p<0.05) and GSS (p<0.01) gene expression compared with the HNE group. Moreover, we revealed a negative correlation between SESN2 levels and GSH concentrations (p<0.001). Nrf2 silencing significantly decreased the effect of HNE and SDX on the induction of GCLc and GSS genes. SDX also significantly ameliorated the increase of nuclear Nrf2 in response to HNE (p<0.05). The results confirmed that SDX may protect against HNE-induced endothelial damage through its antioxidant effect and modulation of the SESN2/Nrf2/GSH signaling pathway.

α-Ketoglutarate prevents hyperlipidemia-induced fatty liver mitochondrial dysfunction and oxidative stress by activating the AMPK-pgc-1α/Nrf2 pathway

α-Ketoglutarate (AKG), a crucial intermediate in the tricarboxylic acid cycle, has been demonstrated to mitigate hyperlipidemia-induced dyslipidemia and endothelial damage. While hyperlipidemia stands as a major trigger for non-alcoholic fatty liver disease, the protection of AKG on hyperlipidemia-induced hepatic metabolic disorders remains underexplored. This study aims to investigate the potential protective effects and mechanisms of AKG against hepatic lipid metabolic disorders caused by acute hyperlipidemia. Our observations indicate that AKG effectively alleviates hepatic lipid accumulation, mitochondrial dysfunction, and loss of redox homeostasis in P407-induced hyperlipidemia mice, as well as in palmitate-injured HepG2 cells and primary hepatocytes. Mechanistic insights reveal that the preventive effects are mediated by activating the AMPK-PGC-1α/Nrf2 pathway. In conclusion, our findings shed light on the role and mechanism of AKG in ameliorating abnormal lipid metabolic disorders in hyperlipidemia-induced fatty liver, suggesting that AKG, an endogenous mitochondrial nutrient, holds promising potential for addressing hyperlipidemia-induced fatty liver conditions.

Myokines: A central point in managing redox homeostasis and quality of life.

Redox homeostasis is a crucial phenomenon that is obligatory for maintaining the healthy status of cells. However, the loss of redox homeostasis may lead to numerous diseases that ultimately result in a compromised quality of life. Skeletal muscle is an endocrine organ that secretes hundreds of myokines. Myokines are peptides and cytokines produced and released by muscle fibers. Skeletal muscle secreted myokines act as a robust modulator for regulating cellular metabolism and redox homeostasis which play a prime role in managing and improving metabolic function in multiple organs. Further, the secretory myokines maintain redox homeostasis not only in muscles but also in other organs of the body via stabilizing oxidants and antioxidant levels. Myokines are also engaged in maintaining mitochondrial dynamics as mitochondria is a central point for the generation of reactive oxygen species (ROS). Ergo, myokines also act as a central player in communicating signals to other organs, including the pancreas, gut, liver, bone, adipose tissue, brain, and skin via their autocrine, paracrine, or endocrine effects. The present review provides a comprehensive overview of skeletal muscle-secreted myokines in managing redox homeostasis and quality of life. Additionally, probable strategies will be discussed that provide a solution for a better quality of life.

Combined effects of temperature rise and sodium lauryl sulfate in the Mediterranean mussel

Personal care products (PCPs) are those compounds used daily (e.g., soaps, shampoos, deodorants, and toothpaste), explaining their frequent detection in aquatic systems. Still, scarce information is available on their effects on inhabiting wildlife. Among the most commonly used PCPs is the surfactant Sodium Lauryl Sulfate (SLS). The present study investigated the influence of temperature (CTL 17 ºC vs 22 ºC) on the effects of SLS (0 mg/L vs 4 mg/L) in the mussel species Mytilus galloprovincialis. Mussels’ general health status was investigated, assessing their metabolic and oxidative stress responses. Higher biochemical alterations were observed in SLS-exposed mussels and warming enhanced the impacts, namely in terms of biotransformation capacity and loss of redox homeostasis, which may result in consequences to population maintenance, especially if under additional environmental stressors. These results confirm M. galloprovincialis as an excellent bioindicator of PCPs pollution, and the need to consider actual and predicted climate changes.

Modeling LMNA-associated Muscular Dystrophy in Drosophila and iPS Cell-Derived Cardiomyocytes Revealed Loss of Redox Homeostasis

Modeling LMNA-associated Muscular Dystrophy in Drosophila and iPS Cell-Derived Cardiomyocytes Revealed Loss of Redox Homeostasis

Food Processing, Dysbiosis, Gastrointestinal Inflammatory Diseases, and Antiangiogenic Functional Foods or Beverages.

