Role of hydrogen peroxide and oxidative stress in healing responses

Rojkind, Marcos; Domínguez-Rosales, José-Alfredo; Nieto, Natalia; Greenwel, Patricia

doi:10.1007/pl00012511

Cited by 218 publications

(167 citation statements)

References 227 publications

Supporting

Mentioning

153

Contrasting

Unclassified

Order By: Relevance

“…The plasmid encodes a fusion protein of the insert and an N-terminal extension containing a (His) 6 tag, enabling purification by nickel affinity chromatography. The construct was expressed in the JM109 strain of Escherichia coli.…”

Section: Methodsmentioning

confidence: 99%

“…Thus, changes in the redox equilibrium influence a host of cell functions, including but not limited to growth, stress responses, differentiation, metabolism, cell cycle, communication, migration, gene transcription, ion channels, and immune responses (for reviews see Refs. [2][3][4][5][6]. Alterations in the redox equilibrium are reflected in changes of the glutathione/glutathione-disulfide ratio (GSH/ GSSG) and the reduced/oxidized thioredoxin ratio.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Imaging Dynamic Redox Changes in Mammalian Cells with Green Fluorescent Protein Indicators

Dooley

Dore

Hanson

et al. 2004

Journal of Biological Chemistry

688

698

View full text Add to dashboard Cite

Changes in the redox equilibrium of cells influence a host of cell functions. Alterations in the redox equilibrium are precipitated by changing either the glutathione/glutathione-disulfide ratio (GSH/GSSG) and/or the reduced/oxidized thioredoxin ratio. Redox-sensitive green fluorescent proteins (GFP) allow real time visualization of the oxidation state of the indicator. Ratios of fluorescence from excitation at 400 and 490 nm indicate the extent of oxidation and thus the redox potential while canceling out the amount of indicator and the absolute optical sensitivity. Because the indicator is genetically encoded, it can be targeted to specific proteins or organelles of interest and expressed in a wide variety of cells and organisms. We evaluated roGFP1 (GFP with mutations C48S, S147C, and Q204C) and roGFP2 (the same plus S65T) with physiologically or toxicologically relevant oxidants both in vitro and in living mammalian cells. Furthermore, we investigated the response of the redox probes under physiological redox changes during superoxide bursts in macrophage cells, hyperoxic and hypoxic conditions, and in responses to H 2 O 2 -stimulating agents, e.g. epidermal growth factor and lysophosphatidic acid.Cells have elaborate homeostatic mechanisms to regulate the thiol-disulfide redox status of their internal compartments. Most thiol groups within the cytoplasm are normally reduced. Very few are present as disulfides. It has been speculated that the cytoplasm is reducing because many metabolic reactions evolved before oxygen became abundant in the atmosphere (1). Modest alterations in the thiol-disulfide equilibrium could have major consequences in the cell, including defective protein folding or enzyme activity (because many enzymes have a cysteine in their active site). When excess oxidation overwhelms the reductive capabilities of the cell, death results. Despite the dangers of excessive oxidation, cells sometimes use redox adjustments as signaling events, such as in the activation of transcription factors (NF-B and AP-1), caspases, protein tyrosine phosphatases, or GTPases (Ras). Thus, changes in the redox equilibrium influence a host of cell functions, including but not limited to growth, stress responses, differentiation, metabolism, cell cycle, communication, migration, gene transcription, ion channels, and immune responses (for reviews see Refs. 2-6). Alterations in the redox equilibrium are reflected in changes of the glutathione/glutathione-disulfide ratio (GSH/ GSSG) and the reduced/oxidized thioredoxin ratio. Glutathione is found in high concentrations in cells (5-10 mM) and is considered to be the major player in maintaining intracellular redox equilibrium. Ratios of GSH to GSSG are reported to range from 100 to 300:1 (7, 8), but these measurements have been problematic because they require destruction of the tissue, during which great care must be taken not to allow further oxidation. The major source of error is the determination of GSSG concentration, because this species is at low abundance yet is ...

show abstract

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Imaging Dynamic Redox Changes in Mammalian Cells with Green Fluorescent Protein Indicators

