Damage of cell membrane and antioxidative system in human erythrocytes incubated with microcystin-LR in vitro.

The effect of the exposure of human erythrocytes to different concentrations of microcystin-LR were studied. Lipid peroxidation, membrane fluidity, cell morphology, haemoglobin oxidation and changes in the activity of antioxidant enzymes were investigated. Human erythrocytes were incubated with microcystin-LR at concentrations of 1-1000 nM for 1, 6, 12 and 24 h. We observed that microcystin-LR induces a significant increase of the level of thiobarbituric acid reactive substances (TBARS), formation of echinocytes, haemolysis, conversion of oxyhaemoglobin to methaemoglobin, decrease of membrane fluidity on the level of 16 carbon atom fat acids. The compound also changed antioxidative enzymes activities: catalase, superoxide dismutase and glutathione reductase and formation of reactive oxygen species (ROS). All of the observed changes point out that 100 nM of Microcistin LR is the liminal (threshold) toxic dose for human erythrocytes. This dose caused most of the described changes. Observed damages of erythrocytes membrane and antioxidative enzymes may be the result of direct covalent binding of microcystin-LR with -SH residues of proteins and indirectly be related with reactive oxygen species formation.     

Influence of dietary ginger (Zingiber officinales Rosc) on oxidative stress induced by malathion in rats.

Pesticide chemicals may induce oxidative stress leading to generation of free radicals and alterations in antioxidants or oxygen free radical (OFR) scavenging enzymes. Hence, the effect of subchronic malathion (O,O-dimethyl-S-1,2, bis ethoxy carbonyl ethyl phosphorodithioate) exposure was evaluated on lipid peroxidation, glutathione and related enzymes and OFR scavenging enzymes in albino rats. Administration of malathion (20 ppm) for 4 weeks increased the malondialdehyde (MDA) levels in serum, activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) in erythrocytes and glutathione reductase (GR) and glutathione S-transferase (GST) in serum. However, it decreased the glutathione (GSH) level in whole blood. Concomitant dietary feeding of Zingiber officinales Rosc (ginger 1%, w/w) significantly attenuated malathion induced lipid peroxidation and oxidative stress in these rats. These results indicate the possible involvement of free radicals in organophosphate-induced toxicity and highlight the protective action of ginger, an indigenous medicinal plant product.     

Alterations in human red blood cell properties induced by 3-(dimethylamino)phenol (in vitro)

3-(Dimethylamino)phenol (3-DMAP) exists in the environment as a transformation product of ureic herbicides and may also be considered as a derivative of phenoxyherbicides.

In this study, the activity of glutathione peroxidase, catalase and superoxide dismutase, as well as the level of free radicals and changes in cell morphology were measured in human erythrocytes exposed (in vitro) to 3-(dimethylamino)phenol. Human erythrocytes were incubated for 1 h in 3-DMAP at concentrations of 10–500 μg per 1 ml erythrocytes of 5% haematocrit.

The results show that 3-(dimethylamino)phenol increased the level of free radicals and changed the activity of glutathione peroxidase, catalase, superoxide dismutase and acetylcholinesterase. It also changed cell morphology.

All these results corroborated the thesis that 3-DMAP induces oxidative stress in cells. 3-DMAP changed the properties of the cell membrane, caused strong oxidation of haemoglobin, inhibited the levels of enzymatic and non-enzymatic antioxidants, which, in result, lead to generation of free radicals (ROS and semiquinones) that occurred in the exposed cells, predisposing them to oxidative damage.     

Effects of trichlorfon on malondialdehyde and antioxidant system in human erythrocytes

Organophosphorus insecticides may induce oxidative stress leading to generation of free radicals and alteration in antioxidant system. The aim of this study was to examine the potency of trichlorfon, an organophosphate insecticide, to induce oxidative stress response in human erythrocytes in vitro. For this purpose trichlorfon solutions in different concentrations and erythrocyte solutions were incubated at 37 °C for 60 min. At the end of the incubation time, malondialdehyde (MDA), an end product of lipid peroxidation, total glutathione, reduced glutathione (GSH) levels, activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) enzymes were determined by spectrophotometric methods. Trichlorfon increased MDA formation depended on the concentration. On the other hand, decreases in the GSH-Px activity, GSH levels and increases in the total glutathione levels, SOD and CAT activities were seen in the studied concentrations. The present findings indicate that the in vitro toxicity of trichlorfon may be associated with oxidative stress.     

Garlic-induced alteration in rat liver and kidney morphology and associated changes in endogenous antioxidant status.

