Red Blood Cell Glutathione Levels Research Articles

Hesperetin-7-Rutinoside Mitigates Erythrocyte Dyslipidemia and Oxidative Stress Induced by Dichlorvos

In many rural and semi-urban areas of Nigeria, dichlorvos (DDVP) is a commonly utilized pesticide. However, it provokes remarkable health hazards, including erythrocyte toxicity. Hesperetin-7-rutinoside (Hesp-7-Rut), a citrus flavanone glycoside with antioxidant, anti-dyslipidemia and acetylcholinesterase-modulating properties has been investigated for its protective effects against various xenobiotic-mediated toxicities. This inquiry probed the chemotherapeutic role of Hesp-7-Rut in mitigating erythrocyte damage in rats. Forty-two rats were randomly `apportioned into seven groups (6 rats/group): control, DDVP alone (8 mg.kg⁻¹day⁻¹), DDVP + Hesp-7-Rut (50 mg.kg⁻¹day⁻¹), DDVP + Hesp-7-Rut (100 mg.kg⁻¹day⁻¹), DDVP + atropine (0.2 mg.kg⁻¹day⁻¹), Hesp-7-Rut only (50 mg.kg⁻¹day⁻¹), and Hesp-7-Rut only (100 mg.kg⁻¹day⁻¹). Rats were orally administered DDVP for 7 days followed by Hesp-7-Rut or atropine for another 14 days. Blood samples were collected to assess for biochemical assays. Hesp-7-Rut significantly (p<0.05) rescinded DDVP-prompted increases in erythrocyte nitric oxide, malondialdehyde, cholesterol, phospholipids, and cholesterol: phospholipids ratio. Additionally, Hesp-7-Rut reversed DDVP-elicited decreases in red blood cell glutathione levels, activities of GST, SOD, catalase, glutathione peroxidase, and acetylcholinesterase. Overall, Hesp-7-Rut efficiently counteracts DDVP-elicited erythrocyte dysfunction by mitigating oxidative stress, dyslipidemia, and acetylcholinesterase inhibition. These findings highlight the potential of Hesp-7-Rut as a promising therapeutic agent for mitigating the harmful effects of DDVP exposure.

Total body irradiation causes a chronic decrease in antioxidant levels

Ionizing radiation exposure may not only cause acute radiation syndrome, but also an increased risk of late effects. It has been hypothesized that induction of chronic oxidative stress mediates the late effects of ionizing radiation. However, only a few reports have analyzed changes in long-term antioxidant capacity after irradiation in vivo. Our previous study demonstrated changes in whole-blood antioxidant capacity and red blood cell (RBC) glutathione levels within 50 days after total body irradiation (TBI). In this study, seven-week-old, male, C57BL/6J mice exposed to total body irradiation by X-ray and changes in whole-blood antioxidant capacity and RBC glutathione levels at ≥ 100 days after TBI were investigated. Whole-blood antioxidant capacity was chronically decreased in the 5-Gy group. The RBC reduced glutathione (GSH) level and the GSH/oxidative glutathione (GSSG) ratio were chronically decreased after ≥ 1 Gy of TBI. Interestingly, the complete blood counts (CBC) changed less with 1-Gy exposure, suggesting that GSH and the GSH/GSSG ratio were more sensitive radiation exposure markers than whole-blood antioxidant capacity and CBC counts. It has been reported that GSH depletion is one of the triggers leading to cataracts, hypertension, and atherosclerosis, and these diseases are also known as radiation-induced late effects. The present findings further suggest that chronic antioxidant reduction may contribute to the pathogenesis of late radiation effects.

Effect of Sintering Temperature of Bioactive Glass Nanoceramics on the Hemolytic Activity and Oxidative Stress Biomarkers in Erythrocytes.

The nature of the surface is critical in determining the biological activity of silica powders. A novel correlation between toxicity and surface properties of bioactive glass ceramics (BGCs) synthesized via the sol-gel method was attempted in this study. The behavior of BGCs after their attachment to the surface of red blood cells (RBCs) was evaluated and their toxic effects were determined based on hemolysis, membrane injury via anti-phosphotyrosine immunoblot of Band 3, lipid peroxidation, potential to generate reactive oxygen species, and antioxidant enzyme production. In particular, three BGCs were synthesized and treated at three sintering temperatures (T1 = 835°C, T2 = 1000°C and T3 = 1100°C) to investigate possible relation between surface charge or structure and hemolytic potential. Their toxicity based on hemolysis was dose dependent, while BGC-T2 had the best hemocompatibility in compare with the other BGCs.No BGCs in dosages lower than 0.125mg/mL could damage erythrocytes. On the other hand, all BGCs promoted the production of reactive oxygen species in certain concentrations, with the BGC-T2 producing the lowest ROS and increasing the glutathione levels in RBCs protecting their damage. The results suggest that various factors such as size, a probable different proportion of surface silanols, a balanced mechanism between calcium and magnesium cellular uptake or the different crystalline nature may have contributed to this finding; however, future research is needed to clarify the underlying mechanisms.

