Blood glutathione homeostasis as a determinant of resting and exercise-induced oxidative stress in young men

Abstract

Although the importance of glutathione in protection against oxidative stress is well recognised, the role of physiological levels of glutathione and other endogenous antioxidants in protecting against exercise-induced oxidative stress is less clear. We evaluated the role of glutathione and selected antioxidant enzymes as determinants of lipid peroxidation at rest and in response to exercise in men (n = 13–14) aged 20–30 years, who cycled for 40 min at 60% of their maximal oxygen consumption (VO_2max). Levels of plasma thiobarbituric acid reactive substances (plasma TBARS) and blood oxidised glutathione (GSSG) increased by about 50% in response to exercise. Mean blood reduced glutathione (GSH)decreased by 13% with exercise. Of the measured red blood cell (RBC)antioxidant enzyme activities, only selenium-dependent glutathione peroxidase (Se-GPX) activity rose following exercise. In univariate regression analysis, plasma TBARS levels at rest predicted postexercise plasma TBARS and the exercise-induced change in total glutathione (TGSH). Blood GSSG levels at rest were strongly determinant of postexercise levels. Multiple regression analysis showed blood GSH to be a determinant of plasma TBARS at rest. The relative changes in TGSH were determinant of postexercise plasma TBARS. In summary, higher blood GSH and lower plasma TBARS at rest were associated with lower resting, and exercise-induced, lipid peroxidation. Subjects with a favourable blood glutathione redox status at rest maintained a more favourable redox status in response to exercise-induced oxidative stress. Changes in blood GSH and TGSH in response to exercise were closely associated with both resting and exercise-induced plasma lipid peroxidation. These results underscore the critical role of glutathione homeostasis in modulating exercise-induced oxidative stress and, conversely, the effect of oxidative stress at rest on exercise-induced changes in glutathione redox status.     

Glutathione metabolizing enzymes and oxidative stress in ferric nitrilotriacetate mediated hepatic injury

Summary

Glutathione (GSH) plays several important roles in the protection of cells against oxidative damage, particularly following exposure to xenobiotics. Ferric nitrilotriacetate (Fe-NTA) is a potent depletor of GSH and also enhances tissue lipid peroxidation. In this study, we show the effect of Fe-NTA treatment on hepatic GSH and some of the glutathione metabolizing enzymes, oxidant generation and liver damage. The level of hepatic GSH and the activities of glutathione reductase, glutathione S-transferase, glutathione peroxidase, and glucose 6-phosphate dehydrogenase all decrease following Fe-NTA administration. In these parameters the maximum decrease occurred at 12 h following Fe-NTA treatment. In contrast, γ-glutamyl transpeptidase was increased at this time. Not surprisingly, the increase in the activity of γ-glutamyl transpeptidase and decreases in GSH, glutathione peroxidase, glutathione reductase, glucose 6-phosphate dehydrogenase and glutathione S-transferase were found to be dependent on the dose of Fe-NTA administered. Fe-NTA administration also enhances the production of H₂O₂ and increases hepatic lipid peroxidation. Parallel to these changes, Fe-NTA enhances liver damage as evidenced by increases in serum transaminases. Once again, the liver damage is dependent on the dose of Fe-NTA and is maximal at 12 h. Pretreatment of animals with antioxidant, butylated hydroxy anisole (BHA), protects against Fe-NTA-mediated hepatotoxicity further supporting the involvement of oxidative stress in Fe-NTA-mediated hepatic damage. In aggregate, our results indicate that Fe-NTA administration eventuates in decreased hepatic GSH, a fall in the activities of glutathione metabolizing enzymes and excessive production of oxidants, all of which are involved in the cascade of events leading to iron-mediated hepatic injury.     

Riboflavin spraying impairs the antioxidant defense system but induces waterlogging tolerance in tobacco

