Metal-induced Oxidative Stress Research Articles

Proteomic responses to metal-induced oxidative stress in hydrothermal vent-living mussels, Bathymodiolus sp., on the Southwest Indian Ridge

Bathymodiolin mussels are amongst the dominant fauna occupying hydrothermal vent ecosystems throughout the World's oceans. This subfamily inhabits a highly ephemeral and variable environment, where exceptionally high concentrations of reduced sulphur species and heavy metals necessitate adaptation of specialised detoxification mechanisms. Whilst cellular responses to common anthropogenic pollutants are well-studied in shallow-water species, they remain limited in deep-sea vent fauna. Bathymodiolus sp. were sampled from two newly-discovered vent sites on the Southwest Indian Ridge (Tiamat and Knuckers Gaff) by the remotely operated vehicle (ROV) Kiel 6000 during the RRS James Cook cruise, JC 067 in November 2011. Here, we use redox proteomics to investigate the effects of tissue metal accumulation on protein expression and thiol oxidation in gill. Following 2D PAGE, we demonstrate a significant difference in intensity in 30 protein spots in this organ between the two vent sites out of 205 matched spots. We also see significant variations in thiol oxidation in 15 spots, out of 143 matched. At Tiamat, 23 protein spots are up-regulated compared to Knuckers Gaff and we identify 5 of these with important roles in metabolism, cell structure, stress response, and redox homeostasis. We suggest that increased metal exposure triggers changes in the proteome, regulating tissue uptake. This is evident both between vent sites and across a chemical gradient within the Knuckers Gaff vent site. Our findings highlight the importance of proteomic plasticity in successful adaptation to the spatially and temporally fluctuating chemical environments that are characteristic of hydrothermal vent habitats.

Metal-Induced Oxidative Stress in Egyptian Women with Breast Cancer

Metal-Induced Oxidative Stress in Egyptian Women with Breast Cancer

Glutathione is a key player in metal-induced oxidative stress defenses.

Since the industrial revolution, the production, and consequently the emission of metals, has increased exponentially, overwhelming the natural cycles of metals in many ecosystems. Metals display a diverse array of physico-chemical properties such as essential versus non-essential and redox-active versus non-redox-active. In general, all metals can lead to toxicity and oxidative stress when taken up in excessive amounts, imposing a serious threat to the environment and human health. In order to cope with different kinds of metals, plants possess defense strategies in which glutathione (GSH; γ-glu-cys-gly) plays a central role as chelating agent, antioxidant and signaling component. Therefore, this review highlights the role of GSH in: (1) metal homeostasis; (2) antioxidative defense; and (3) signal transduction under metal stress. The diverse functions of GSH originate from the sulfhydryl group in cysteine, enabling GSH to chelate metals and participate in redox cycling.

Cadmium accumulation and strategies to avoid its toxicity in roots of the citrus rootstock Citrumelo

In order to assess implications of Cd-induced oxidative stress in roots of the citrus rootstock Citrumelo, seedlings were hydroponically exposed to two relatively realistic Cd concentrations during 7 days. Our results showed that increasing Cd concentrations in external solution were associated with higher Cd accumulations in roots. At 5μM Cd the accumulation of Cd in roots was over 70-f higher than in aerial part (stem+leaves). Malondialdehyde (MDA), superoxide radical (O2−), hydrogen peroxide (H2O2) and lipoxygenase activity (LOX) increased in Cd-exposed roots, suggesting a metal-induced oxidative stress. The Cd treatment enhanced the activity of antioxidant enzymes superoxide dismutase (SOD), catalase (CAT) and guaiacol-type peroxidase (G-POD), as well as the content of secondary metabolites i.e. soluble phenolics (SPs) and lignin. Histochemical analyses of roots showed that Cd, H2O2, (O2−), lignin and G-POD displayed a similar location pattern. Almost all analyzed parameters showed a similar dynamic tendency with increases under 5μM Cd followed by decreases under 10μM Cd, suggesting that a complex coordinated Cd-defensive mechanism is operating in Citrumelo roots exposed to environmental realistic Cd concentrations.