The effects of chronic garlic intake on various endogenous antioxidant enzymes and lipid peroxidation on two major organs, the liver (L) and kidneys (K), were investigated. Wistar albino rats were fed with fresh garlic homogenate daily by gavage in three different doses (250, 500 and 1000 mg/kg/day) for 30 days. After this period, rats were sacrificed and liver and kidneys were harvested for biochemical estimation. In comparison to saline-treated rats, the 250 mg/kg/day dose significantly (P<0.02) reduced thiobarbituric acid reactive substances (TBARS) (L: 187.48+/-9.23 vs 150.66+/-11.45; K: 177.38 15.88 vs 120.66+/-9.39 nmol/g wet. weight) and glutathione peroxidase (GPx) (L: 0.2438+/-0.05 vs 0.0046+/-0.0005; K: 0.1459+/-0.034 vs 0.0055+/-0.0003 U/mg protein). There was no change in catalase and reduced glutathione (GSH) but superoxide dismutase (SOD) increased significantly (P<0.01) (L: 5.49+/-0.76 vs 18.38+/-2.26; K: 11.47+/-1.48 vs 21.22+/-3.19 U/mg protein). Both 500 and 1000 mg/kg/day doses significantly (P<0.05) reduced endogenous antioxidants (catalase and SOD) without altering TBARS. A 1000 mg/kg/day dose of garlic caused marked histopathological and ultrastructural changes in both liver and kidneys. The results suggest that garlic in low doses has the potential to enhance the endogenous antioxidant status, although at higher doses a reversal of these effects is observed. The present study also highlights the potential ability of a high dose of garlic to induce morphological changes in the liver and kidneys, indicating the need to identify a safe dose range for garlic.     

Counteraction by glutathione and selenium of the prooxidant effect of alloxan in erythrocytes in vitro

Alloxan produces reactive oxygen species which cause injury to the insulin-producing β-cells of the pancreas. The action of alloxan in the production of free radicals was studied in vitro using rat erythrocytes. Alloxan increased lipid peroxidation (LPO) and decreased the activities of the antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase in the erythrocytes. Treatment with glutathione (GSH) and/or selenium along with alloxan was able to decrease LPO and increase the activities of the antioxidant enzymes in the erythrocytes. The results suggest that GSH and selenium both counteract the prooxidant effect of alloxan in rat erythrocytes.     

In vitro effect of n-nitrosodiethylamine on lipid peroxidation and antioxidant system in human erythrocytes

Human erythrocytes were used in vitro to investigate the effect of the hepatocarcinogen N-nitrosodiethylamine (NDEA) on lipid peroxidation (LPO) and antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR). LPO and erythrocyte haemolysis increased with increasing concentrations of NDEA and with increased exposure time. CAT activity decreased while GR activity increased with both the increasing concentrations of NDEA treatment and exposure time. However, no alteration was observed in SOD enzyme activity. The inhibitory effects of antioxidants and free radical scavengers such as EDTA, succinic acid, sodium benzoate and ascorbic acid were observed. These agents lowered NDEA-induced LPO and haemolysis in erythrocytes. This might indicate that the generation of free radicals and subsequent LPO may play a role, at least in part, in inducing NDEA toxicity.     

Biochemical evidence for free radical-induced lipid peroxidation as a mechanism for subchronic toxicity of malathion in blood and liver of rats

Organophosphorus compounds may induce oxidative stress leading to generation of free radicals and alterations in antioxidant and scavengers of oxygen free radicals (OFRs). The effect of subchronic exposure to malathion in the production of oxidative stress was evaluated in male Wistar rats. Administration of malathion (100, 316, 1000, 1500 ppm) for 4 weeks increased catalase (CAT), superoxide dismutase (SOD) activities as well as malondialdehyde (MDA) concentration in red blood cells (RBC) and liver. However, acetylcholinesterase (AChE) and cholinesterase (ChE) activities were decreased in these samples. The increase in RBC and liver lipid peroxidation correlated well with the inhibition in RBC AChE and liver ChE activities. Elevation of MDA concentrations and increased activities of CAT and SOD showed significant correlations in both RBC and liver samples when different doses of malathion were used. The results of the present study suggest the usefulness of RBC AChE measurement as a good biomarker in the estimation of malathion-induced oxidative stress affecting blood and liver.     