Pilot clinical study of ascorbic acid treatment in cardiac catheterization.

Clinical radiodiagnosis and radiotherapy sometimes induce tissue damage and/or increase the risk of cancer in patients. However, in radiodiagnosis, a reduction in the exposure dose causes a blockier image that is not acceptable for diagnosis. Approximately 70% of DNA damage is induced via reactive oxygen species and/or radicals created during X-ray irradiation. Therefore, treatment with anti-oxidants and/or radical scavengers is considered to be effective in achieving a good balance between image quality and damage. However, few studies have examined the effect of using radical scavengers to reduce radiation damage in the clinical setting. In this study, we administrated 20 mg/kg ascorbic acid (AA) to patients before cardiac catheterization (CC) for diagnostic purposes. We analyzed changes in the number of phosphorylated H2AX (γH2AX) foci (a marker of DNA double-strand breaks) in lymphocytes, red blood cell glutathione levels, blood cell counts, and biochemical parameters. Unfortunately, we did not find satisfactory evidence to show that AA treatment reduces γH2AX foci formation immediately after CC. AA treatment did, however, cause a higher reduced/oxidized glutathione ratio than in the control arm immediately after CC. This is a preliminary study, but this result suggests that reducing radiation damage in clinical practice can be achieved using a biological approach.

The Severity of Autism Is Associated with Toxic Metal Body Burden and Red Blood Cell Glutathione Levels

This study investigated the relationship of children's autism symptoms with their toxic metal body burden and red blood cell (RBC) glutathione levels. In children ages 3–8 years, the severity of autism was assessed using four tools: ADOS, PDD-BI, ATEC, and SAS. Toxic metal body burden was assessed by measuring urinary excretion of toxic metals, both before and after oral dimercaptosuccinic acid (DMSA). Multiple positive correlations were found between the severity of autism and the urinary excretion of toxic metals. Variations in the severity of autism measurements could be explained, in part, by regression analyses of urinary excretion of toxic metals before and after DMSA and the level of RBC glutathione (adjusted R 2 of 0.22–0.45, P < .005 in all cases). This study demonstrates a significant positive association between the severity of autism and the relative body burden of toxic metals.

Effects of N-Acetyl-Cysteine on Endothelial Function and Inflammation in Patients with Type 2 Diabetes Mellitus

Endothelial dysfunction has been associated with premature vascular disease. There is increasing data that N-acetyl-cysteine (NAC) may prevent or improve endothelial dysfunction. The aim of this study was to assess the effects of NAC on endothelial function in patients with type 2 diabetes mellitus, a population at high risk for endothelial dysfunction. Twenty-four patients with diabetes mellitus were assigned randomly to initial therapy with either 900 mg NAC or placebo twice daily in a double-blind, cross-over study design. Flowmediated vasodilation (FMD) of the brachial artery was assessed at baseline, after four weeks of therapy, after a four-week wash-out period, and after another four weeks on the opposite treatment. Plasma and red blood cell glutathione levels and high-sensitivity C-reactive protein (CRP) were measured at all four visits. At baseline, FMD was moderately impaired (3.7±2.9%). There was no significant change in FMD after four weeks of NAC therapy as compared to placebo (0.1±3.6% vs. 1.2±4.2%). Similarly, there was no significant change in glutathione levels. However, median CRP decreased from 2.35 to 2.14 mg/L during NAC therapy (p=0.04), while it increased from 2.24 to 2.65 mg/L with placebo. No side effects were noted during the treatment period. In this double-blind, randomized cross-over study, four weeks of oral NAC therapy failed to improve endothelial dysfunction in patients with diabetes mellitus. However, NAC therapy decreased CRP levels, suggesting that this compound may have some efficacy in reducing systemic inflammation.

Inhibition of neutral endopeptidase potentiates neutrophil activation during Mg-deficiency in the rat.