Transcription of genes involved in glutathione biosynthesis in the solitary tunicate Ciona intestinalis exposed to metals

Exposure to metals is known to generate oxidative stress risk in living organisms, which are able to respond with the induction of antioxidant defenses, both enzymatic and non-enzymatic. Glutathione (GSH) is considered to be an important cellular component involved in protecting cells, both as metal chelating agent and oxygen radical scavenger. In this work we used molecular techniques to analyze the nucleotide and predicted amino acid sequences of genes involved in GSH biosynthesis, γ-glutamyl-cysteine ligase catalytic subunit (ci-gclc), γ-glutamyl-cysteine ligase modifier subunit (ci-gclm) and GSH synthase (ci-gs) in the solitary tunicate Ciona intestinalis. We also studied the transcription of the above genes after in vivo exposure to Cd, Cu and Zn by semiquantitativ RT-PCR to improve our knowledge about the relationship between metal-induced oxidative stress and GSH production and locate mRNA expression by in situ hybridization (ISH). These genes exhibit a good level of sequence conservation with metazoan homologs generally, especially for residues important for the activity of the enzymes. Phylogenetic analyses indicate that the three enzymes evolved in different ways, Ci-GCLC and Ci-GS being mostly correlated with invertebrate proteins, Ci-GCLM being as sister group of vertebrate GCLMs. Our in silico analyses of the ci-gs and ci-gclc promoter regions revealed putative consensus sequences similar to mammalian metal-responsive elements (MRE) and antioxidant response elements (ARE), indicating that the transcription of these genes may directly depend on metals and/or reactive oxygen species. Results highlight a statistically significant increase in gene transcription, demonstrating that metal treatments have inducible effects on these genes. They can modulate gene transcription not only through MREs but also through AREs, as a consequence of metal-dependent ROS formation. The ISH location of Ci-GS and Ci-GCLC mRNAs shows that the cells most involved in glutathione biosynthesis are circulating hemocytes. The data presented here emphasize the importance of complex metal regulation of ci-gclc, ci-gclm and ci-gs transcription, which can create an efficient detoxification pathway allowing C. intestinalis to survive in continued elevated presence of metals in the environment.

Redox active metal-induced oxidative stress in biological systems

A number of studies performed on biological systems have shown that redox-active metals such as iron and copper as well as other transition metals can undergo redox cycling reactions and produce reactive free radicals termed also reactive oxygen species (ROS) or reactive nitrogen species (RNS). The most representative examples of ROS and RNS are the superoxide anion radical and nitric oxide, respectively, both playing a dual role in biological systems. At low/moderate concentrations of ROS and RNS, they can be involved in many physiological roles such as defense against infectious agents, involvement in a number of cellular signaling pathways and other important biological processes. On the other hand, at high concentrations, ROS and RNS can be important mediators of damage to biomolecules involving DNA, membrane lipids, and proteins. One of the most damaging ROS occurring in biological systems is the hydroxyl radical formed via the decomposition of hydrogen peroxide catalyzed by traces of iron, copper and other metals (the Fenton reaction). The hydroxyl radical is known to react with the DNA molecule, forming 8-OH-Guanine adduct, which is a good biomarker of oxidative stress of an organism and a potential biomarker of carcinogenesis. This review discusses the role of iron and copper in uncontrolled formation of ROS leading to various human diseases such as cancer, cardiovascular disease, and neurological disorders (Alzheimer’s disease and Parkinson’s disease). A discussion is devoted to the various protective antioxidant networks against the deleterious action of free radicals. Metal-chelation therapy, which is a modern pharmacotherapy used to chelate redox-active metals and remove toxic metals from living systems to avoid metal poisoning, is also discussed.

The Role of Iron Metabolism in Lung Inflammation and Injury.