Effects of smoking on plasma and erythrocyte antioxidant defense systems

In this study, activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) enzymes were measured in the erythrocytes, and levels of thiobarbituric acid-reactive substances (TBARS) and antioxidant potential (AOP) values were measured in both erythrocyte and plasma samples from smokerS and nonsmokers. No significant differences were observed in erythrocyte parameters, serum triglycerides, and total cholesterol. AOP was significantly lower and TBARS level higher in the plasma samples from smokers compared with those of nonsmokers. Results suggest that smoking causes no impairment in the enzymatic antioxidant defense system and does not lead to oxidant stress in the erythrocytes, possibly because these cells have potent antioxidant defense capacity.     

Studies on the mechanisms of oxidation in the erythrocyte by metabolites of primaquine

The interaction of certain metabolites of the 8-aminoquinoline antimalarial primaquine with both normal and glucose-6-phosphate dehydrogenase (G6PD)-deficient erythrocytes and with haemoglobin preparations was studied in an attempt to elucidate the mechanisms of methaemoglobin formation and haemolytic anaemia associated with the use of primaquine. Studies using erythrocytes revealed that oxidation of haemoglobin and reduced glutathione (GSH) was due to the metabolites rather than the parent drug. Incubation of free haemoglobin with 5-hydroxylated metabolites of primaquine also led to oxidation of oxyhaemoglobin and GSH. Oxidation of GSH also occurred in the absence of oxyhaemoglobin. The results suggest a dual mechanism for these oxidative effects, involving autoxidation of the 5-hydroxy-8-aminoquinolines and their coupled oxidation with oxyhaemoglobin. The initial products of these processes would be drug metabolite free radicals, superoxide radical anions, hydrogen peroxide and methaemoglobin. Further free radical reactions would lead to oxidation of GSH, more haemoglobin and probably other cellular constituents. NADPH had no effect on the oxidative effects of the primaquine metabolites in these experiments. In the G6PD-deficient erythrocyte, the oxidation of haemoglobin and GSH leads to Heinz body formation and eventually to haemolysis, the mechanisms of which are as yet unclear. The possible role of oxygen free radicals in the mode of action of 8-aminoquinolines against the malaria parasite is also briefly discussed.     

Inhibitory effects of various sulfur compounds on the activity of bovine erythrocyte enzymes

Studies were conducted to assess the in vitro effects of selected sulfur compounds on the activities of superoxide dismutase (SOD), catalase, glutathione peroxidase (GSHPX), and glucose-6-phosphate dehydrogenase (G6PDH) in hemolyzates of bovine erythrocytes. All sulfur compounds produced concentration-dependent inhibition in the activities of these enzymes, but their effects on each enzyme were different. SOD and catalase activities were most sensitive to sulfide (S2-), followed by sulfite (SO3(2-)) and sulfate (SO4(2-)). GSHPX activity was most sensitive to SO3(2-), followed by S2-, cysteine and SO4(2-). The activity of G6PDH, however, was maximally inhibited by reduced glutathione (GSH), followed by SO3(2-) and SO4(2-); S2- was inhibitory only at high concentrations. Dialysis of the S2- and SO3(2-)-inhibited enzymes resulted in complete or partial reversal of inhibitory effects. The biochemical significance of these effects in relation to erythrocyte physiology is discussed.     

Effect of fluoride intoxication on lipidperoxidation and antioxidant status in experimental rats

Fluoride is a potent enzyme poison. Thirty ground water samples from Vellore District, Tamil Nadu, India were analysed for fluoride content and it was revealed that the fluoride content of 24 samples were over and above the permissible limits. In the present study, the experimental rats were orally treated with 25 ppm of fluoride/rat/day for 8 and 16 weeks, respectively, and the levels of lipid peroxidation and antioxidant enzymes were studied to evaluate fluoride intoxication. An increase in the level of lipid peroxides along with a concomitant decrease in the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and reduced glutathione content were observed in fluoride administered groups of rats. The altered antioxidant status may be attributed to the increased generation of free radicals.     

Gene expression and activity of antioxidant enzymes in the blood cells of workers who were occupationally exposed to lead

In this study, we sought to understand the influence of occupational lead-exposure on the gene expression (Sod1) and activity (SOD) of superoxide dismutase, catalase and glutathione peroxidase (GPx, Gpx1) in leukocytes and erythrocytes. The study group consisted of 45 healthy male employees of a lead-zinc works and was divided into two subgroups: those with low exposure to lead (LE) and those with high exposure to lead (HE). In addition, 17 healthy male administrative workers participated in the study as the control group. The gene expression levels of both Sod1 and Gpx1 were significantly increased in the LE group as compared to the control group. By contrast, we noted only an insignificant tendency for increased gene expression of both Sod1 and Gpx1 in the HE group. The expression and activity of catalase were unchanged. Nevertheless, SOD and GPx activities in erythrocytes was significantly elevated in both examined subgroups, whereas SOD activity in leukocytes was raised only in the LE group. The results of this study led us to conclude that lead has a significant influence not only on the activities of antioxidant enzymes but also on the dose-dependent expression in their genes.     

