Main Non-enzymatic Antioxidant Research Articles

Mechanisms and regulations of ferroptosis.

Regulation of cell mortality for disease treatment has been the focus of research. Ferroptosis is an iron-dependent regulated cell death whose mechanism has been extensively studied since its discovery. A large number of studies have shown that regulation of ferroptosis brings new strategies for the treatment of various benign and malignant diseases. Iron excess and lipid peroxidation are its primary metabolic features. Therefore, genes involved in iron metabolism and lipid metabolism can regulate iron overload and lipid peroxidation through direct or indirect pathways, thereby regulating ferroptosis. In addition, glutathione (GSH) is the body's primary non-enzymatic antioxidants and plays a pivotal role in the struggle against lipid peroxidation. GSH functions as an auxiliary substance for glutathione peroxidase 4 (GPX4) to convert toxic lipid peroxides to their corresponding alcohols. Here, we reviewed the researches on the mechanism of ferroptosis in recent years, and comprehensively analyzed the mechanism and regulatory process of ferroptosis from iron metabolism and lipid metabolism, and then described in detail the metabolism of GPX4 and the main non-enzymatic antioxidant GSH in vivo.

Nanoscale coordination polymers enabling antioxidants inhibition for enhanced chemodynamic therapy.

Reactive oxygen species (ROS) generation to induce cell death is an effective strategy for cancer therapy. In particular, chemodynamic therapy (CDT), using Fenton-type reactions to generate highly cytotoxic hydroxyl radical (•OH), is a promising treatment modality. However, the therapeutic efficacy of ROS-based cancer treatment is still limited by some critical challenges, such as overexpression of enzymatic and non-enzymatic antioxidants by tumor cells, as well as the low tumor targeting efficiency of therapeutic agents. To address those problems, biomimetic CuZn protoporphyrin IX nanoscale coordination polymers have been developed, which significantly amplify oxidative stress against tumors by simultaneously inhibiting enzymatic and non-enzymatic antioxidants and initiating the CDT. In this design, cancer cell membrane camouflaged nanoparticle exhibits an excellent homotypic targeting effect. After being endocytosed into tumor cells, the nanoparticles induce depletion of the main non-enzymatic antioxidant glutathione (GSH) by undergoing a redox reaction with GSH. Afterward, the redox reaction generated cuprous ion (Cu+) works as a CDT agent for •OH generation. Furthermore, the released Zn protoporphyrin IX strongly inhibits the activity of the typical enzymatic antioxidant heme oxygenase-1. This tetra-modal synergistic strategy endows the biomimetic nanoparticles with great capability for anticancer therapy, which has been demonstrated in both in vitro and in vivo studies.

Physiological Responses of a Diazotrophic Cyanobacterium to Acidification of Paddy Floodwater: N2 Fixation, Photosynthesis, and Oxidative-Antioxidative Characteristics.

Long-term of excessive fertilization using nitrogen (N) chemical fertilizer caused the acidification of paddy soils. Presently, the impacts of soil acidification on physiological characteristics of diazotrophic cyanobacteria remain unknown. In order to elucidate this issue, the effects of paddy floodwater acidification on activities of respiration, photosynthetic oxygen evolution, and N2 fixation of a paddy diazotrophic cyanobacterium Aliinostoc sp. YYLX235 were investigated in this study. In addition, the origination and quenching of intracellular reactive oxygen species (ROS) were analyzed. The acidification of paddy floodwater decreased intracellular pH and interfered in energy flux from light-harvesting chlorophyll antenna to the reaction center of photosystem II (PS II). Activities of respiration, photosynthetic oxygen evolution, and N2 fixation were decreased by the acidification of paddy floodwater. Accompanied with an increase in ROS, the level of antioxidative system increased. Superoxide dismutase (SOD) and catalase (CAT) were the main enzymatic ROS scavengers in the cells of YYLX235; reduced glutathione (GSH) was the main non-enzymatic antioxidant. Antioxidants and oxidants in the cells of YYLX235 lost balance when the pH of paddy floodwater fell to 5.0 and 4.0, and lipid oxidative damage happened. The results presented in this study suggest that the acidification of paddy soil severely interfered in the photosynthesis of diazotrophic cyanobacteria and induced the production of ROS, which in turn resulted in oxidative damage on diazotrophic cyanobacteria and a decrease in cell vitality.