Riboflavin, which causes plants to produce reactive oxygen species (ROS) when exposed to light, is an excellent photosensitizer for biocidal reactions. This study explores the possible protective role of riboflavin against waterlogging stress in tobacco plants. Tobacco seedlings (4 weeks old) were divided into four groups and pretreated with 0, 0.2, 0.5 or 1.0 mM riboflavin for 1 week, after which all groups were exposed to waterlogging stress for 7 days. We observed delayed leaf senescence and extended survival time, suggesting that riboflavin can confer increased waterlogging tolerance to plants as compared with the control (0 mM riboflavin). Enhanced stomatal closure was observed in the riboflavin-pretreated tobacco. We evaluated the levels of oxidative damage (H₂O₂ and lipid peroxidation), antioxidant enzyme (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) activity and antioxidant metabolites (including ascorbate and glutathione) in tobacco leaves that were pretreated with riboflavin. However, the results show that riboflavin pretreatment caused a decrease in chlorophyll content, antioxidant enzyme activity and redox values (AsA/DHA and GSH/GSSG), while causing a significant increase in lipid peroxidation, H₂O₂ accumulation and total ascorbate or glutathione content. In addition, the survival time and stomatal aperture of riboflavin-treated plants were significantly modified by exogenous application of GSH, well-known ROS scavenger. To explain the stomatal closure observed in tobacco plants, we propose a “damage avoidance” hypothesis based on riboflavin-mediated ROS toxicity. The protective function of the photosensitizer riboflavin may be highly significant for farming in frequently waterlogged areas.     

Survival of glioblastoma cells in response to endogenous and exogenous oxidative challenges: possible implication of NMDA receptor‐mediated regulation of redox homeostasis

Cancer cells are highly metabolically active and produce high levels of reactive oxygen species (ROS). Drug resistance in cancer cells is closely related to their redox status. The role of ROS and its impact on cancer cell survival seems far from elucidation. The mechanisms through which glioblastoma cells overcome aberrant ROS and oxidative stress in a milieu of hypermetabolic state is still elusive. We hypothesize that the formidable growth potential of glioma cells is through manipulation of tumor microenvironment for its survival and growth, which can be attributed to an astute redox regulation through a nexus between activation of N‐methyl‐d ‐aspartate receptor (NMDAR) and glutathione (GSH)‐based antioxidant prowess. Hence, we examined the NMDAR activation on intracellular ROS level, and cell viability on exposure to hydrogen peroxide (H₂O₂), and antioxidants in glutamate‐rich microenvironment of glioblastoma. The activation of NMDAR attenuated the intracellular ROS production in LN18 and U251MG glioma cells. MK‐801 significantly reversed this effect. On evaluation of GSH redox cycle in these cells, the level of reduced GSH and glutathione reductase (GR) activity were significantly increased. NMDAR significantly enhanced the cell viability in LN18 and U251MG glioblastoma cells, by attenuating exogenous H₂O₂‐induced oxidative stress, and significantly increased catalase activity, the key antioxidant that detoxifies H₂O₂. We hereby report an unexplored role of NMDAR activation induced protection of the rapidly multiplying glioblastoma cells against both endogenous ROS as well as exogenous oxidative challenges. We propose potentiation of reduced GSH, GR, and catalase in glioblastoma cells through NMDAR as a novel rationale of chemoresistance in glioblastoma.     

Preliminary studies on the involvement of glutathione metabolism and redox status against zinc toxicity in radish seedlings by 28-Homobrassinolide

The effect of exogenous application of 28-Homobrassinolide (HBR) on radish (Raphanus sativus L.) seedlings under zinc (Zn²⁺) stress on glutathione (GSH) production, consumption and changes in redox status was investigated. Zinc toxicity resulted in oxidative burst as evidenced by increased accumulation of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA) content. These stress indices were significantly decreased by HBR supplementation. Under Zn²⁺ stress, GSH pool was decreased, while the contribution of oxidized glutathione (GSSG) to total GSH increased (GSSH/GSH ratio), this translated into significant reduction of GSH redox homeostasis. In addition, an increase of phytochelatins (PCs) was observed. In radish seedlings under Zn²⁺ stress, the activities of gamma-glutamylcysteine synthetase (γ-ECS), glutathione synthetase (GS), glutathione peroxidase (GPX), glutathione-S-transferase (GST) and cysteine (Cys) levels increased but the activity of glutathione reductase (GR) decreased. However, application of HBR increased the GSH pool and maintained their redox ratio by increasing the enzyme activities of GSH biosynthesis (γ-ECS and GS) and GSH metabolism (GR, GPX and GST). The results of present study are novel in being the first to demonstrate that exogenous application of HBR modulates the GSH synthesis, metabolism and redox homeostasis to confer resistance against Zn²⁺ induced oxidative stress.     