Effects of deltamethrin on lipid peroxidation in mice

Effects of deltamethrin on lipid peroxidation were investigated for subacute, subchronic and chronic periods. Deltamethrin was given to Swiss Albino mice at 1.5, 2.5, or 7.5 mg/kg. Lipid peroxidation was evaluated by determining malondialdehyde (MDA) levels in plasma, and determining glutathione peroxidase (GSH-Px), superoxide dismutase (Cu-Zn SOD) and catalase activities in erythrocytes on days 15, 45, and 60. MDA levels increased in deltamethrin dosed groups, especially for the subchronic and chronic periods. GSH-Px, Cu-Zn SOD and catalase activities in erythrocytes were decreased at high doses of deltamethrin, especially at the high dose (7.5 mg/kg) deltamethrin group.     

Myricetin suppresses oxidative stress-induced cell damage via both direct and indirect antioxidant action

We evaluated the cytoprotective effect of myricetin on oxidative stress damaged cells by assessment of the scavenging effect of reactive oxygen species (ROS) and the activities of antioxidant enzymes. Myricetin showed the scavenging effect of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals on intracellular ROS. In addition, myricetin restored the activity and protein expression of cellular antioxidant defense enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) reduced by hydrogen peroxide (H(2)O(2)) treatment. H(2)O(2)-induced cellular DNA and lipid damages, and myricetin was found to prevent the DNA damage shown by inhibition of DNA tail and it decreased nuclear phospho-histone H2A.X expression, which are both markers for DNA strand breakage. Membrane lipid peroxidation was also attenuated as shown by inhibition of TBARS formation and of fluorescence intensity of diphenyl-1-pyrenylphosphine (DPPP). These results suggest that myricetin protects cells against H(2)O(2)-induced cell damage via inhibition of ROS generation and activation of antioxidant enzymes.     

Studies with primaquine in vitro: superoxide radical formation and oxidation of haemoglobin.

1. The production of superoxide radicals from primaquine diphosphate in aqueous solution has been demonstrated, using as indicator the reduction of cytochrome C with inhibition of the reaction by superoxide dismutase. 2. Primaquine-mediated oxidation of haemoglobin to methaemoglobin was reduced by the addition of catalase and increased by superoxide dismutase. Mannitol, a hydroxyl radical scavenger, abolished the increase in methaemoglobin observed in the presence of superoxide dismutase. EDTA reduced the oxidation of haemoglobin with and without superoxide dismutase. 3. Although the oxidation of haemoglobin in the presence of primaquine includes the effects of hydrogen peroxide, superoxide and hydroxyl radicals and metal ions, the results indicate that hydrogen peroxide, rather than the superoxide radical, is the main oxidizing species. The increase in haemoglobin oxidation occurring with superoxide dismutase may result from the augmented rate of hydrogen peroxide formation from superoxide radicals.     

Evaluation of protective effect of rutin on lead acetate-induced testicular toxicity in Wistar rats

In this study, the effect of rutin (RT) was investigated on lead-induced testis tissue damage in rats. Oral administration of rutin (50?mg/kg) effectively inhibited the levels of marker enzymes and antioxidant enzymes as compared with lead acetate-treated group. Antioxidant enzyme activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST), and histopathological changes of testes were investigated. Lead acetate decreased the levels of SOD, CAT, GPx, and GST compared with the control group. Light microscopic analyses revealed that lead acetate induced several histopathological changes in testis tissue. In the RT-treated group, there were statistically significantly decreases in antioxidant enzyme activities and pathological changes in the tissue. The results suggest that RT possesses significant potential in reduced lead acetate-induced testicular toxicity.     

