Grzegorz Bartosz

Reactive oxygen species: Destroyers or messengers?

Abundant evidence leaves no doubt that reactive oxygen species (ROS) are not only inevitable by-products of oxygen metabolism but also play a role in cellular signaling. ROS are produced by a family of NADPH oxidases for signaling purposes and mediate or augment the effects of insulin, growth factors, cytokines and G-protein-coupled receptors. Disturbances of ROS signaling leading to overproduction of these intermediates inflict oxidative damage of cell components in the course of various diseases. Restoration of proper ROS signaling, especially inhibition of cellular sources of ROS, may thus provide new ways of therapy.

Antioxidative and prooxidative effects of quercetin on A549 cells

Quercetin, a common plant polyphenol, has been reported to show both antioxidant and prooxidant properties. We studied the effects of quercetin on A549 cells in in vitro culture. We found that low concentrations of the flavonoid stimulated cell proliferation and increased total antioxidant capacity (TAC) of the cells; while higher concentrations of the flavonoid decreased cell survival and viability, thiol content, TAC and activities of superoxide dismutase, catalase and glutathione S‐transferase. Quercetin decreased production of reactive oxygen species in the cells but produced peroxides in the medium. The cellular effects of quercetin are therefore complex and include both antioxidant effects and induction of oxidative stress due to formation of reactive oxygen species in the extracellular medium.

Apoptosis-like, reversible changes in plasma membrane asymmetry and permeability, and transient modifications in mitochondrial membrane potential induced by curcumin in rat thymocytes

Curcumin (diferuoylmethane) is a natural compound with anticarcinogenic activities which is able to exert either proapoptotic or antiapoptotic effects in different cell types. This paper focuses on the sequence and extent of primary events induced by curcumin, in comparison with those occurring during dexamethasone‐induced apoptosis in rat thymocytes. It also presents annexin VI‐FITC as a new probe for studying membrane asymmetry. Curcumin readily penetrates into the cytoplasm, and is able to accumulate in membranous structures such as plasma membrane, endoplasmic reticulum and nuclear envelope. Curcumin‐treated cells exhibit typical features of apoptotic cell death, including shrinkage, transient phosphatidylserine exposure, increased membrane permeability and decrease in mitochondrial membrane potential. However, nuclei morphology, DNA fragmentation, the extent and time‐course of membrane changes are different from those observed during dexamethasone‐induced apoptosis, suggesting that, despite many similarities, the mode of action and the events triggered by curcumin are different from those occurring during typical apoptosis.

Effect of Vitamin E and Vitamin C Supplementation on Antioxidative State and Renal Glomerular Basement Membrane Thickness in Diabetic Kidney

The aim of this study was to analyze the effect of vitamins C and E on malondialdehyde (MDA) content and activities of key antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) as well as glomerular basement membrane (GBM) thickness in streptozotocin-induced diabetic kidney in rats. Wistar male rats were divided into following groups (12 rats each): the control, diabetic rats, diabetic rats whose drinking water was supplemented with vitamin C in a dose of 1.0 g/l or diet was supplemented with 200 mg of vitamin E/100 g fodder. Body weight, blood glucose and HbA1C levels and 24-hour urinary albumin excretion (UAE) were studied every week (0–12 weeks). After 6 and 12 weeks, MDA content and activities of SOD, CAT and GSH-Px were measured in the kidney homogenate supernatants. Electron micrographs of glomeruli were scanned and morphometric investigations were performed by means of computer image analysis system to compare GBM thickness. The blood glucose and HbA1C concentrations and UAE in diabetic rats were significantly higher than in the control group. An increase in the MDA level and decrease in the SOD, CAT and GSH-Px activities in the kidney of diabetic rats were observed after 6 and 12 weeks of experiment. Administration of vitamins C and E did not affect body weight, blood glucose and HbA1C levels. Both vitamin C and vitamin E decreased lipid peroxidation and augmented the activities of antioxidant enzymes studied in the kidneys of diabetic rats as well as reduced UAE, decreased kidney weight and GBM thickness. The results indicate the potential utility of antioxidant vitamins in the protection against the development of diabetic nephropathy.