Computational Analysis of Alternative Photosynthetic Electron Flows Linked With Oxidative Stress.

During photosynthesis, organisms respond to their energy demand and ensure the supply of energy and redox equivalents that sustain metabolism. Hence, the photosynthetic apparatus can, and in fact should, be treated as an integrated supply-demand system. Any imbalance in the energy produced and consumed can lead to adverse reactions, such as the production of reactive oxygen species (ROS). Reaction centres of both photosystems are known sites of ROS production. Here, we investigate in particular the central role of Photosystem I (PSI) in this tightly regulated system. Using a computational approach we have expanded a previously published mechanistic model of C3 photosynthesis by including ROS producing and scavenging reactions around PSI. These include two water to water reactions mediated by Plastid terminal oxidase (PTOX) and Mehler and the ascorbate-glutathione (ASC-GSH) cycle, as a main non-enzymatic antioxidant. We have used this model to predict flux distributions through alternative electron pathways under various environmental stress conditions by systematically varying light intensity and enzymatic activity of key reactions. In particular, we studied the link between ROS formation and activation of pathways around PSI as potential scavenging mechanisms. This work shines light on the role of alternative electron pathways in photosynthetic acclimation and investigates the effect of environmental perturbations on PSI activity in the context of metabolic productivity.

Evaluation of Sub-Lethal Toxicity of Benzethonium Chloride in Cyprinus carpio Liver

Benzenthonium chloride (BEC, Hyamine 1622) is a quaternary ammonium surfactant with cationic properties widely used in cleaning, sanitation, and medical products that can become harmful to humans and also to the environment. This study aimed to evaluate its acute effects on Cyprinus carpio fish in terms of oxidative stress and morphological changes on hepatic tissue in order to show the sub-lethal toxicity of BEC. Fish were exposed to 1 mg/L BEC for 24, 48, and 96 h, and the liver samples were collected. The most significant changes were noticed after 96 h of exposure when the entire antioxidant enzyme system was affected. The activities of catalase, glutathione peroxidase, glutathione reductase, and glutathione S-transferase decreased by 44%, 31%, 30%, and 45%, respectively, compared to control. Glucose-6-phosphate dehydrogenase activity decreased by 29% after 96 h of control, inducing a reduction of NADPH formation which decreased by half the level of reduced glutathione, the main non-enzymatic antioxidant. These effects correlated with the raised value of lipid peroxidation after 96 h and the morphology changes on hepatic tissue, such as cytoplasmic vacuolization and nuclear hypertrophy that could affect the normal function of the liver. All of these results showed acute toxicity of BEC on C. carpio after 96 h of exposure, causing oxidative stress response at the hepatic level.

Glutathione and Nitric Oxide: Key Team Players in Use and Disuse of Skeletal Muscle.

Glutathione (GSH) is the main non-enzymatic antioxidant playing an important role in detoxification, signal transduction by modulation of protein thiols redox status and direct scavenging of radicals. The latter function is not only performed against reactive oxygen species (ROS) but GSH also has a fundamental role in buffering nitric oxide (NO), a physiologically-produced molecule having-multifaceted functions. The efficient rate of GSH synthesis and high levels of GSH-dependent enzymes are characteristic features of healthy skeletal muscle where, besides the canonical functions, it is also involved in muscle contraction regulation. Moreover, NO production in skeletal muscle is a direct consequence of contractile activity and influences several metabolic myocyte pathways under both physiological and pathological conditions. In this review, we will consider the homeostasis and intersection of GSH with NO and then we will restrict the discussion on their role in processes related to skeletal muscle function and degeneration.