Dooley

Dore

Hanson

et al. 2004

Journal of Biological Chemistry

688

698

View full text Add to dashboard Cite

show abstract

“…In vivo, the coupled actions of diverse enzymes keep the concentration of this compound under the levels which may compromise the viability of a given organism [54,55]. However, under in vitro conditions the situation becomes quite different; hydrogen peroxide may become accumulated reaching high concentrations or it may require to be added to the reaction medium.…”

mentioning

confidence: 99%

Hydrogen Peroxide in Biocatalysis. A Dangerous Liaison

Hernández

Berenguer-Murcia²,

Rodrigues³

et al. 2012

COC

135

107

View full text Add to dashboard Cite

Hydrogen peroxide is a substrate or side-product in many enzyme-catalyzed reactions. For example, it is a side-product of oxidases, resulting from the re-oxidation of FAD with molecular oxygen, and it is a substrate for peroxidases and other enzymes. However, hydrogen peroxide is able to chemically modify the peptide core of the enzymes it interacts with, and also to produce the oxidation of some cofactors and prostetic groups (e.g., the hemo group). Thus, the development of strategies that may permit to increase the stability of enzymes in the presence of this deleterious reagent is an interesting target. This enhancement in enzyme stability has been attempted following almost all available strategies: site-directed mutagenesis (eliminating the most reactive moieties), medium engineering (using stabilizers), immobilization and chemical modification (trying to generate hydrophobic environments surrounding the enzyme, to confer higher rigidity to the protein or to generate oxidation-resistant groups), or the use of systems capable of decomposing hydrogen peroxide under very mild conditions. If hydrogen peroxide is just a side-product, its immediate removal has been reported to be the best solution. In some cases, when hydrogen peroxide is the substrate and its decomposition is not a sensible solution, researchers coupled one enzyme generating hydrogen peroxide "in situ" to the target enzyme resulting in a continuous supply of this reagent at low concentrations thus preventing enzyme inactivation.This review will focus on the general role of hydrogen peroxide in biocatalysis, the main mechanisms of enzyme inactivation produced by this reactive and the different strategies used to prevent enzyme inactivation caused by this "dangerous liaison". Outlook

show abstract

“…hydrogen peroxide, formed in vivo as a product of two general mechanisms: enzymatic or non-enzymatic dismutation of superoxide anions. H 2 O 2 is a physiological bacteriostatic agent, and a key element in the defence mechanisms of inflammatory cells; however, it is also involved in multiple physiological and pathological processes, including atherosclerosis, hypertension, mutagenesis, and cancer [40]. H 2 O 2 may alter protein conformation by oxidation of cysteine and methionine residues and, as a secondary messenger and modulator of gene expression, H 2 O 2 can influence signal transduction pathways, leading to the synthesis of cytokines and growth factors, such as interleukin 6 (IL-6) [41] or TGF-b [42].…”

Section: Disscusionmentioning

confidence: 99%

Red cabbage anthocyanins may protect blood plasma proteins and lipids

et al. 2011

View full text Add to dashboard Cite

The present in vitro study was designed to examine the antioxidative activity of red cabbage anthocyanins (ATH) in the protection of blood plasma proteins and lipids against damage induced by oxidative stress. Fresh leaves of red cabbage were extracted with a mixture of methanol/distilled water/0.01% HCl (MeOH/H2O/HCl, 50/50/1, v/v/w). Total ATH concentration [µM] was determined with cyanidin 3-glucoside as a standard. Phenolic profiles in the crude red cabbage extract were determined using the HPLC method. Plasma samples were exposed to 100 µM peroxynitrite (ONOO−) or 2 mM hydrogen peroxide (H2O2) in the presence/absence of ATH extract (5–15 µM); oxidative alterations were then assessed. Pre-incubation of plasma with ATH extract partly reduced oxidative stress in plasma proteins and lipids. Dose-dependent reduction of both ONOO− and H2O2-mediated plasma protein carbonylation was observed. ATH extract partly inhibited the nitrative action of ONOO−, and significantly decreased plasma lipid peroxidation caused by ONOO− or H2O2. Our results demonstrate that anthocyanins present in red cabbage have inhibitory effects on ONOO− and H2O2-induced oxidative stress in blood plasma components. We suggest that red cabbage ATH, as dietary antioxidants, should be considered as potentially usable nutraceuticals in the prevention of oxidative stress-related diseases.

show abstract

Role of hydrogen peroxide and oxidative stress in healing responses

Cited by 218 publications

References 227 publications

Imaging Dynamic Redox Changes in Mammalian Cells with Green Fluorescent Protein Indicators

Imaging Dynamic Redox Changes in Mammalian Cells with Green Fluorescent Protein Indicators

Hydrogen Peroxide in Biocatalysis. A Dangerous Liaison

Red cabbage anthocyanins may protect blood plasma proteins and lipids

Contact Info

Product

Resources

About