Natural and synthetic antioxidants: An updated overview

Abstract The current understanding of the complex role of ROS in the organism and pathological sequelae of oxidative stress points to the necessity of comprehensive studies of antioxidant reactivities and interactions with cellular constituents. Studies of antioxidants performed within the COST B-35 action has concerned the search for new natural antioxidants, synthesis of new antioxidant compounds and evaluation and elucidation of mechanisms of action of both natural and synthetic antioxidants. Representative studies presented in the review concern antioxidant properties of various kinds of tea, the search for new antioxidants of herbal origin, modification of tocopherols and their use in combination with selenium and properties of two promising groups of synthetic antioxidants: derivatives of stobadine and derivatives of 1,4-dihydropyridine.

Non-enzymatic antioxidant capacity assays: Limitations of use in biomedicine.

Abstract The ‘Total antioxidant capacity’ (TAC) is a parameter frequently used for characterization of food products and of the antioxidant status of the body. This mini-review shows shortcomings of TAC assays and points of concern that should be considered when performing and interpreting results of such assays. The term TAC is not optimal since the assay measures only part of antioxidant capacity, usually excluding enzymatic activities. Antioxidant and oxidant-regenerating enzymes in blood cells and the blood vessel wall have a profound impact on the antioxidant properties of blood plasma, which is not reflected in the in vitro assays of isolated plasma. The term ‘Non-enzymatic antioxidant capacity’ (NEAC) is suggested as more relevant than TAC. NEAC is estimated by various methods, which yield different values and results obtained using different methods do not always show satisfactory correlation. One reason for the discrepancy of results is the use of different oxidants in NEAC assays. The use of hydroxyl radical as the oxidant is not recommended in view of the high and non-specific reactivity of this species.

DECREASE IN ACCESSIBLE THIOLS AS AN INDEX OF OXIDATIVE DAMAGE TO MEMBRANE PROTEINS

The effect of several oxidative agents (hydrogen peroxide, tert-butyl hydroperoxide, menadione, AAPH, peroxynitrite and ionizing radiation) on the ratio of weakly to strongly immobilized residues of erythrocyte membrane-bound maleimide-tempo spin label (h(w)/h(s) ratio) was studied in order to test the hypothesis that a decrease in the h(w)/h(s) ratio may be a general index of oxidative damage to membrane proteins. Most of the agents studied decreased though H2O2 almost did not affect and ionizing radiation increased the h(w)/h(s) ratio. In parallel, the ratio of DTNB accessible/DTNB inaccessible membrane protein-SH groups was determined from membrane-SH group measurements with the Ellman reagent in the absence and in the presence of sodium dodecyl sulfate. This ratio decreased in all cases studied and seems to be a more universal and easy to measure parameter to describe the oxidative damage to membrane proteins.

Effect of Antioxidants Supplementation on Aging and Longevity

If aging is due to or contributed by free radical reactions, as postulated by the free radical theory of aging, lifespan of organisms should be extended by administration of exogenous antioxidants. This paper reviews data on model organisms concerning the effects of exogenous antioxidants (antioxidant vitamins, lipoic acid, coenzyme Q, melatonin, resveratrol, curcumin, other polyphenols, and synthetic antioxidants including antioxidant nanoparticles) on the lifespan of model organisms. Mechanisms of effects of antioxidants, often due to indirect antioxidant action or to action not related to the antioxidant properties of the compounds administered, are discussed. The legitimacy of antioxidant supplementation in human is considered.

Effect of peroxynitrite on erythrocytes

The action of peroxynitrite on human erythrocytes and erythrocyte membranes was studied. Peroxynitrite (0.1-2 mM) induced a transient decrease of intracellular reduced glutathione, oxidized membrane protein -SH groups, initiated membrane lipid peroxidation and inactivated erythrocyte membrane acetylcholinesterase and ATPase activities. Membranes exposed to peroxynitrite showed aggregation and nitration of proteins and changes in protein organization detectable with a maleimide spin label.

Molecular strategies to prevent, inhibit, and degrade advanced glycoxidation and advanced lipoxidation end products

Abstract The advanced glycoxidation end products (AGEs) and lipoxidation end products (ALEs) contribute to the development of diabetic complications and of other pathologies. The review discusses the possibilities of counteracting the formation and stimulating the degradation of these species by pharmaceuticals and natural compounds. The review discusses inhibitors of ALE and AGE formation, cross-link breakers, ALE/AGE elimination by enzymes and proteolytic systems, receptors for advanced glycation end products (RAGEs) and blockade of the ligand–RAGE axis.