Potential therapeutic role of melatonin in traumatic brain injury: A literature review

Oxidative stress induces excessive production of reactive oxygen species (ROS). ROS are including several free oxygen radicals such as singlet oxygen and superoxide radical. Excessive ROS production induces injuries of lipids, nucleic acids and proteins in several cells. Brain and neurons have a high amount of polyunsaturated fatty acids (PUFAs) and consumption of oxygen, but they have low level of antioxidant. Oxidative stress is controlled by several enzymatic and non-enzymatic antioxidants. One of the main nonenzymatic antioxidant is melatonin. Melatonin is secreted from the pineal gland by physiological circadian cycles. It has several physiological functions such as mediator of circannual reproductive rhythms (Tamtaji et al. 2019). However, it has also a regulatory role in the pathophysiological pathways of traumatic brain injury (TBI) in human and rodents (Barlow et al. 2019). TBI is one of the most common causes of the mortalities. Secondary events occur after primary events like shearing of nerve cells and blood vessels, cause posttraumatic neurodegenerations with an increase in ROS and ROS-mediated lipid peroxidation. It was reported that TBI-induced oxidative stress in experimental TBI was inhibited by the melatonin treatment (Senol and Nazıroğlu, 2014). Results of a recent study indicated protective role of melatonin through inhibition of Nrf2 signaling pathway, inflammation and oxidative stress in TBI-induced mice (Wang et al. 2019). In human studies, behavioral outcomes of TBI were modulated by the melatonin treatment (Barlow et al. 2019). In the oral presentation, I will review recent studies on TBI in human and experimental animals.   In conclusion, there are pre-clinical and clinical evidences that melatonin treatment after TBI significantly improves both behavior-cognition outcomes and pathophysiological outcomes such as oxidative stress and inflammation. It seems that the certain interaction between melatonin and TBI still remain to be determined.

Blood oxidative stress and metallothionein expression in Rett syndrome: Probing for markers

Objectives: Oxidative stress seems to be involved in Rett syndrome (RTT). The aim of this study was to assess the antioxidant status in RTT children with MECP2 gene mutations with respect to healthy controls, and to explore novel blood antioxidant markers for RTT severity. Methods: In erythrocytes from RTT females aged 2–14 years (n = 27) and age-matched controls (n = 27), we measured the levels of malonaldehyde and the activity of two antioxidant enzymes, Cu/Zn-superoxide dismutase and catalase, by spectrophotometric assays. In leukocytes, the expression of metallothioneins, the main non-enzymatic antioxidants, was assessed by real-time RT-PCR. In nine selected RTT children, methylome analysis was also performed. Results: Blood of RTT patients showed increased lipid peroxidation and a dysregulated pattern of MT expression, while enzymatic activities did not change significantly with respect to controls. Moreover, we observed no epigenetic dysregulation in CpG-enriched promoter regions of the analysed genes but significant hypomethylation in the random loci. Conclusions: As the haematic level of MT-1A directly correlates with the phenotype severity, this metallothionein can represent a marker for RTT severity. Moreover, the attempt to link the level of blood oxidative stress with MECP2 mutation and specific clinical features led us to draw some interesting conclusions.

Protective Effect of Quercetin on the Development of Preimplantation Mouse Embryos against Hydrogen Peroxide-Induced Oxidative Injury

Quercetin, a plant-derived flavonoid in Chinese herbs, fruits and wine, displays antioxidant properties in many pathological processes associated with oxidative stress. However, the effect of quercetin on the development of preimplantation embryos under oxidative stress is unclear. The present study sought to determine the protective effect and underlying mechanism of action of quercetin against hydrogen peroxide (H2O2)-induced oxidative injury in mouse zygotes. H2O2 treatment impaired the development of mouse zygotes in vitro, decreasing the rates of blastocyst formation and hatched, and increasing the fragmentation, apoptosis and retardation in blastocysts. Quercetin strongly protected zygotes from H2O2-induced oxidative injury by decreasing the reactive oxygen species level, maintaining mitochondrial function and modulating total antioxidant capability, the activity of the enzymatic antioxidants, including glutathione peroxidase and catalase activity to keep the cellular redox environment. Additionally, quercetin had no effect on the level of glutathione, the main non-enzymatic antioxidant in embryos.

Effect of short-term cryostimulation on antioxidative status and its clinical applications in humans.