Foods and beverages provide nutrients and alter the gut microbiota, resulting in eubiosis or dysbiosis. Chronic consumption of a diet that is high in saturated or trans fats, meat proteins, reducing sugars, and salt and low in fiber induces dysbiosis. Dysbiosis, loss of redox homeostasis, mast cells, hypoxia, angiogenesis, the kynurenine pathway, transglutaminase 2, and/or the Janus kinase pathway are implicated in the pathogenesis and development of inflammatory bowel disease, celiac disease, and gastrointestinal malignancy. This review discusses the effects of oxidative, carbonyl, or glycative stress-inducing dietary ingredients or food processing-derived compounds on gut microbiota and gastrointestinal epithelial and mast cells as well as on the development of associated angiogenic diseases, including key signaling pathways. The preventive or therapeutic potential and the biochemical pathways of antiangiogenic or proangiogenic foods or beverages are also described. The outcomes of the interactions between disease pathways and components of food are critical for the design of foods and beverages for healthy lives.

Can ocean warming alter sub-lethal effects of antiepileptic and antihistaminic pharmaceuticals in marine bivalves?

The negative effects induced in marine organisms by Climate Change related abiotic factors consequences, namely ocean warming, are well-known. However, few works studied the combined impacts of ocean warming and contaminants, as pharmaceutical drugs. Carbamazepine (CBZ) and cetirizine (CTZ) occur in the marine environment, showing negative effects in marine organisms. This study aimed to evaluate the impacts of ocean warming on the effects of CBZ and CTZ, when acting individually and combined (drug vs drug), in the edible clam Ruditapes philippinarum. For that, drugs concentration, bioconcentration factors and biochemical parameters, related with clam’s metabolic capacity and oxidative stress, were evaluated after 28 days exposure to environmentally relevant scenarios of these stressors. The results showed limited impacts of the drugs (single and combined) at control and warming condition. Indeed, it appeared that warming improved the oxidative status of contaminated clams (higher reduced to oxidized glutathione ratio, lower lipid peroxidation and protein carbonylation levels), especially when both drugs were combined. This may result from clam’s defence mechanisms activation and reduced metabolic capacity that, respectively, increased elimination and limited production of reactive oxygen species. At low stress levels, defence mechanisms were not activated which resulted into oxidative stress. The present findings highlighted that under higher stress levels clams may be able to activate defence strategies that were sufficient to avoid cellular damages and loss of redox homeostasis. Nevertheless, low concentrations were tested in the present study and the observed responses may greatly change under increased pollution levels or temperatures. Further research on this topic is needed since marine heat waves are increasing in frequency and intensity and pollution levels of some pharmaceuticals are also increasing in coastal systems.

How safe are the new green energy resources for marine wildlife? The case of lithium

Considering the increasing use of Lithium (Li) and the necessity to fulfil this demand, labile Li occurrence in the environment will be enhanced. Thus, additional research is needed regarding the presence of this element in marine environment and its potential toxic impacts towards inhabiting wildlife. The aim of the present study was to evaluate Li toxicity based on the exposure of Mytilus galloprovincialis to this metal, assessing the biochemical changes related with mussels’ metabolism, oxidative stress and neurotoxicity. For this, organisms were exposed to different Li concentrations (100, 250, 750 μg/L) for 28 days. The results obtained clearly demonstrated that Li lead to mussels’ metabolism depression. The present study also revealed that, especially at the highest concentrations, antioxidant and biotransformation enzymes were not activated, leading to the occurrence of lipid peroxidation and loss of redox homeostasis, with increased content in oxidized glutathione in comparison to the reduced form. Furthermore, after 28 days, higher Li exposure concentrations induced neurotoxic effects in mussels, with a decrease in acetylcholinesterase enzyme activity. The responses observed were closely related with Li concentrations in mussels’ tissues, which were more pronounced at higher exposure concentrations. Such results highlight the potential toxic effects of Li to marine species, which may even be higher under predicted climate changes and/or in the presence of other pollutants.

Redox-regulation of germination during imbibitional oxidative and chilling stress in an indica rice cultivar (Oryza sativa L., Cultivar Ratna).