Neutral endopeptidase (NEP), which degrades substance P (SP), may regulate neutrophil activation during Mg-deficiency (MgD). Male Sprague-Dawley rats (180g) were fed MgD (approximately 50 mg Mg/kg) or Mg-sufficient (MgS, 608 mg Mg/kg) diets for 7 days +/- NEP inhibitor phosphoramidon (PR, 5 mg/kg/day, s.c.). MgD alone induced a 9-fold (vs. MgS, p <0.01) elevation in plasma SP; MgD+PR enhanced it further to 18-fold (p <0.001). Neutrophils from MgD+PR rats displayed a 3.9-fold higher (p <0.01) basal .O(2-) generation, but those from MgD or PR alone were not activated. Plasma PGE2-metabolite levels rose 2.67- (p <0.01) and 1.56- (p <0.05) fold, respectively, in MgD+PR and MgD groups; the corresponding red blood cell glutathione levels were decreased 21% (p <0.025) and 7% (NS). MgD+PR significantly reduced neutrophil NEP activity by 48% (p <0.02); PR or MgD alone only reduced this activity 26% and 15%, respectively. We conclude that NEP inhibition potentiates SP-mediated neutrophil .O(2-) production and may promote other inflammatory activities during MgD.

In vivo and in vitro investigations of the effects of the antimalarial drug dihydroartemisinin (DHA) on rat embryos

Artemisinin derivatives are clinically effective and safe antimalarials, but are not recommended during the first trimester of pregnancy because of the resorptions and abnormalities seen in animal reproduction studies. Understanding how, when and what toxicity occurs is crucial to any assessment of clinical relevance. Previously, DHA has been shown in the rat whole embryo culture (WEC) to primarily affect primitive red blood cells (RBCs) causing subsequent tissue damage and dysmorphogenesis. To verify the primary target of DHA in vivo and to detect consequences induced by early damage on embryo development, pregnant female rats were orally treated on gestation days (GD) 9.5 and 10.5 with 7.5 or 15 mg/kg/day DHA and caesarean sectioned on GD11.5, 12.5, 13.5, 15 and 20. A parallel in vitro WEC study evaluated the role of oxidative damage and examined blood islands and primitive RBCs. In accordance with the WEC results, primitive RBCs from yolk sac hematopoiesis were the target of DHA in vivo. The resulting anemia led to cell damage, which depending on its degree, was either diffuse or focal. Embryonic response to acute anemia varied from complete recovery to malformation and death, depending on the extent of cell death. Malformations occurred only in litters with embryonic deaths. DHA induced low glutathione levels in RBCs, indicating that oxidative stress may be involved in artemisinin toxicity; effects were extremely rapid, with altered RBCs seen as early as GD10. In establishing the relevance of these findings to humans, one should consider differences in the development of rodents and humans. While yolk sac hematopoiesis occurs similarly in the two species, early placentation and extent of exposure differ. In particular, early hematopoiesis takes only 7 days in rats (during which RBCs expand in a clonal fashion) compared with 6 weeks in humans; thus the susceptible period in relation to the duration of exposure to an artemisinin-based treatment may be substantially different.

The effects of hormone replacement therapy combined with vitamins C and E on antioxidants levels and lipid profiles in postmenopausal women with Type 2 diabetes*1

Background: Recent studies have demonstrated that oxidative modification of low-density lipoprotein (LDL) involving the formation of lipid peroxides (MDA), exerts several biological effects that may contribute to the onset and progression of cardiovascular diseases in postmenopausal women with Type 2 diabetes (DPMW). Therefore, the aim of our study was to evaluate the effect of hormone replacement therapy (HRT), vitamin C and E (VCE) treatments on lipid profiles, glucose and MDA levels as well as antioxidant vitamins and enzymes in plasma and red blood cells (RBC) in diabetic or non-diabetic postmenopausal women (PMW). Methods: Oral HRT and VCE supplementation for 6 weeks were compared with HRT treatment in 40 non-diabetic PMW and 40 DPMW. Results: In the 40 postmenopausal women (PMW) and 20 postmenopausal women with DPMW who received oral HRT and 20 DPMW who received HRT plus VCE, there was a significant fall in MDA, total cholesterol, LDL-cholesterol and triglyceride values. Glycated haemoglobin (HbA1c) in the DPMW was significantly improved with oral HRT and VCE although no significant change in white blood cell counts, vitamin A and HDL values occurred. Additionally, a fall in plasma glucose, HbA1c and platelet values also occurred in the PMW and DPMW groups by oral HRT and VCE treatments. There was a significant increase in plasma vitamin E and beta-carotene concentrations, catalase, glutathione peroxidase and reduced glutathione levels in RBC and plasma in DPMW by treatments with HRT and/or VCE. Conclusions: Daily VCE and HRT administrations both in PMW and DPMW seem to produce significant improvement in antioxidants concentrations, and the metabolic control of lipids and glucose. The HRT and VCE supplementations may strengthen the antioxidant defense system due to reducing blood glucose and lipid metabolites, and they may play a role in preventing cardiovascular diseases in postmenopausal women with Type 2 diabetes.

