Erythrocyte Membrane Damage Research Articles

The effects of the caffeic acid phenethyl ester (CAPE) on erythrocyte membrane damage after hind limb ischaemia-reperfusion.

Reactive oxygen species have been implicated in pathogenesis injury after ischaemia-reperfusion (I/R). Caffeic Acid Phenethyl Ester (CAPE), an active component of honeybee propolis extract, exhibits antioxidant and anti-inflammatory properties. The aim of this study was to investigate the effects of CAPE on erythrocyte membrane damage after hind limb I/R. Rats were divided into two groups: I/R and I/R with CAPE pre-treatment. They were anaesthetized with intramuscular ketamine 100 mg kg(-1). A 4-h I/R period was performed on the right hind limb of all animals. In the CAPE-treated group, animals received CAPE 10 microm by intraperitoneal injection 1 h before the reperfusion. At the end of the reperfusion period, a midsternotomy was performed. A 5-ml blood sample was withdrawn from the ascending aorta for biochemical assays. Serum and erythrocyte membrane MDA levels were significantly lower in the CAPE-treated group when compared to the I/R group ( p = 0.001 and p<0.001, respectively). Erythrocyte membrane Na(+)-K(+) ATPases activity in the CAPE-treated group was significantly higher than the I/R group ( p<0.001). In conclusion, CAPE seems to be effective in protecting against oxidative stress. Therefore, we suggest that in order to decrease I/R injury, pre-administration of CAPE may be a promising agent for a variety of conditions associated with I/R.

Alterations in Skeletal Protein, Distribution of PKCα, and Level of Phospholipids in Erythrocyte Membranes of Women with Primary Breast Cancer

ABSTRACT The aim of our work was to study the influence of primary breast cancer on mature erythrocyte membranes. Blood was sampled from 29 women with primary breast cancer, aged 35–86 years, in different stages of clinical progression of the disease. In red blood cell membranes an increase of phosphatidylinositol-4-phosphate and phosphatidylinositol-4,5-diphosphate levels was observed. These changes were accompanied by a decrease in phospholipase C activity. Simultaneously, a significant decrease in concentration of phosphatidylserine, sphingomyelin, and phosphatidylinositol was found. Quantitative protein evaluation showed an increase in band 4.1 protein content with no changes in the level of constitutive PKCα responsible for the phosphorylation of this protein and its affinity to glycophorine C. In parallel a greater increase of PKCα translocation after PMA treatment compared to controls was observed. Possible oxidative damage of erythrocyte membranes indicated by an increase in malonyldialdehyde level and decrease in SH-group content as well as by an increase in the w/ ratio was documented. From the results it is concluded that primary breast cancer seems to affect the membranes of mature erythrocytes.

Erythrocyte damage and leukocyte activation in ischemic stroke

Background: The traditional lipid risk factors can only predict some of the cardiovascular events. Our work has focused on new potential biological markers of risk, namely leukocyte activation and erythrocyte membrane damage, in ischemic stroke cases. Methods: Besides the traditional lipid profile, we evaluated the plasma levels of elastase and lactoferrin as markers of leukocyte activation, and membrane band 3 protein profile and membrane bound hemoglobin as markers of erythrocyte damage. Total and differential leukocyte counts and erythrocyte counts, hematocrit and hemoglobin concentrations were also evaluated. The lipid study included the evaluation of triglycerides, total cholesterol, high-density lipoprotein cholesterol (HDLc), low-density lipoprotein cholesterol (LDLc), apolipoprotein AI (Apo AI) and B (Apo B), and lipoprotein (a) (Lp(a)). The work was performed in a control group ( n=29) with no history of cardiovascular events, presenting normal hematological and lipid values, and in a pathologic group ( n=21) of ischemic stroke cases diagnosed by computed tomographic imaging. Results: We found that ischemic stroke was associated with significantly higher values of leukocytes, which seem to be activated, as shown by significant higher levels of elastase and lactoferrin. This activation seems to impose erythrocyte damage, as suggested by a significant increase in membrane bound hemoglobin and by a different band 3 profile. Conclusions: Our data suggest that plasma levels of elastase and lactoferrin, together with levels of erythrocyte membrane bound hemoglobin and band 3 profile, could be used as powerful new markers of risk for cardiovascular events.

Protection by taurine and structurally related sulfur-containing compounds against erythrocyte membrane damage by hydrogen peroxide.