Glutathione–ascorbic acid redox cycle and thioredoxin reductase activity in the digestive tract of Leptinotarsa decemlineata (Say)

In view of the antioxidant role of glutathione (GSH) and ascorbic acid (AA), we have examined capacity of the GSH–AA redox cycle in relation to oxidative stress effects in the midgut of the Colorado potato beetle Leptinotarsa decemlineata. Adult gut harbors a higher capacity to cope with oxidative stress than the larval gut. Protein carbonylation was pronounced in the wall of anterior larval midgut and was generally lower in the food digest than in the gut wall. Restriction of oxidative stress effects in anterior gut lumen manifested by lipid peroxidation and protein carbonylation is interpreted as a mechanism favoring digestion and absorption in the posterior midgut. Presence of high GSH in the posterior midgut and AA in both posterior and anterior midguts of adults points to higher utility of the GSH–AA redox system in limiting oxidative stress to manageable levels. The presence, gene expression and activity of thioredoxin reductase (TrxR) were demonstrated for the first time in L. decemlineata which was markedly higher in the anterior than in the posterior midgut in both stages. It is probably central to the maintenance of reduced GSH levels in the whole gut, despite a GSSG/2GSH redox potential tending towards oxidizing ranging from ?183.5 to ?124.4 mV. Glutathione-dehydroascorbate reductase (G_DHAR) activity was markedly augmented in adult gut compared with larva, pointing to a more efficient conversion of dehydroascorbate (DHA) to AA. Also, ascorbate peroxidase (APOX) activity was significantly elevated in all gut compartments of adult except the wall of posterior midgut. The results emphasize the potential importance and role of the GSH–AA redox cycle as a defense strategy against oxidative stress in the gut of L. decemlineata.     

Redox and nitric oxide homeostasis are affected in tomato (Solanum lycopersicum) roots under salinity-induced oxidative stress

The nicotinamide adenine dinucleotide phosphate (NADPH) and reduced glutathione (GSH) molecules play important roles in the redox homeostasis of plant cells. Using tomato (Solanum lycopersicum) plants grown with 120 mM NaCl, we studied the redox state of NADPH and GSH as well as ascorbate, nitric oxide (NO) and S-nitrosoglutathione (GSNO) content and the activity of the principal enzymes involved in the metabolism of these molecules in roots. Salinity caused a significant reduction in growth parameters and an increase in oxidative parameters such as lipid peroxidation and protein oxidation. Salinity also led to an overall decrease in the content of these redox molecules and in the enzymatic activities of the main NADPH-generating dehydrogenases, S-nitrosoglutathione reductase and catalase. However, NO content as well as gluthahione reductase and glutathione peroxidase activity increased under salinity stress. These findings indicate that salinity drastically affects redox and NO homeostasis in tomato roots. In our view, these molecules, which show the interaction between ROS and RNS metabolisms, could be excellent parameters for evaluating the physiological conditions of plants under adverse stress conditions.     

Ethylene reverses photosynthetic inhibition by nickel and zinc in mustard through changes in PS II activity,photosynthetic nitrogen use efficiency,and antioxidant metabolism

We investigated the influence of exogenously sourced ethylene (200 μL L^?1 ethephon) in the protection of photosynthesis against 200 mg kg^?1 soil each of nickel (Ni)- and zinc (Zn)-accrued stress in mustard (Brassica juncea L.). Plants grown with Ni or Zn but without ethephon exhibited increased activity of 1-aminocyclopropane carboxylic acid synthase, and ethylene with increased oxidative stress measured as H₂O₂ content and lipid peroxidation compared with control plants. The oxidative stress in Ni-grown plants was higher than Zn-grown plants. Under metal stress, ethylene protected photosynthetic potential by efficient PS II activity and through increased activity of ribulose-1,5-bisphosphate carboxylase and photosynthetic nitrogen use efficiency (P-NUE). Application of 200 μL L^?1 ethephon to Ni- or Zn-grown plants significantly alleviated toxicity and reduced the oxidative stress to a greater extent together with the improved net photosynthesis due to induced activity of ascorbate peroxidase and glutathione (GSH) reductase, resulting in increased production of reduced GSH. Ethylene formation resulting from ethephon application alleviated Ni and Zn stress by reducing oxidative stress caused by stress ethylene production and maintained increased GSH pool. The involvement of ethylene in reversal of photosynthetic inhibition by Ni and Zn stress was related to the changes in PS II activity, P-NUE, and antioxidant capacity was confirmed using ethylene action inhibitor, norbornadiene.     