Imbibitional oxidative stress of different magnitude, imposed by treatment with different titer of H2O2 (both elevated, 20mM and low, 500µM) to an indica rice cultivar (Oryza sativa L., Cultivar Ratna) caused formation of differential redox cues at the metabolic interface, as evident from significant alteration of ROS/antioxidant ratio, efficacy of ascorbate-glutathione cycle, radical scavenging property, modulation of total thiol content and expression of oxidative membrane protein and lipid damages as biomarkers of oxidative stress. All the redox parameters examined, substantiate the experimental outcome that treatment with elevated concentration of H2O2 caused serious loss of redox homeostasis and germination impairment, whereas low titre H2O2 treatment not only restored redox homeostasis but also improve germination and post-germinative growth. The inductive pulse of H2O2 (500µM) exhibited significantly better performance of ascorbate-glutathione pathway, which was otherwise down-regulated significantly in 20mM H2O2 treatment-raised seedlings. A comparison between imbibitional chilling stress-raised experimental rice seedlings with 20mM H2O2 treated rice seedling revealed similar kind of generation of redox cues and oxidative stress response. Further, imbibitional H2O2 treatments in rice also revealed a dose-dependent regulation of expression of genes of Halliwell-Asada pathway enzymes, which is in consonance with the redox metabolic response of germinating rice seeds. In conclusion, a dose-dependent regulation of H2O2 mediated redox cues and redox regulatory properties during germination in rice are suggested, the knowledge of which may be exploited as a promising seed priming technology.

Antioxidant Properties of Buffalo-Milk Dairy Products: A β-Lg Peptide Released after Gastrointestinal Digestion of Buffalo Ricotta Cheese Reduces Oxidative Stress in Intestinal Epithelial Cells.

Redox signaling regulates different gastrointestinal (G.I.) epithelium functions. At the intestinal level, the loss of redox homeostasis in intestinal epithelial cells (IECs) is responsible for the pathogenesis and development of a wide diversity of G.I. disorders. Thus, the manipulation of oxidative stress in IECs could represent an important pharmacological target for different diseases. In this study, peptides released from in vitro gastro intestinal digestion of different buffalo-milk commercial dairy products were identified and evaluated for their bioactive properties. In particular, six G.I. digests of dairy products were tested in a model of oxidative stress for IECs. Among them, buffalo ricotta cheese was the most active and the presence of an abundant β-lactoglobulin peptide (YVEELKPTPEGDL, f:60-72) was also revealed. The antioxidant potential of the identified peptide was also evaluated in a model of hydrogen peroxide (H2O2)-induced oxidative stress in the IEC-6 cell line. The peptide was able to reduce ROS release, while, on the other hand, it increased nuclear factor (erythroid-derived 2)-like 2 (Nrf2) activation and the expression of antioxidant cytoprotective factors, such as heme oxygenase 1 (HO-1), NAD(P)H:quinone oxidoreductase 1 (NQO1), and superoxide dismutase (SOD). These results indicate that buffalo ricotta cheese-isolated peptide could have potential in the treatment of some gastrointestinal disorders.

CARDIOVASCULAR RISK STRATIFICATION IN YOUNG POPULATION

Objective: Cardiovascular (CV) diseases are the leading cause of death in developed countries and present high prevalence in developing countries. Nevertheless, there are no biomarkers available to detect the causal physiopathological process of the clinical illness, implying a high socioeconomic impact. Nowadays, CV risk estimators focus on age as the most important risk factor in CV events and mortality. The current study is focused on CV risk stratification attending to oxidative stress for the discovery of new markers for early detection of the physiopathological process. Design and method: 24 blood samples from a cohort of patients in average age of life (30–50 years old) classified in 3 groups of study according to their CV risk (healthy controls, patients with CV risk factors and patients with reported CV event in the last 3 years) were analyzed using FASILOX, a novel strategy for analysis of the dynamic thiol redox proteome. To our knowledge, this study constitutes the first description of reversible Cys oxidation in human plasma according to CV risk stratification. Results: We identified that reversible oxidations are decreased in CV events individuals compared with healthy subjects. Only proteins involved in coagulation cascade have reversible oxidations increased in CV events, biologic function directly linked with CV pathology, whose alteration could result in a CV event.We have also showed down-regulation of proteins involved in antioxidant respond, which is in line with the loss of redox homeostasis in CV events. Conclusions: CV events present a severe oxidative stress, exceeding antioxidant mechanisms as reversible oxidations of Cys, generating irreversible oxidations that together with the down-regulation of redoxins, generate an increase of molecular damage.The present draft of redox targets together with the quantification of protein and oxidative changes may help to better