Gastroprotective and antioxidant effects of amiodarone on indomethacin-induced gastric ulcers in rats

Reactive oxygen species (ROS) have been implicated in the etiology of indomethacin-induced gastric mucosal damage. This study investigated amiodarone's protective effects against oxidative gastric mucosal damage induced by indomethacin. Amiodarone is a widely used antiarrhythmic agent. We have investigated alterations in the glutathione level, and the activities of antioxidative enzymes [superoxide dismutase, catalase, glutathione s-transferase glutathione reductase and myeloperoxidase], as markers for ulceration process following oral administration of amiodarone and ranitidine in rats with indomethacin-induced ulcers. In the present study we found that 1) amiodarone, lansoprazole and ranitidine reduced the development of indomethacin-induced gastric damages, at a greater magnitude for amiodarone and lansoprazole than for ranitidine; 2) amiodarone and ranitidine alleviated increases in the activities of catalase and glutathione s-transferase enzymes resulting from ulcers; 3) amiodarone and ranitidine ameliorated depressions in the glutathione level and the activities of superoxide dismutase and glutathione reductase enzymes caused by indomethacin administration; and 4) all doses of amiodarone amplified the myeloperoxidase activity resulting from indomethacin-induced gastric ulcers. The results indicate that the gastroprotective activity of amiodarone, which may be linked to its intrinsic antioxidant properties, cannot be attributed to its effect on myeloperoxidase activity.     

Advances in metal-induced oxidative stress and human disease

Detailed studies in the past two decades have shown that redox active metals like iron (Fe), copper (Cu), chromium (Cr), cobalt (Co) and other metals undergo redox cycling reactions and possess the ability to produce reactive radicals such as superoxide anion radical and nitric oxide in biological systems. Disruption of metal ion homeostasis may lead to oxidative stress, a state where increased formation of reactive oxygen species (ROS) overwhelms body antioxidant protection and subsequently induces DNA damage, lipid peroxidation, protein modification and other effects, all symptomatic for numerous diseases, involving cancer, cardiovascular disease, diabetes, atherosclerosis, neurological disorders (Alzheimer's disease, Parkinson's disease), chronic inflammation and others. The underlying mechanism of action for all these metals involves formation of the superoxide radical, hydroxyl radical (mainly via Fenton reaction) and other ROS, finally producing mutagenic and carcinogenic malondialdehyde (MDA), 4-hydroxynonenal (HNE) and other exocyclic DNA adducts. On the other hand, the redox inactive metals, such as cadmium (Cd), arsenic (As) and lead (Pb) show their toxic effects via bonding to sulphydryl groups of proteins and depletion of glutathione. Interestingly, for arsenic an alternative mechanism of action based on the formation of hydrogen peroxide under physiological conditions has been proposed. A special position among metals is occupied by the redox inert metal zinc (Zn). Zn is an essential component of numerous proteins involved in the defense against oxidative stress. It has been shown, that depletion of Zn may enhance DNA damage via impairments of DNA repair mechanisms. In addition, Zn has an impact on the immune system and possesses neuroprotective properties. The mechanism of metal-induced formation of free radicals is tightly influenced by the action of cellular antioxidants. Many low-molecular weight antioxidants (ascorbic acid (vitamin C), alpha-tocopherol (vitamin E), glutathione (GSH), carotenoids, flavonoids, and other antioxidants) are capable of chelating metal ions reducing thus their catalytic activity to form ROS. A novel therapeutic approach to suppress oxidative stress is based on the development of dual function antioxidants comprising not only chelating, but also scavenging components. Parodoxically, two major antioxidant enzymes, superoxide dismutase (SOD) and catalase contain as an integral part of their active sites metal ions to battle against toxic effects of metal-induced free radicals. The aim of this review is to provide an overview of redox and non-redox metal-induced formation of free radicals and the role of oxidative stress in toxic action of metals.     

Effect of Herbal Preparation on Heavy Metal (Cadmium) Induced Antioxidant System in Female Wistar Rats

Cadmium is one of the elements found to damage antioxidant systems in mammals. To ameliorate cadmium toxicity and to prevent oxidative stress, natural products may be useful. In Indian ethnobotanical practice, a mixture of 17 herbal products is used to fortify the reproductive system of women after parturition and to reverse ovarian oxidative stress. Oral administration of this extract to rats exposed to cadmium was useful in reversing oxidative stress. Two different doses of cadmium (50 ppm and 200 ppm) were given to Wistar rats aged 45 and 65 days. An herbal extract derived from 17 plants was administered orally every day at a dose level of 200 mg/kg of body weight to the rats exposed to cadmium. A battery of enzymes involved in antioxidant activity in the ovary, including superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx) and glutathione-s-transferase (GST) were measured in the control, cadmium-exposed rats without treatment and in the cadmium-exposed rats treated with herbal extract. The reduction in SOD, catalase, GPx and GST activity after cadmium exposure improved significantly in the rats treated with the herbal extract (p?相似文献