Exogenous sodium nitroprusside and glutathione alleviate copper toxicity by reducing copper uptake and oxidative damage in rice (Oryza sativa L.) seedlings

Nitric oxide (NO) and glutathione (GSH) regulate a variety of physiological processes and stress responses; however, their involvement in mitigating Cu toxicity in plants has not been extensively studied. This study investigated the interactive effect of exogenous sodium nitroprusside (SNP) and GSH on Cu homeostasis and Cu-induced oxidative damage in rice seedlings. Hydroponically grown 12-day-old seedlings were subjected to 100 μM CuSO₄ alone and in combination with 200 μM SNP (an NO donor) and 200 μM GSH. Cu exposure for 48 h resulted in toxicity symptoms such as stunted growth, chlorosis, and rolling in leaves. Cu toxicity was also manifested by a sharp increase in lipoxygenase (LOX) activity, lipid peroxidation (MDA), hydrogen peroxide (H₂O₂), proline (Pro) content, and rapid reductions in biomass, chlorophyll (Chl), and relative water content (RWC). Cu-caused oxidative stress was evident by overaccumulation of reactive oxygen species (ROS; superoxide (O₂ ^?–) and H₂O₂). Ascorbate (AsA) content decreased while GSH and phytochelatin (PC) content increased significantly in Cu-stressed seedlings. Exogenous SNP, GSH, or SNP?+?GSH decreased toxicity symptoms and diminished a Cu-induced increase in LOX activity, O₂ ^?–, H₂O₂, MDA, and Pro content. They also counteracted a Cu-induced increase in superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), and glyoxalase I and glyoxalase II activities, which paralleled changes in ROS and MDA levels. These seedlings also showed a significant increase in catalase (CAT), glutathione peroxidase (GPX), dehydroascorbate reductase (DHAR), glutathione S-transferase (GST) activities, and AsA and PC content compared with the seedlings stressed with Cu alone. Cu analysis revealed that SNP and GSH restricted the accumulation of Cu in the roots and leaves of Cu-stressed seedlings. Our results suggest that Cu exposure provoked an oxidative burden while reduced Cu uptake and modulating the antioxidant defense and glyoxalase systems by adding SNP and GSH play an important role in alleviating Cu toxicity. Furthermore, the protective action of GSH and SNP?+?GSH was more efficient than SNP alone.     

Hydrogen peroxide-induced oxidative damages in <Emphasis Type="Italic">Schizosaccharomyces pombe</Emphasis>

Oxidative stress causes damage to proteins, lipids and nucleic acids, and thereby compromises cell viability. Some of the oxidative stress markers in an eukaryotic model organism, fission yeast Schizosaccharomyces pombe, were evaluated in this study. Intracellular oxidation, protein carbonyls, lipid peroxidation and reduced glutathione (GSH) levels were investigated in H₂O₂-treated and non-treated control cells. It was observed that increased H₂O₂ concentration proportionally lowered the cell number and increased the intracellular oxidation, lipid peroxidation and protein carbonyl levels in S. pombe. A dose-dependent decrease in GSH level was also detected. The fission yeast S. pombe is best known for its contribution to understanding of eukaryotic cell cycle control. S. pombe displays a different physiology from Saccharomyces cerevisiae in several ways and is thus probably more closely related to higher eukaryotes. The purpose of this study was to provide some data about the effects of hydrogen peroxide on the proteins and lipids in the fission yeast. The data obtained here is expected to constitute a basis for the further studies on redox balance and related processes in yeast and mammalian cells.     

Blood glutathione homeostasis as a determinant of resting and exercise-induced oxidative stress in young men