The effects of hormone replacement therapy combined with vitamins C and E on antioxidants levels and lipid profiles in postmenopausal women with Type 2 diabetes

Background: Recent studies have demonstrated that oxidative modification of low-density lipoprotein (LDL) involving the formation of lipid peroxides (MDA), exerts several biological effects that may contribute to the onset and progression of cardiovascular diseases in postmenopausal women with Type 2 diabetes (DPMW). Therefore, the aim of our study was to evaluate the effect of hormone replacement therapy (HRT), vitamin C and E (VCE) treatments on lipid profiles, glucose and MDA levels as well as antioxidant vitamins and enzymes in plasma and red blood cells (RBC) in diabetic or non-diabetic postmenopausal women (PMW). Methods: Oral HRT and VCE supplementation for 6 weeks were compared with HRT treatment in 40 non-diabetic PMW and 40 DPMW. Results: In the 40 postmenopausal women (PMW) and 20 postmenopausal women with DPMW who received oral HRT and 20 DPMW who received HRT plus VCE, there was a significant fall in MDA, total cholesterol, LDL-cholesterol and triglyceride values. Glycated haemoglobin (HbA1c) in the DPMW was significantly improved with oral HRT and VCE although no significant change in white blood cell counts, vitamin A and HDL values occurred. Additionally, a fall in plasma glucose, HbA1c and platelet values also occurred in the PMW and DPMW groups by oral HRT and VCE treatments. There was a significant increase in plasma vitamin E and beta-carotene concentrations, catalase, glutathione peroxidase and reduced glutathione levels in RBC and plasma in DPMW by treatments with HRT and/or VCE. Conclusions: Daily VCE and HRT administrations both in PMW and DPMW seem to produce significant improvement in antioxidants concentrations, and the metabolic control of lipids and glucose. The HRT and VCE supplementations may strengthen the antioxidant defense system due to reducing blood glucose and lipid metabolites, and they may play a role in preventing cardiovascular diseases in postmenopausal women with Type 2 diabetes.

Effects of vitamin E supplementation on blood antioxidants levels in patients with Behçet’s disease

Behçet’s disease (BD) is known for many years, yet its etiology remains unknown. In BD, the increased production of reactive oxygen species from activated neutrophils may reduce concentrations of antioxidant vitamins and enzymes in plasma and red blood cells (RBC). Vitamin E is an important fat soluble antioxidant and its role on antioxidant parameters of BD is unclear. The study was undertaken to evaluate the role of vitamin E on lipid peroxidation (MDA) levels, antioxidant vitamin and enzyme concentrations in plasma and RBC in patients with BD. There were three groups i.e., control, patient and treatment groups with twenty-five subjects in each. Nonsmoking patients with BD, patient group, was compared with an equal number of healthy control subjects (control group). Blood samples were taken from both control and patient groups and then oral vitamin E was daily supplemented to the patients with BD for six weeks (treated group). At the end of six weeks, blood was taken from the treated group once more. RBC and plasma MDA levels, serum neopterin, complement system (C 3 and C 4), ASO, CRP, rheumatoid factor, plasma lipoprotein (a), total cholesterol concentrations and erythrocyte sedimentation rate were higher in the patient group than in the control group, but they were lower in the treatment group than in the patient group. While vitamins A, E and β-carotene concentrations in plasma, catalase, glutathione peroxidase activities and reduced glutathione levels in RBC and plasma were lower in patient group than in the controls, they were found to be higher in the treatment group than in the patient group. These results provide some evidence for a potential role of increased lipid peroxidation and decreased enzymatic and non enzymatic antioxidants in BD by its inflammatory character and vitamin E which may strengthen the antioxidant defense system, and may contribute to the treatment of BD.

The Molecular Basis of a Case of γ-Glutamylcysteine Synthetase Deficiency

Gamma-glutamylcysteine synthetase catalyzes the first step in glutathione synthesis. The enzyme consists of 2 subunits, heavy and light, with the heavy subunit serving as the catalytic subunit. A patient with hemolytic anemia and low red blood cell glutathione levels was found to have a deficiency of gamma-glutamylcysteine synthetase activity. Examination of cDNA from the patient and her mother showed that she was homozygous and that her mother was heterozygous for a A-->T transversion at nt1109 producing a deduced amino acid change of His370Leu. The partial genomic structure of the catalytic subunit of gamma-glutamylcysteine synthetase (GLCLC) was determined, providing some intron/exon boundaries to make it possible to sequence an affected part of the coding region from genomic DNA. The 1109A-->T mutation was not present in the DNA of 38 normal subjects. In the course of these studies we found a diallelic polymorphism in nt +206 of an intron and another polymorphism that consisted of a duplication of a CAGC at cDNA nt1972-1975 in the 3' untranslated region. The 2 polymorphisms were found to be only in partial linkage disequilibrium.