Iron is required for many vital functions including oxygen transport and energy metabolism. Protective mechanisms maintain optimal iron concentration involving dynamic regulation of the transporters and iron storage proteins. In addition to these systemic regulatory mechanisms, the unique lung environment must provide detoxification from metal-induced oxidative stress and pathogenic infections. This review focuses on the unique role of iron metabolism in lung injury and inflammation.

Metal-Induced Oxidative Stress in Egyptian Women with Breast Cancer

Some trace metals are toxic and claimed to be carcinogenic. The present work aimed to determine the levels of some trace metals in breast tissues (healthy and tumor specimens) and to evaluate their concentrations in relation to the oxidative stress status in breast cancer which is a major health problem among Egyptian women. This study included 127 female patients with breast swelling. Analysis of trace metals in breast tissue was done using Atomic Absorption Spectrophotometry (AAS). Lipid peroxidation and oxidative status were assessed. There were statistically high levels of iron, zinc and copper in the benign and malignant breast tissues in comparison to the control group. Higher Malondialdehyde (MDA) levels were detected in patients with breast tumors while Superoxide dismutase (SOD), Catalase (CAT), Glutathione peroxidase (GSH), Glutathione-S-transferase (GST) and Nitric oxide (NO) were low in comparison to the healthy group. In conclusion, the alteration of the elemental content in cancerous breast tissues and the disruption of oxidant/antioxidant balance highlight the role of trace metals in cancer development.

Metal-Induced Oxidative Stress and Plant Mitochondria

A general status of oxidative stress in plants caused by exposure to elevated metal concentrations in the environment coincides with a constraint on mitochondrial electron transport, which enhances ROS accumulation at the mitochondrial level. As mitochondria are suggested to be involved in redox signaling under environmental stress conditions, mitochondrial ROS can initiate a signaling cascade mediating the overall stress response, i.e., damage versus adaptation. This review highlights our current understanding of metal-induced responses in plants, with focus on the production and detoxification of mitochondrial ROS. In addition, the potential involvement of retrograde signaling in these processes will be discussed.

Toxicity assessment of contaminated soils from a mining area in Northeast Italy by using lipid peroxidation assay

Contamination by heavy metals in soils may strongly affect the environmental quality. Lipid peroxidation caused by heavy metals in plants was investigated as a relevant bioassay of toxicity. Soils and wild plants (dandelion and willow) were collected from an abandoned mine area in northeast Italy, and the concentration of different heavy metals (Ni, Cr, Cu, Pb, Zn, Fe and Mn) were measured and analyzed. Soils affected by mining activities presented total Zn, Cu, and Pb concentrations (2566, 3975, 20,815mgkg−1 respectively) above toxic thresholds, and 58% for Fe. Heavy metal-induced oxidative stress was evidenced by the generation of reactive radicals, followed by an increase in malondialdehyde (MDA) production up to 41.64μM in willow leaves. We found that MDA concentration in plant tissues differed significantly among species and plant organs. The higher concentration of metal in soil corresponded with the higher concentration of MDA in the plant. The combined results of metal concentration, MDA content and translocation coefficients in plants show that the investigated plants are rather highly tolerant towards environmental pollution. This suggests that they could be useful in phytoremediation of metal contaminated sites.

Effects of Pb and Ni stress on oxidative stress parameters in three moss species

Antioxidative responses of the mosses Hypnum plumaeforme, Thuidium cymbifolium, and Brachythecium piligerum to short-term Pb and Ni stress were investigated. Both Pb and Ni treatment increased the formation of reactive oxygen species and lipid peroxidation, decreased superoxide dismutase (SOD), and catalase (CAT) activities in H. plumaeforme and T. cymbifolium. However, SOD activity in B. piligerum was increased under 10 mM Pb stress and Ni increased CAT activity in B. piligerum under 1 mM Ni stress. Peroxidase (POX) activity in the three mosses was increased by Pb and Ni exposure, indicating that POX plays an important role in preventing heavy metal-induced oxidative stress. The accumulation of O 2 − and H 2O 2 in mosses is related to the decline in SOD and CAT activities. B. piligerum is the most sensitive and T. cymbifolium is the most tolerant species to Pb and Ni stress among the three bryophytes.