Although the importance of glutathione in protection against oxidative stress is well recognized, the role of physiological levels of glutathione and other endogenous antioxidants in protecting against exercise-induced oxidative stress is less clear. We evaluated the role of glutathione and selected antioxidant enzymes as determinants of lipid peroxidation at rest and in response to exercise in men (n = 13-14) aged 20-30 years, who cycled for 40 min at 60% of their maximal oxygen consumption (VO2max). Levels of plasma thiobarbituric acid reactive substances (plasma TBARS) and blood oxidised glutathione (GSSG) increased by about 50% in response to exercise. Mean blood reduced glutathione (GSH) decreased by 13% with exercise. Of the measured red blood cell (RBC) antioxidant enzyme activities, only selenium-dependent glutathione peroxidase (Se-GPX) activity rose following exercise. In univariate regression analysis, plasma TBARS levels at rest predicted postexercise plasma TBARS and the exercise-induced change in total glutathione (TGSH). Blood GSSG levels at rest were strongly determinant of postexercise levels. Multiple regression analysis showed blood GSH to be a determinant of plasma TBARS at rest. The relative changes in TGSH were determinant of postexercise plasma TBARS. In summary, higher blood GSH and lower plasma TBARS at rest were associated with lower resting, and exercise-induced, lipid peroxidation. Subjects with a favourable blood glutathione redox status at rest maintained a more favourable redox status in response to exercise-induced oxidative stress. Changes in blood GSH and TGSH in response to exercise were closely associated with both resting and exercise-induced plasma lipid peroxidation. These results underscore the critical role of glutathione homeostasis in modulating exercise-induced oxidative stress and, conversely, the effect of oxidative stress at rest on exercise-induced changes in glutathione redox status.     

Ameliorative Role of Castasterone on Copper Metal Toxicity by Improving Redox Homeostasis in <Emphasis Type="Italic">Brassica juncea</Emphasis> L.

The transition metal elements like copper act as double-edged sword for living cells. Cu, a redox active metal, is essential for various biological processes, but at higher concentrations it leads to toxicity by inducing production of reactive oxygen species (ROS). Thus, the objective of the present study was to investigate the effects of exogenously applied castasterone on oxidative stress markers and redox homeostasis managers in Brassica juncea plants subject to copper stress for 30 days. Copper-exposed plants showed accumulation of free radicals (H₂O₂ and superoxide anion) and lipid peroxidation. However, the exogenous treatment of seeds via the seed soaking method with different concentrations of castasterone reduced H₂O₂ production, superoxide anion radical content, and lipid peroxidation, thus indicating improved detoxification of ROS. Enzyme activity was increased by 19.19% for guaiacol peroxidase, 16.20% for superoxide dismutase, 35.74% for glutathione peroxidase, 27.58% for dehydroascorbate reductase, and 42.75% for ascorbate peroxidase, with castasterone pre-soaking under copper stress. The levels of non-enzymatic antioxidants were also increased with castasterone pre-treatment under copper stress. It may be concluded that castasterone treatment enhanced redox homeostasis managers in addition to increased levels of osmoprotectants.     

Toxicity of Fe2O3 nanoparticles on human blood lymphocytes

Magnetic nanoparticles (NPs) are used to a large extent in the targeted delivery of therapeutic agents. In this study, we aimed to investigate the possible toxicity of Fe₂O ₃ NPs on human cells, including blood lymphocytes. We isolated blood lymphocytes from healthy humans using Ficoll polysaccharide and subsequently by gradient centrifugation. Then, the toxicity parameters, including cell viability, reactive oxygen species (ROS) formation, lipid peroxidation, cellular glutathione (GSH) level, mitochondrial and lysosomal damage, were measured in blood lymphocytes after exposure to Fe ₂O ₃ NPs. Our results indicated that Fe ₂O ₃ NPs significantly (dependent on concentration) reduced the cell viability, and the IC ₅₀ was determined to be 1 mM. With increasing concentrations, we found that Fe ₂O ₃ NPs–induced cell toxicity was associated with a significant increase in intracellular ROS and loss of mitochondrial membrane potential and lysosomal membrane leakiness. Consequently, these NPs at different concentrations affect GSH level and cause oxidative stress in human lymphocytes.     

Antioxidant and hepatoprotective potential of Aegle marmelos Correa. against CCl4-induced oxidative stress and early tumor events

The antioxidant properties and inhibitory effect on early tumor promoter markers of A. marmelos (25 and 50 mg/Kg b. wt. orally) have been evaluated. Male Wistar rats were pre-treated for seven consecutive days with A. marmelos prior to CCl₄ (1 mL Kg^{? 1} body weight p. o., in corn oil [1:1 v/v]) treatment. Pre-treatment with A. marmelos suppressed lipid peroxidation (LPO), xanthine oxidase (XO) and release of serum toxicity marker enzymes viz, SGOT, LDH, SGPT dose-dependently and significantly (p < 0.001). Hepatic antioxidant status viz, reduced glutathione (GSH), glutathione reductase (GR), glutathione peroxidase (GPx), quinone reductase (QR), catalase (CAT) were concomitantly restored in A. marmelos-treated groups (p < 0.001). In addition, A. marmelos pretreatment also prevented the CCl₄-enhanced ornithine decarboxylase (ODC) and hepatic DNA synthesis significantly (p < 0.001). In conclusion, carbon tetrachloride-induced liver toxicity was strikingly attenuated by A. marmelos treatment and the study gives some insight into the mechanisms involved in diminution of free radical generating toxicants and enhancement of the antioxidant armory, hence preventing further tissue damage, injury and hyperproliferation.