Uterine protein oxidative modifications may condition trophoblast function

Aim: Evaluate whether protein carbonylation resulting from uterine altered redox imbalance interferes with extravillous trophoblast viability. Introduction: Local redox homeostasis is believed to have a pivot role in uterine transformation necessary for blastocyst implantation and placenta development. By contrast, redox status imbalance plays a role in deficient placentation and the development of pregnancy-related complications (e.g. preeclampsia or gestational diabetes) with increased incidence in older women. Thus, it was hypothesized that at an older reproductive age, loss of redox homeostasis is a contributor to disruption of foetal/placental interactions and the development of such complications. Methods: Uterine human samples were collected at delivery by elective caesarean section. The protocol was approved by the ethical committee of “Centro Materno-Infantil do Porto”, volunteers gave written consent to be included in the study. Total protein carbonylation was detected by oxyblot and protein expression was quantified by western blotting. Specific protein carbonylation was verified by immunoprecipitation. Albumin was carbonylated using hydrogen peroxide (H2O2), followed by dialysis, and western blotting to confirm carbonylated albumin. HST-8SV neo extravillous trophoblasts were treated with carbonylated/non-carbonylated albumin, followed by cell viability assay. A P value less than 0.05 was assumed to denote significant difference. Results: At the placental site, carbonylated albumin normalized to total albumin expression showed a positive and significant association with maternal age. (r=0.6909, P=0.0021) In vitro, carbonylated albumin displayed a cytotoxic effect, at concentrations ranging from 10 to 100μg/ml. Lower concentrations did not affect trophoblast viability. Conclusion: Uterine aging is accompanied by selective albumin oxidative modifications, which appears to interfere with trophoblast ability to invade and transform the maternal placental site.

Chemistry‐Driven Approaches to Deconstruct Redox‐Linked Signal Transduction Networks

Oxidative and electrophilic stress that results from loss of redox homeostasis has become increasingly appreciated to underlie numerous disease states including inflammation, cardiovascular disease, cancer, neurodegeneration, and psychiatric disorders. Utilizing our novel T‐REX (Targeted Reactive Electrophiles and oXidants) methodology, we aim to better resolve electrophile‐ and oxidant‐mediated signaling effects on any protein of interest (POI). In contrast to widely used methodologies, in which live specimens are treated globally with inherently reactive electrophiles and oxidants (RES/ROS) in excess quantities, our technique allows us to selectively deliver these reactive species to specific POIs with spatiotemporal control. Herein, T‐REX is exploited to delineate the functional consequences of H2O2 signaling along individual disease relevant redox‐dependent pathways in live cells and animals.

IDH2 deficiency promotes mitochondrial dysfunction and cardiac hypertrophy in mice

Cardiac hypertrophy, a risk factor for heart failure, is associated with enhanced oxidative stress in the mitochondria, resulting from high levels of reactive oxygen species (ROS). The balance between ROS generation and ROS detoxification dictates ROS levels. As such, disruption of these processes results in either increased or decreased levels of ROS. In previous publications, we have demonstrated that one of the primary functions of mitochondrial NADP+-dependent isocitrate dehydrogenase (IDH2) is to control the mitochondrial redox balance, and thereby mediate the cellular defense against oxidative damage, via the production of NADPH. To explore the association between IDH2 expression and cardiac function, we measured myocardial hypertrophy, apoptosis, and contractile dysfunction in IDH2 knockout (idh2−/−) and wild-type (idh2+/+) mice. As expected, mitochondria from the hearts of knockout mice lacked IDH2 activity and the hearts of IDH2-deficient mice developed accelerated heart failure, increased levels of apoptosis and hypertrophy, and exhibited mitochondrial dysfunction, which was associated with a loss of redox homeostasis. Our results suggest that IDH2 plays an important role in maintaining both baseline mitochondrial function and cardiac contractile function following pressure-overload hypertrophy, by preventing oxidative stress.

Oxidative stress in the cardiorenal metabolic syndrome.

Excess visceral adiposity contributes to inappropriate activation of the renin-angiotensin-aldosterone system despite a state of volume expansion and of salt retention that contributes to subclinical elevations of pro-oxidant mechanisms. These adverse effects are mediated by excess generation of reactive oxygen species (ROS) and diminished antioxidant defense mechanisms. Excess tissue (i.e., skeletal muscle, liver, heart) free oxygen radicals contribute to impairments in the insulin-dependent metabolic signaling pathways that regulate glucose utilization/disposal and systemic insulin sensitivity. The generation of ROS is required for normal cell signaling and physiological responses. It is a loss of redox homeostasis that results in a proinflammatory/profibrotic milieu that promotes impairments in insulin metabolic signaling, reduced endothelial-mediated vasorelaxation, and associated cardiovascular and renal structural and functional abnormalities. These maladaptive processes are increasingly recognized as important in the progression of hypertension in the cardiorenal metabolic phenotype. There is increasing evidence to support a critical role for Ang II signaling through the AT(1)R and aldosterone actions through the MR in conjunction with an altered redox-mediating impaired endothelial, cardiac and renal function in this metabolic phenotype. There are emerging clinical data that indicate that therapies that target the renin angiotensin-aldosterone system (RAAS) also attenuate oxidative stress, and improve endothelial, cardiac and renal functions, which collectively contribute to reductions in hypertension.