Redox control of prion and disease pathogenesis.

Imbalance of brain metal homeostasis and associated oxidative stress by redox-active metals like iron and copper is an important trigger of neurotoxicity in several neurodegenerative conditions, including prion disorders. Whereas some reports attribute this to end-stage disease, others provide evidence for specific mechanisms leading to brain metal dyshomeostasis during disease progression. In prion disorders, imbalance of brain-iron homeostasis is observed before end-stage disease and worsens with disease progression, implicating iron-induced oxidative stress in disease pathogenesis. This is an unexpected observation, because the underlying cause of brain pathology in all prion disorders is PrP-scrapie (PrP(Sc)), a beta-sheet-rich conformation of a normal glycoprotein, the prion protein (PrP(C)). Whether brain-iron dyshomeostasis occurs because of gain of toxic function by PrP(Sc) or loss of normal function of PrP(C) remains unclear. In this review, we summarize available evidence suggesting the involvement of oxidative stress in prion-disease pathogenesis. Subsequently, we review the biology of PrP(C) to highlight its possible role in maintaining brain metal homeostasis during health and the contribution of PrP(Sc) in inducing brain metal imbalance with disease progression. Finally, we discuss possible therapeutic avenues directed at restoring brain metal homeostasis and alleviating metal-induced oxidative stress in prion disorders.

Cadmium induced-oxidative stress in pituitary gland is reversed by removing the contamination source

Cadmium (Cd(2+)) is one of the most important environmental contaminants and acts as an endocrine disruptor. Previously, we have demonstrated that the simultaneous administration of Cd(2+) and melatonin (Mel) in drinking water impaired metal-induced oxidative stress in rat anterior pituitary gland. The aim of this study was to investigate if a treatment started after the toxic manifestations of Cd( 2+) became evident could reverse the effects of the metal. Animals exposed to Cd(2+) (5 parts per million [ppm], 30 days) were treated with Mel or without the metal during the next 1 or 2 months. Cd(2+) exposure increased the expression of heme oxygenase-1 (HO-1), a biomarker of oxidative stress, and an a posteriori Mel treatment reversed oxidative stress induced by Cd(2+). This effect was also observed 1 month after metal removal. The Cd(2+)-induced increase in metallothionein-1 (MT-1) and nitric oxide synthase 1 (NOS1) expression were also reversed by metal removal. In addition, serum prolactin and luteinizing hormone levels affected by Cd( 2+) exposure were normalized. Considering that the manifestations of Cd(2+) intoxication become evident only after a certain period of metal accumulation, these results show that metal removal is enough to reverse Cd(2+) effects in anterior pituitary gland and bring to light the relevance of moving away the individual from the contamination source.

Inhibition of Prolyl Hydroxylase Protects against 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Neurotoxicity: MODEL FOR THE POTENTIAL INVOLVEMENT OF THE HYPOXIA-INDUCIBLE FACTOR PATHWAY IN PARKINSON DISEASE

Hypoxia-inducible factor (HIF) plays an important role in cell survival by regulating iron, antioxidant defense, and mitochondrial function. Pharmacological inhibitors of the iron-dependent enzyme class prolyl hydroxylases (PHD), which target alpha subunits of HIF proteins for degradation, have recently been demonstrated to alleviate neurodegeneration associated with stroke and hypoxic-ischemic injuries. Here we report that inhibition of PHD by 3,4-dihydroxybenzoate (DHB) protects against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced nigral dopaminergic cell loss and up-regulates HIF-1alpha within these neurons. Elevations in mRNA and protein levels of HIF-dependent genes heme oxygenase-1 (Ho-1) and manganese superoxide dismutase (Mnsod) following DHB pretreatment alone are also maintained in the presence of MPTP. MPTP-induced reductions in ferroportin and elevations in nigral and striatal iron levels were reverted to levels comparable with that of untreated controls with DHB pretreatment. Reductions in pyruvate dehydrogenase mRNA and activity resulting from MPTP were also found to be attenuated by DHB. In vitro, the HIF pathway was activated in N27 cells grown at 3% oxygen treated with either PHD inhibitors or an iron chelator. Concordant with our in vivo data, the MPP(+)-elicited increase in total iron as well as decreases in cell viability were attenuated in the presence of DHB. Taken together, these data suggest that protection against MPTP neurotoxicity may be mediated by alterations in iron homeostasis and defense against oxidative stress and mitochondrial dysfunction brought about by cellular HIF-1alpha induction. This study provides novel data extending the possible therapeutic utility of HIF induction to a Parkinson disease model of neurodegeneration, which may prove beneficial not only in this disorder itself but also in other diseases associated with metal-induced oxidative stress.