Thus, these findings indicate that A. marmelos attenuates CCl₄-mediated hepatic oxidative stress, toxicity, tumor promotion and subsequent cell proliferation response in Wistar rats.     

Cobalt-induced oxidative stress causes growth inhibition associated with enhanced lipid peroxidation and activates antioxidant responses in Indian mustard (Brassica juncea L.) leaves

The effect of 100 μM cobalt (Co) on plant growth and on biochemical parameters indicative of oxidative stress was investigated in a hydroponic experiment. The responses of antioxidant enzymes and compounds of the ascorbate–glutathione (AsA–GSH) cycle were also assessed on the hyperaccumulating plant, Indian mustard (Brasssica juncea L.). The effect of excess Co was associated with a significant increase in the levels of proline, carbonylated protein, malondialdehyde, superoxide anion (O ₂ ^·? ), and hydrogen peroxide (H₂O₂), and resulted in the accumulation of Co. Co toxicity was associated with an increase in the volume of palisade and spongy cells, and a reduction in the number of chloroplasts per cell. Co-induced cell death was characterized by DNA fragmentation and a 36 kDa DNase activity. Despite decreased catalase activity, peroxidase, superoxide dismutase, and AsA–GSH cycle-related enzymes including monodehydroascorbate reductase, dehydroascorbate reductase, and glutathione reductase exhibited remarkable induction under Co stress. Furthermore, the contents of reduced and oxidized forms of ascorbate and glutathione were significantly increased with Co supplementation. Co treatment led to the activation of 44 and 46 kDa mitogen-activated protein kinase (MAPK) and indicated the role of the MAPK cascade in transducing Co-mediated signals. The present results suggest that excess Co reduces seedling growth by inducing oxidative stress related to lipid peroxidation and overproduction of O ₂ ^·? and H₂O₂. The stimulated activities of antioxidative enzymes and induction of MAPKs did not reverse the oxidative stress caused by Co-induced reactive oxygen species generation in Indian mustard seedlings.     

Cadmium causes oxidative stress in mung bean by affecting the antioxidant enzyme system and ascorbate-glutathione cycle metabolism

Ascorbate (AsA)-glutathione (GSH) cycle metabolism is an essential mechanism for the resistance of plants under stress conditions. In a greenhouse pot experiment, the influence of cadmium (Cd) (25, 50, and 100 mg/kg soil) on plant dry weight and leaf area, photosynthetic parameters (net photosynthetic rate (P_N) and chlorophyll (Chl) content) and oxidative stress, and the possible protective role of AsA-GSH cycle metabolism was studied in two mung bean (Vigna radiata (L.) Wilczek.) cvs. Pusa 9531 (Cd-tolerant) and PS 16 (Cd-susceptible) at 30 days after sowing. The contents of thiobarbituric acid-reactive substances (TBARS), H₂O₂, and the leakage of ions were the highest at 100 mg Cd/kg soil, and the effect was more pronounced in cv. PS 16 than in cv. Pusa 9531. This was concomitant with the strongest decreases in P_N, plant dry weight, and leaf area. The changes in the AsA-GSH redox state and an increase in AsA-GSH-regenerating enzymes, such as glutathione reductase, monodehydroascorbate reductase, dehydroascorbate reductase, and other antioxidant enzymes, such as superoxide dismutase and ascorbate peroxidase, strongly supported over-utilization of AsA-GSH in Cd-treated plants. However, the oxidative stress caused by Cd toxicity was partially overcome by AsA-GSH-based detoxification mechanism in the two genotypes studied because an increases in lipid peroxidation (TBARS, ion leakage) and H₂O₂ content were accompanied by a corresponding decrease in reduced AsA and GSH pools. Thus, changes in AsA-GSH pools and the coordination between AsA-GSH-regenerating enzymes and other enzymatic antioxidants of the leaves suggest their relevance to the defense against Cd stress.     