Protection by taurine and structurally related sulfur-containing compounds against erythrocyte membrane damage by hydrogen peroxide.

Preventive effect of selenium on T-2 toxin membrane toxicity.

T-2 toxin, one of the major toxic trichothecene mycotoxines, has been shown to cause effects such as inhibition of protein synthesis and impairement of mitochondrial function. The use of T-2 toxin as chemical warfare in south east Asia and Iran has been reported . It has been suggested that T-2 toxin may mediate its toxic effect via the cell membrane, but mechanism of action is poorly understood. In cytotoxicity studies, erythrocytes are an excellent model system. In the present study different doses of sodium selenite were injected into male albino mice for 6 days every 48 h. Blood samples were taken from experimental and control groups (normal saline). The red cells were counted in isotonic phosphate buffer containing different doses of T-2 toxin. The mixture was incubated at 37 degrees C for 4 h. The results indicate that selenium is able to prevent erythrocyte membrane damage induced by T-2 toxin. The protective effect of selenium may be due to its membrane stabilizing properties, although inhibition of lipid peroxidation is likely, too.

Impaired erythrocyte transmembrane potential in diabetes mellitus and its possible improvement by resorcylidene aminoguanidine

Erythrocytes of diabetic patients have abnormal membrane properties. We examined in vitro transmembrane potential and the possible effect of resorcylidene aminoguanidine (RAG) on its modulation in erythrocytes of diabetic subjects. The transmembrane potential was assessed in RAG-treated and untreated erythrocytes, respectively, using a fluorescent dye (3,3′-dipropylthiadicarbocyanine iodide [DiSC 3(5)]). We confirmed earlier findings that the transmembrane potential of diabetic erythrocytes is significantly increased compared with control ( P<0.01). The membrane hyperpolarization found in diabetic cells seems to be a result of oxidative stress present in diabetes mellitus. On one hand, the RAG treatment induced decrease in abnormal transmembrane potential values in diabetic erythrocytes ( P<0.01), presumably via its antioxidant and antiglycation activity. On the other hand, RAG moderately hyperpolarized the control erythrocytes ( P<0.05). We suggest that the drug-induced transient membrane expansion leads to an intracellular potassium loss and a subsequent change of the transmembrane potential. However, if controlled by an appropriate dosage, RAG can eliminate certain types of erythrocyte membrane damage induced by diabetes mellitus.

Dye sorption as an indicator of erythrocyte membrane damage and prehemolytic state of erythrocytes

High erythrocyte membrane permeability caused by detergents was associated with increased methylene blue sorption, which correlated with erythrocyte damage and preceded hemolysis. The content of sorbed dye in erythrocytes preincubated with 200 μM sodium dodecyl sulfate or cetyltrimethylammonium bromide increased by 58–74% and 2.4–3.8-fold surpassed that in the medium. New precise and convenient method for estimating erythrocyte binding capacity is proposed.

Protective effect of erythropoietin on the oxidative damage of erythrocyte membrane by hydroxyl radical

Treatment of red blood cells with the copper (II) ascorbate system causes increased lipid peroxidation, increased membrane microviscosity, and phospholipid translocation with a concurrent decrease in cytosolic catalase and glutathione peroxidase activities. All these changes are prevented if the cells are treated with erythropoietin prior to the exposure to copper (II) ascorbate. The present investigation further indicates that the oxidative damage brought about by copper (II) ascorbate is due to generation of hydroxyl radical and that erythropoietin plays a unique role in protecting the membrane from oxidative damage.

Inhibition of iron ion-induced oxidative damage of erythrocyte membranes and low density lipoprotein by a Maillard product, 4-hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF).

4-Hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF) is representative of the Maillard reaction-derived reductones found in many foodstuffs. Influence of HEMF on iron ion-induced oxidative modification of human erythrocyte membranes and low density lipoprotein (LDL) under aerobic conditions was investigated. When human erythrocytes were incubated at 37 degrees C for 24 h with HEMF alone, levels of thiobarbituric acid-reactive substances (TBARS) and non-ionic detergent C12E8-insoluble membrane protein aggregates in the membranes were unchanged. Levels of TBARS and protein aggregates in the membranes increased on treatment of the cells with Fe(III) ion at 37 degrees C for 3 h were lowered in the presence of HEMF. Western blot analysis indicated that aggregates of band 3 protein induced by Fe (III) ion were decreased in the presence of HEMF, and the level of TBARS in LDL increased on treatment with Fe(II) or Fe(II) ion at 37 degrees C for 24 h was also lowered in the presence of HEMF. The results indicate that HEMF protected the iron ion-induced oxidative modification of human erythrocyte membranes and LDL.