The effects of adrenomedullin and methylated‐adrenomedullin treatments on some antioxidative enzyme activities and malondialdehyde levels in rat liver

Adrenomedullin (AdM) has many important effects such as vasorelaxant, natriuretic, proliferative/antiproliferative, regulator of adrenal hormones and ACTH secretion, antiapoptotic and antioxidant. The aim of this study is to investigate the effects of AdM and methylated‐AdM (M‐AdM) on benzo(a)pyrene (B(a)p) and heavy metals (Cd, Pb, Hg) induced oxidative stress in the rat liver. For this purpose, at first AdM was methylated. Catalase (CAT), superoxide dismutase (SOD), glutation peroxidase (GSH‐Px), and MDA levels were assessed by spectrophotometrically. Administration of M‐AdM in the Pb‐treated rats significantly increased MDA levels compared with Pb‐treated group. While AdM further decreased Pb‐induced attenuation of SOD activity, MAdM did not change. However, MAdM significantly increased SOD activity compared with AdM in Pb‐treated rats. Hg‐induced decrease of SOD activity was reversed by M‐AdM administration significantly. The administration of AdM and MAdM in B(a)p‐treated rats increased SOD, CAT and GSH‐Px activities significantly compared with B(a)p‐treated group and the increases of antioxidant enzyme activities in MAdM group more than the AdM group. These results suggest that M‐AdM may have more potent protective effect than AdM against B(a)p or heavy metal‐induced oxidative stress (Supported by The Scientific and Technological Research Council of Turkey, Project No:106T112).

Research Highlights

Research Highlights

Metal-induced oxidative stress impacting plant growth in contaminated soil is alleviated by microbial siderophores

High levels of metals impede plant growth by affecting physiological processes. Siderophores are microbial Fe-chelators that, however, bind other metals. This study evaluated plant growth in a soil containing elevated levels of metals, including Al, Cu, Fe, Mn, Ni, and U, using Streptomyces-derived cell-free supernatant containing siderophores and auxins. Cowpea plants in the soil were treated with the culture filtrate. Growth was measured and biochemical analyses such as chlorophyll contents, RNA and protein quantification, lipid membrane peroxidation, and anti-oxidative responses were conducted to evaluate oxidative stress in the plants. Liquid chromatography–mass spectrometry was used to simulate competition for siderophore binding, and metal content of plants was determined spectroscopically. Whereas the metals inhibited plant growth, addition of siderophores improved growth. There was evidence of lipid peroxidation, an enhanced superoxide dismutase activity, and lowered chlorophyll, RNA, protein, carotenoid and residual indole acetic acid contents, especially in control plants. Siderophore competition assays between Al and Fe, and Fe and Cu suggested that trivalent metals are more competitive for siderophore binding than divalent ones. Compared to control plants, higher amounts of metals were obtained in siderophore-treated plants. Siderophores were able to supply plants with Fe in the presence of levels of metals, mainly Al, Cu, Mn, Ni and U that otherwise inhibit Fe acquisition. This led to enhanced chlorophyll content, circumventing lipid peroxidation effects on leaves. Siderophores lowered the formation of free radicals, thereby protecting microbial auxins from degradation and enabling them to enhance plant growth which in turn resulted in augmented metal uptake.