The effect of post-hypoxia on roots inSenecio andMyosotis species related to the glutathione system

This paper shows the effect of re-aeration following hypoxic pretreatment on the glutathione system in plants with different flooding tolerance. Re-aeration of hypoxically pretreated roots led to an increase of TBA-rm content indicating an accelerated lipid peroxidation (post-anoxic injury). Re-admission of oxygen resulted in a clear increase in the content of total glutathione in both flooding-intolerant speciesMyosotis arvensis andSenecio jacobaea. Simultaneously, the high ratio between reduced (GSH) and oxidized (GSSG) glutathione decreased in these species upon the onset of re-aeration, while the tolerantMyosotis palustris andSenecio aquaticus showed only little changes in contents of GSH and GSSG. An imbalance in GSH/GSSG ratio reflects oxidative stress. The glutathione reductase (GR) reacted very differently in the investigated genera. The metabolic response to varying oxygen pressure is much stronger in the flooding-intolerant species compared to species naturally growing in wetlands. The present results suggest that glutathione system is an important component in overcoming oxidative stress.     

Arsenite treatment induces oxidative stress,upregulates antioxidant system,and causes phytochelatin synthesis in rice seedlings

The effects of arsenite treatment on generation of reactive oxygen species, induction of oxidative stress, response of antioxidative system, and synthesis of phytochelatins were investigated in two indica rice (Oryza sativa L.) cvs. Malviya-36 and Pant-12 grown in sand cultures for a period of 5–20 days. Arsenite (As₂O₃; 25 and 50 μM) treatment resulted in increased formation of superoxide anion (O₂^.−), elevated levels of H₂O₂ and thiobarbituric acid reactive substances, showing enhanced lipid peroxidation. An enhanced level of ascorbate (AA) and glutathione (GSH) was observed irrespective of the variation in the level of dehydroascorbate (DHA) and oxidized glutathione (GSSG) which in turn influenced redox ratios AA/DHA and GSH/GSSG. With progressive arsenite treatment, synthesis of total acid soluble thiols and phytochelatins (PC) increased in the seedlings. Among antioxidative enzymes, the activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6), total ascorbate peroxidase (APX, EC 1.11.1.11), chloroplastic ascorbate peroxidase, guaiacol peroxidase (EC 1.11.1.7), monodehydroascorbate reductase (EC 1.6.5.4), and glutathione reductase (EC 1.6.4.2) increased in arsenite treated seedlings, while dehyroascorbate reductase (EC 1.8.5.1) activity declined initially during 5–10 days and increased thereafter. Results suggest that arsenite treatment causes oxidative stress in rice seedlings, increases the levels of many enzymatic and non-enzymatic antioxidants, and induces synthesis of thiols and PCs, which may serve as important components in mitigating arsenite-induced oxidative damage.     

N-acetylcysteine amide,a thiol antioxidant,prevents bleomycin-induced toxicity in human alveolar basal epithelial cells (A549)

Abstract

Bleomycin (BLM), a glycopeptide antibiotic from Streptomyces verticillus, is an effective antineoplastic drug. However, its clinical use is restricted due to the wide range of associated toxicities, especially pulmonary toxicity. Oxidative stress has been implicated as an important factor in the development of BLM-induced pulmonary toxicity. Previous studies have indicated disruption of thiol-redox status in lungs (lung epithelial cells) upon BLM treatment. Therefore, this study focused on (1) investigating the oxidative effects of BLM on lung epithelial cells (A549) and (2) elucidating whether a well-known thiol antioxidant, N-acetylcysteine amide (NACA), provides any protection against BLM-induced toxicity. Oxidative stress parameters, such as glutathione (GSH), malondialdehyde (MDA), and antioxidant enzyme activities were altered upon BLM treatment. Loss of mitochondrial membrane potential (ΔΨm), as assessed by fluorescence microscopy, indicated that cytotoxicity is possibly mediated through mitochondrial dysfunction. Pretreatment with NACA reversed the oxidative effects of BLM. NACA decreased the reactive oxygen species (ROS) and MDA levels and restored the intracellular GSH levels. Our data showed that BLM induced A549 cell death by a mechanism involving oxidative stress and mitochondrial dysfunction. NACA had a protective role against BLM-induced toxicity by inhibiting lipid peroxidation, scavenging ROS, and preserving intracellular GSH and ΔΨm. NACA can potentially be developed into a promising adjunctive therapeutic option for patients undergoing chemotherapy with BLM.