Physicochemical properties and hemolytic effect of different lipid emulsion formulations using a mixture of emulsifiers

Abstract The purpose of this study was to elucidate the influence of ternary mixtures of different emulsifiers on the physicochemical properties of lipid emulsions by the aid of simplex lattice design with constraints. The physicochemical properties and the stability during the autoclaving process, as a consequent result, were investigated by examining the changes in particle size, zeta potential, and cloud point. The emulsifiers chosen were lecithin, Synperonic F68 and Tween 80. In addition, their potential for hemolysis was evaluated as it may serve as an in vitro toxicity screening method. The formulations with only Tween 80 as an emulsifier showed a noticeable change in the particle size during autoclaving as well as a remarkable erythrocyte membrane damage. In contrast, phospholipids and Synperonic F68 formulations displayed good stability during autoclaving and showed almost no hemolytic activity. Moreover, mixing Tween 80 with either lecithin or Synperonic F68 improved the stability of these formulations during the autoclaving process. Simultaneously, this led to a remarkable decrease in the hemolytic effect. This observation could be partially correlated with the cloud point, which increased by adding lecithin and was less influenced by adding Synperonic F68. This in turn resulted in an increase in the surface charge, when lecithin was added. This results in a higher resistance of the emulsifier complex to dehydration producing a more stable emulsifier film around the oil droplets. These factors inhibited the flocculation of the emulsifiers and could hinder the coalescence of the oil droplets during the autoclaving process.

Oxidative damage of erythrocyte membrane in nephrotic syndrome.

Erythrocytes are target cells for peroxidative damage. Abnormal susceptibility of erythrocyte lipids to peroxidation is believed to reflect a similar abnormality in other organs and tissues. The changes in erythrocyte lipid peroxidation [measured by malonyldialdehyde (MDA) concentration] and erythrocyte membrane cholesterol (EMC) and their correlation with plasma lipid changes were studied in 36 children with steroid-responsive minimal change nephrotic syndrome (MCNS) (16 in relapse, 20 in remission) and 30 matched healthy controls. Erythrocyte MDA levels were significantly higher in relapse [126.3+/-40.6 nmol/g hemoglobin (Hb)] compared with remission (101.2+/-21.3 nmol/g Hb, P<0.02) and in controls (95.4+/-20.4 nmol/g Hb, P<0.001). Plasma MDA levels in relapse were also higher than in remission (4.26+/-1.19 nmol/ml vs. 3.16+/-1.18 nmol/ml, P<0.01), and in controls (2.49+/-0.86 nmol/ml, P<0.001). The EMC content changed significantly during remission (1.22+/-0.15 mg/10(10) cells in relapse, 1.09+/-0.19 mg/10(10) cells in remission, P<0.04). These results show an increased sensitivity of red cells to lipid peroxidation in patients with steroid-sensitive nephrotic syndrome without the development of renal failure and anemia. Lipid peroxidation of plasma lipids and erythrocyte membrane may be a primary phenomenon, but this should be confirmed by investigation of peroxidation of renal lipids.

Membrane solubilization in erythrocytes as a measure of radiation exposure to fast neutrons

Membrane solubilization and osmotic fragility of rat erythrocytes irradiated in vivo with fast neutron fluences ranging from to using a source were measured instantaneously using a light scattering technique. The solubilization of erythrocyte membrane by a non-ionic detergent, octylglucoside (OG), was found to exhibit a two stage transition from vesicular form to mixed micellar form in the range of detergent concentrations 1.5-7.8 mM. The coexistence phase, vesicular/mixed micellar, was shifted towards higher detergent concentrations with increase in the neutron fluence, indicating increasing membrane resistance to the detergent and hence change in the natural membrane permeation properties. The technique shows an adequate sensitivity in detecting membrane damage in erythrocytes and has potential as a biophysical marker of radiation exposure. The osmotic fragility of irradiated erythrocytes shows a decreasing trend with increasing irradiation fluence measured directly and two weeks post-irradiation. Blood films photographed two weeks post-irradiation show developed elliptocytosis and crenated cell anaemia.