Aluminium and lead toxicity revisited: mechanisms explaining the particular sensitivity of the brain to oxidative damage

For several reasons the central nervous system is particularly sensitive to metal-induced oxidative stress (Verstraeten et al. 2008, this issue): the brain contains a high fraction of oxidizable polyunsaturated fatty acids, only relatively low activities of the antioxidant enzymes catalase, superoxide dismutase and glutathione peroxidase, has a high oxygen consumption and a high iron content. Parkinson’s disease is one of the neurodegenerative diseases where metal toxicity is suspected to play an important role (Verstraeten et al. 2008, this issue). This is based on the observation of high aluminium concentrations in post-mortem brains of patients with Parkinson’s disease and on animal studies where administration of aluminium caused a strong decrease in the dopamine content of the striatum. Metals are not degraded and may accumulate to toxic levels. This and the diversity of not yet fully understood mechanisms is the reason why metal toxicity is one of the hot topics of our journal (Hengstler et al. 2003; Barbosa et al. 2006a, b; Brulport et al. 2007; Devi et al. 2007; Hengstler and Bolt 2007; Kobayashi et al. 2007; M’Bemba-Meka et al. 2007; Montes et al. 2007; Nampoothiri et al. 2007; Periyakaruppan et al. 2007; Posser et al. 2007; Santos et al. 2007; Beyersmann and Hartwig 2008; Glahn et al. 2008). The editors are happy that Sandra Verstraeten from Buenos Aires and Lucila Aimo as well as Patricia Oteiza from the University of California present an extensive review about molecular mechanisms of brain toxicity in this issue of the Archives of Toxicology. The authors revisit the mechanisms of aluminium and lead toxicity and focus on the recent literature about deregulation of cell signalling, the impairment of neurotransmission, membrane biophysics and reactive oxygen species as key factors. This review is a must for everybody interested in metal-induced neurotoxicity.

The effect of Fenton reaction on protease-resistant prion protein (PrP Sc) degradation and scrapie infectivity

In prion diseases, metal imbalances in brain and/or metal substitutions for copper in prion protein suggest that metal-catalyzed oxidation (MCO) and oxidative stress may affect cellular function and accumulation of protease-resistant prion protein (PrP Sc). We examined the effect of metal-induced oxidative stress by Fenton reaction on prion protein with regard to its degradation, insolubility, and infectivity. Precipitation and insolubility of prion protein were induced by Fenton reaction in scrapie-infected brain homogenate. Results showed an increase in hydroxylation products (thiobarbituric acid reactive substances; TBARS) and a decrease of ferrous ion (Fe 2+) levels after Fenton reaction. Efficiency of metal-induced oxidation was higher for Fe 2+ than Mn 2+. Compared to untreated samples, there was increased susceptibility to proteolytic degradation of PrP Sc after treatment with 3.12–12.5 mM Fe 2+–Mn 2+/H 2O 2. Interestingly, we observed that Fenton reaction could extend incubation periods, indicating a decrease in scrapie infectivity. These results suggest that PrP Sc hydroxylation and degradation may affect PrP conversion and the pathogenesis of prion diseases.

The oxidation of glutathione by cobalt/tungsten carbide contributes to hard metal-induced oxidative stress

The occupational exposure to cobalt/tungsten carbide (Co/WC) dusts causes asthma and interstitial fibrosis. The International Agency for Research on Cancer (IARC) recently classified the mixture Co/WC as probably carcinogenic to humans (group 2A). The mechanism of action of Co/WC involves particle driven generation of Reactive Oxygen Species (ROS) with consequent oxidative damage. The present study evaluates the reactivity of Co/WC dust toward glutathione (GSH) and cysteine (Cys). Co/WC oxidized thiols through a mechanism involving the generation of sulphur-centred radicals. The results are consistent with the oxidation taking place at surface active sites, a part of which is accessible only to Cys S-H groups, but not to GSH ones. Such a reaction, with consequent irreversible depletion of antioxidant defenses of cells, will potentiate the oxidative stress caused by particle and cell generated ROS.