Effect of parathyroid hormone on antioxidant enzyme activity and lipid peroxidation of erythrocytes in five-sixths nephrectomized rats.

The purpose of the study was to investigate the effects of parathyroid hormone (PTH) infusion on antioxidant enzyme activity and lipid peroxidation of erythrocytes in five-sixths nephrectomized (Nx) rats. Five-sixths Nx rats had a higher osmotic fragility in red blood cells (RBC). Thyroparathyroidectomy (TPTX) effectively decreased the abnormality of osmotic fragility in RBC in Nx rats. PTH infusion in Nx-TPTX rats markedly increased the osmotic fragility in RBC. Total glutathione was measured by using the enzyme-recycling method. We found elevated glutathione levels in RBC of five-sixths Nx rats, but this elevation could be inhibited by TPTX and recovered by PTH infusion in Nx-TPTX rats. Five-sixths Nx rats had a lower glutathione peroxidase activity in RBC, but TPTX or PTH infusion was not found to alter the decrease of the glutathione peroxidase activity in RBC of five-sixths Nx rats. These rats had a higher activity in RBC superoxide dismutase as compared with sham-operated controls (p < 0.05), but the higher activity in RBC superoxide dismutase in Nx rats had been inhibited by TPTX. PTH infusion recovered the higher activity in RBC superoxide dismutase in five-sixths Nx-TPTX rats. Nx rats were not found to alter the activity of catalase in RBC. Neither could TPTX or PTH infusion in Nx rats influence the activity of catalase in RBC. A high lipid peroxidation in RBC was found in five-sixths Nx rats, namely, increased formation of malondialdehyde (MDA) in RBC had been induced to produce lipid peroxidation by H2O2, but neither TPTX nor PTH infusion could inhibit or enhance the increase of lipid peroxidation in RBC of Nx rats. These results indicate that PTH infusion did not increase the susceptibility to lipid peroxidation in RBC of five-sixths Nx rats. Thus, the increased osmotic fragility in RBC induced by PTH infusion may not result from the reduction in the RBC defense mechanism against free radical toxicity.

Loss of red blood cell glutathione during Mg deficiency: prevention by vitamin E, D-propranolol, and chloroquine

Mg deficiency results in loss of red blood cell glutathione and was thought to be due to decreased Mg-dependent synthesis. The effects of vitamin E, D-propranolol, and chloroquine on red blood cell glutathione levels in Mg-deficient rats were examined. Feeding the rats a Mg-deficient diet for 3 wk resulted in an approximately 80% decrease in serum Mg and a 55% loss of red blood cell glutathione; concomitantly, plasma thiobarbituric acid reactive (TBAR) materials rose 240%. All three drug treatments had no effect on the plasma Mg levels but significantly inhibited the rise in TBAR content and attenuated (60-80% effective) the loss of glutathione. Red blood cell ghost membranes from the Mg-deficient rats also exhibited 2.3-fold higher TBAR content, which was attenuated by vitamin E treatment. With isolated red blood cells from Mg-sufficient rats, loss of glutathione could be induced by a chemical oxyradical system. Direct protective effects were afforded by alpha-tocopherol and D-propranolol but not by chloroquine. The data suggest that 1) the loss of glutathione during Mg deficiency was due to increased oxidative degradation, 2) both vitamin E and D-propranolol protected by a membrane antiperoxidative action, and 3) chloroquine probably protected by diminishing prooxidant activity secondary to its inhibition of cytokine induction during Mg deficiency.

Glutathione in the red blood cells of embryonic mice

1. 1. A micro-method for the determination of red blood cell glutathione levels has been developed. 2. 2. It has been shown that, in order to obtain true values of glutathione concentrations within red blood cells, it is necessary to correct for loss of labelled glutathione and also to take account of the time taken to complete the analytical procedure. 3. 3. Glutathione is the major small molecular weight thiol present in embryonic red blood cells. 4. 4. The glutathione to haemoglobin ratio is maintained at 0.6 from 13 days gestation to adulthood in the mouse. 5. 5. Decay of glutathione in both adult and embryonic red blood cells can be avoided by incubation of the red blood cells in glucose containing buffers.