Whole body cryostimulation (WBCT) is becoming popular in medicine and sport as an adjuvant form of treatment since late 1970s. Only a few works concerning antioxidant protection after WBCT have been published. The aim of this study was to determine the effect of a ten 3-min-long exposures (one exposure per day) to cryogenic temperature (−130°C) on the level of total antioxidant status (TAS), activity of selected antioxidant enzyme superoxide dismutase (SOD) and main non-enzymatic antioxidant—uric acid (UA) in WBCT study group (man n = 24; female n = 22) and non-WBCT control subjects (man n = 22; female n = 26). Moreover, we evaluated the lipid peroxidation measured as thiobarbituric acid reactive substances products. Their blood samples were collected twice at an interval of 10 days in both study group and control subjects. The activity of antioxidant enzyme and lipid peroxidation was assayed in erythrocytes, while the concentration of uric acid was measured in plasma. After completing a total of ten WBCT sessions a significant increase (p < 0.001) of TAS and UA levels in plasma (p < 0.001) in comparison to non-WBCT was observed. Our data showed that there was statistically significant increase of the activities of SOD in erythrocytes obtained from WBCT study group compared to non-WBCT controls after 10 days of treatment (p < 0.001). It was concluded that expositions to extremely low temperatures use in cryostimulation improve the antioxidant capacity of organism.

Oxylipin profile and antioxidant status of potato tubers during extended storage at room temperature

Potato tubers (cv. Bintje) (Solanum tuberosum L.) were stored under extreme conditions at 20 degrees C for 350 days without sprout inhibitors in order to assess whether aging- and/or senescence-related processes occurred. Under these extreme storage conditions, multiple sprouting followed by the formation of daughter tubers occurs. At the same time, an increase in respiration intensity, as evidenced by cytochrome c oxidase activity (E.C. 1.9.3.1), is observed, leading to a potential increase in reactive oxygen species (ROS) production. As polyunsaturated fatty acids are priority targets of oxidative attacks, the damage to lipids was assessed by oxylipin profiling in both free and esterified forms. Oxylipin profiling showed a predominance of linoleic acid-derived oxylipins and of 9-hydroxy and 9-hydroperoxy fatty acids in both free and esterified forms. No significant accumulation of individual oxylipin was observed 350 days after harvest. To further understand the absence of lipid breakdown products accumulation, the main enzymatic and non-enzymatic antioxidants were assessed. Antioxidant enzyme activities [superoxide dismutase (E.C. 1.15.1.1), catalase (E.C. 1.11.1.6.), ascorbate peroxidase (E.C. 1.11.1.11)] were enhanced during the advanced phase of aging. The main non-enzymatic antioxidant compound, ascorbate, decreased markedly in the early stages of storage, followed by a slower decline. Total radical scavenging activity was also maintained at the end of the storage period. Our results indicate that the enhanced aging process occurring during storage at room temperature does not seem to be associated with the changes classically encountered during leaf senescence or seed aging and that the observed degenerative processes do not surpass the protective potential of the tubers.

The Dynamics of the Oxidant-Antioxidant Balance in the Early Phase of Human Acute Biliary Pancreatitis

Background/Aim: Reactive oxygen species play an important role in the pathogenesis of acute pancreatitis (AP) in animal models. Data on the oxidant-antioxidant balance in humans are scanty. The present study was undertaken to evaluate the dynamics of changes in the oxidant-antioxidant balance in the early phase of human AP. Methods: 74 consecutive patients with acute biliary pancreatitis (16 with severe, 58 with mild pancreatitis), treated endoscopically, were included in the study. Serum concentrations of sulfhydryl groups (SH; main nonenzymatic antioxidant; 73 patients) and thiobarbituric acid reactive substances (TBARS; markers of reactive oxygen species-mediated tissue damage; 56 patients) were determined on admission and on each of 10 successive days. The analysis comprised the comparison of results in patients with mild and severe outcome of pancreatitis. Results: Serum SH dropped by 27%, reaching the trough level on day 4 of hospitalization, whereas serum TBARS rose by 28%, reaching a peak 1 day later. Neither SH nor TBARS returned to initial values at the end of observation. The most dynamic changes in both SH and TBARS concentrations occurred in the first 3 days of hospitalization. The changes were significantly greater in patients with complicated pancreatitis in comparison to patients with mild disease, and were most pronounced in patients who developed infected pancreatic necrosis and who subsequently died. Conclusions: The oxidant-antioxidant balance changes rapidly in the early phase of human AP, confirming the role of oxidative stress in the pathogenesis of AP. The degree of changes correlates with the clinical severity of pancreatitis.