Phospholipid hydroperoxide peroxidase activities in erythrocytes.

Erythrocytes have a high oxygen content which is in repeated flux, and possess membranes rich in polyunsaturated fatty acids that are highly susceptiile to oxidative damage during their life span in ckdation [13]. In addition, haemoglobii (Hb) can react with preexisting lipid hydroperoxides to yield radical-mediated reactions that produce extensive lipid peroxidation [4]. Protection against oxidative damage of erythrocyte membranes and h o l y s i s are aEorded by vitamin E, selenium, glutathione and glutathione peroxidase (GPx) [57. However, the protection against oxidative damage to cell membrane lipids mediated by the reduction of endogenously formed lipid hydroperoxides to hydroxy derivatives through intradular GPx has been unconvincing [8], since GPX is not directly active on hydroperoxy-phospholipids which are the primary products in peroxidized biomembranes [9]. We have showed previously that hydroperoxy-phospholipids are mainly metabolised by a seleniumdependent phospholipid hydroperoxide glutathione peroxidase (PHGPX) in cultured cells [lo]. Although there are no reports about the presence of PHGPx in erythrocytes, it has been suggested that PHGPx in red blood cells may be inadequate [6]. In this paper, a PHGPx-like activity in erythrocyte membranes has been detected which was Wer investigated using immuno blotting.

Protection against oxidative damage of erythrocyte membrane by the flavonoid quercetin and its relation to iron chelating activity

Incubation of glutathione (GSH) depleted mouse erythrocytes with the oxidants phenylhydrazine, acrolein, divicine and isouramil resulted in the release of free iron and in lipid peroxidation and hemolysis. The addition of the flavonoid quercetin, which chelates iron and penetrates erythrocytes, resulted in remarkable protection against lipid peroxidation and hemolysis. The protection seems to be due to intracellular chelation of iron, since a semi-stoichiometric ratio between released iron and the amount of quercetin necessary to prevent lipid peroxidation and hemolysis was found. Incubation of GSH depleted human erythrocytes with divicine and isouramil did not induce lipid peroxidation and hemolysis in spite of a substantial release of iron. However, divicine and isouramil produced alterations of membrane proteins, such as spectrin and band 3, as well as formation of senescent cell antigen. The addition of quercetin prevented these alterations.

Cadmium-Induced Lipid Peroxidation and the Antioxidant System in Rat Erythrocytes : the Role of Antioxidants

Cadmium (Cd)-induced oxidative damage in erythrocytes causes loss of membrane function by enhancing lipid peroxidation (LPO) and altering the erythrocyte antioxidant system. Vitamin E and/or selenium (Se) was administered to rats, prior to Cd intoxication, in order to prepare the animals to withstand oxidative assault. The treatment with Cd increased LPO in erythrocytes while animals pretreated with vitamin E and/or Se prior to Cd treatment showed decreased LPO as compared with animals given Cd alone. The erythrocyte SOD and CAT activities decreased significantly with Cd treatment. The pretreatment with vitamin E and/or Se prior to Cd administration partially reversed such changes. The erythrocytes showed a marked depletion in glutathione (GSH) content with Cd treatment. The antioxidant treatments before Cd administration helped to maintain the erythrocyte GSH content. The erythrocyte glutathione reductase (GSH-R) activity increased markedly when treatments with vitamin E and Se were applied. The GSH-R activity was not observed to decrease in animals treated with antioxidant prior to Cd intoxication, which may mean that the replenishment of erythrocyte GSH content is via GSH-R. The glutathione-S-transferase (GST) activity increased significantly with Cd intoxication; however, treatment with antioxidants prior to Cd treatment decreased erythrocyte GST activity. The results show that Cd-induced LPO decreased the antioxidant capability of the erythrocytes, causing erythrocyte membrane damage.

Molecular mechanism of drug photosensitization: VIII. Effect of inorganic ions on membrane damage photosensitized by naproxen

The inhibitory effect of Cu 2+, Mn 2+, Co 2+, and I − on naproxen-induced photohemolysis was investigated. In order to better understand this effect, these ions were also tested for lipid peroxidation and protein crosslinking, which are among the main processes involved in erythrocyte membrane damage. The overall results support the hypothesis that metal cations act via a redox scavenging of the radicals which are produced on the lipid component of the membrane. This process occurs through hydrogen abstraction operated by photogenerated naproxen radicals. Moreover, copper can also act as a superoxide anion scavenger: its decay is noxious in photohemolysis, whereas it is not in lipid peroxidation. Metal cations, besides, are not able to scavenge protein crosslinking. On the other hand, iodide is able to reduce both processes because it acts as a heavy atom, favoring intersystem crossing to the unreactive triplet state of the drug, thus reducing naproxen photolysis and, as a consequence, the amount of the damaging species produced. This mechanism was supported by luminescence experiments performed in the absence and in the presence of iodide.

Reduced susceptibility to oxidative damage of erythrocyte membranes from medicated schizophrenic patients

Susceptibility to non-enzymatic peroxidation of erythrocyte membranes from medicated schizophrenic patients relative to healthy control subjects was investigated by measuring internalization into erythrocytes of ethylene glycol, cellobiotol or mannitol, with or without preincubation with cumene hydroperoxide or with chlorpromazine. The main finding was that erythrocytes from schizophrenic patients were less susceptible than those from control subjects to non-enzymatic oxidative damage from cumene hydroperoxide, as measured by internalization of cellobiotol and mannitol. At baseline before incubation, there was reduced internalization of cellobiotol and mannitol and this was reduced even further by preincubation with chlorpromazine. It is suggested that previous findings of fatty acid deficits in erythrocyte membranes from neuroleptic-treated schizophrenic patients are unlikely to have resulted from non-enzymatic oxidative damage of the membrane. Furthermore, it is suggested that depleted erythrocyte membrane essential fatty acids are more likely to be the result of the schizophrenic process rather than antipsychotic drug treatment, and that antipsychotic drugs may even offer protection against membrane lipid peroxidation.

Changes in Erythrocyte Membrane Lipids in Breast Cancer after Radiotherapy and Chemotherapy

CMF and radiation treatment alter the erythrocyte membrane architecture in breast cancer patients by inducing free radical formation in the system. The increased free radicals hydrolyze the unsaturated fatty acyl chains of membrane phospholipids and thus alter the membrane lipid profile. To assess the erythrocyte membrane damage caused by CMF chemotherapy and radiation treatment among breast cancer patients and to find a way which can protect them from the observed toxic manifestations form the basis of this study.

Plasmodium falciparum: Role of Activated Blood Monocytes in Erythrocyte Membrane Damage and Red Cell Loss during Malaria

The role of Plasmodium falciparum and blood monocytes in the erythrocyte damage and pathogenesis of anemia has been investigated using two strains of the parasite; one laboratory-established strain (FSJ-M) and one wild, fresh, clinical isolate (PfPGI). Peripheral blood monocyte-induced growth inhibition of the parasites, erythrocyte membrane lipid peroxidation as seen by the formation of lipid peroxide products, and sensitivity to peroxide hemolysis at atmospheric oxygen were evaluated. The growth inhibition of FSJ-M by activated blood monocytes was greater than that of PfPGI. The extent of lipid peroxidation and sensitivity to hemolysis increased significantly as the parasites matured. These adverse effects were more marked following exposure to activated monocytes, especially in synchronized, parasitized RBCs. In addition, uninfected erythrocytes within the PfPGI parasite culture revealed a significant increase in the lipid peroxide formation (P < 0.01) and susceptibility to lysis (P < 0.05) under similar oxidant stress induced by monocytes from normal healthy donors. Furthermore, there was a direct correlation between membrane lipid peroxidation and peroxide hemolysis, both before and after monocyte exposure, suggesting a primary role of membrane peroxidation in red cell lysis. The contribution of intraerythrocytic parasites and nonspecific activation of blood monocytes in the pathophysiology of erythrocyte damage and anemia of P. falciparum infection is discussed.

Photophysical Alterations of Erythrocyte Membrane by 4,5′,8-Trimethylpsoralen: Membrane Response Towards Psoralen Phototherapy

Despite the wide and versatile use of psoralen phototherapy in various dermal diseases including psoriasis and leukoderma, the proposed mechanism of action of psoralen is disputed. This study launches an original documentation of free-radical resulting in erythrocyte membrane damage studied by spin-labeling using the probes malnet and 5 doxyl stearate as protein and lipid domains, respectively. Both Type I and Type II photodynamics appear plausible and the therapeutic aspects of this innovation are being verified.