Assessment of oxidative stress in patients with sickle cell disease: The glutathione system and the oxidant–antioxidant status

Gizi, Anna; Papassotiriou, Ioannis; Apostolakou, Filia; Lazaropoulou, Christina; Papastamataki, Maria; Kanavaki, Ino; Kalotychou, Vassiliki; Goussetis, Evgenios; Kattamis, Antonios; Rombos, Ioannis; Kanavakis, Emmanuel

doi:10.1016/j.bcmd.2011.01.002

Cited by 97 publications

(99 citation statements)

References 31 publications

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“…At baseline, total GSH content (GSH ϩ GSSG) was lower in SSRBCs than in normal RBCs, as described previously by others 21 (supplemental Figure 1C-D, available on the Blood Web site; see the Supplemental Materials link at the top of the online article). Further, diminished GSH-based reducing power in SSRBCs was evidenced by a weaker (ie, less negative) reduction potential ( Figure 1C).…”

Section: Reducing Equivalent Recycling Capacitysupporting

confidence: 82%

“…Blood was drawn from healthy volunteers or from patients homozygous for HbS; 40 children with SCA were recruited during routine clinic visits. The mean values at sampling were: age: 12.4 years (range, [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22]; hematocrit: 23.3% (range, 17.2%-34.5%); and reticulocytes: 14.1% (range, 2.0-26.9). None of the patients had active painful episodes, respiratory pathology, nor had been transfused within 3 months.…”

Section: Blood Sampling Purification Of Rbc Membranes and Hemoglobinmentioning

confidence: 99%

“…6,7,[18][19][20] Consequent loss of O 2 -dependent EMP control might then reduce HMP substrate availability, limiting NADPH and GSH recycling capacity and thereby creating vulnerability to oxidative stress, an important and highly variable manifestation of the SCA phenotype. 2,21 Of the EMP enzymes under cdB3 control, we focused on GAPDH, which is notable for its location at the juncture of the HMP/EMP pathways. To address our hypothesis, we used 1 Hnuclear magnetic resonance ( 1 H-NMR) spectroscopy to precisely quantitate proportional HMP flux in intact RBCs and developed a novel, inverted membrane-on-bead model to study reciprocal binding between HbS and GAPDH on cdB3 under physiologic conditions.…”

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confidence: 99%

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Sickle hemoglobin disturbs normal coupling among erythrocyte O2 content, glycolysis, and antioxidant capacity

Rogers¹,

Ross²,

Davignon

et al. 2013

Blood

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Key Points• Hb-conformation-dependent interaction with band 3 protein regulates glycolysis in RBCs.• In hypoxia, HbS disrupts this system, disabling RBC antioxidant defense. Energy metabolism in RBCs is characterized by O 2 -responsive variations in flux through the Embden Meyerhof pathway (EMP) or the hexose monophosphate pathway (HMP). Therefore, the generation of ATP, NADH, and 2,3-DPG (EMP) or NADPH (HMP) shift with RBC O 2 content because of competition between deoxyhemoglobin and key EMP enzymes for binding to the cytoplasmic domain of the Band 3 membrane protein (cdB3). Enzyme inactivation by cdB3 sequestration in oxygenated RBCs favors HMP flux and NADPH generation (maximizing glutathione-based antioxidant systems). We tested the hypothesis that sickle hemoglobin disrupts cdB3-based regulatory protein complex assembly, creating vulnerability to oxidative stress. In RBCs from patients with sickle cell anemia, we demonstrate in the present study constrained HMP flux, NADPH, and glutathione recycling and reduced resilience to oxidative stress manifested by membrane protein oxidation and membrane fragility. Using a novel, inverted membrane-on-bead model, we illustrate abnormal (O 2 -dependent) association of sickle hemoglobin to RBC membrane that interferes with sequestration/inactivation of the EMP enzyme GAPDH. This finding was confirmed by immunofluorescent imaging during RBC O IntroductionSickle cell anemia (SCA) arises from a single amino acid substitution (Glu6Val) in the ␤-globin chain. Although the change to hemoglobin (Hb) is simple and uniform, SCA is characterized by broad differences in clinical manifestation. Phenotype variation in SCA is thought to arise from both environmental and genetic factors (eg, ␤-gene cluster haplotype, degree of HbF expression, or effects of other epistatic genes). The environmental factor that most clearly influences SCA phenotype is hypoxia, which drives sickle Hb (HbS) polymerization and the resulting well-characterized alterations in RBC physiology and the microcirculation. However, the influence of hypoxia on the SCA phenotype appears to be insufficiently explained by HbS polymerization alone. 1 Moreover, we lack a clear mechanistic understanding of the significant oxidative stress complicating SCA, a key feature of phenotype variation, both at rest and in association with hypoxia. 2 Nonpolymerized, solution-phase HbS may promote oxidative stress, even in RBCs under normal physiologic O 2 gradients. 3 Specifically, the low redox potential for heme in HbS 4 and avid binding affinity of HbS for the cytoplasmic regulatory domain of the Band 3 membrane protein (cdB3) 5,6 strongly affect RBC energetics and antioxidant systems [7][8][9] and, notably, do so as a function of RBC O 2 content. Therefore, both the genesis and the disposal of reactive oxygen species are abnormal in SCA, creating a baseline state of oxidative stress, which worsens in hypoxia.In particular, consideration of metabolic control in RBCs suggests O 2 -dependent HbS-cdB3 interaction as a relatively ...

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Section: Reducing Equivalent Recycling Capacitysupporting

confidence: 82%

Section: Blood Sampling Purification Of Rbc Membranes and Hemoglobinmentioning

confidence: 99%

mentioning

confidence: 99%

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Sickle hemoglobin disturbs normal coupling among erythrocyte O2 content, glycolysis, and antioxidant capacity

Rogers¹,

Ross²,

Davignon

et al. 2013

Blood

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“…Moreover, NADPH oxidase activity may deplete the cellular pool of NADPH, thus impairing the ability of the RBC to maintain its antioxidant defenses. 2,13 In addition to inducing endogenous damage, erythrocyte-derived ROS, which can exit the cell via the membrane anion channel, 25 may also contribute to systemic oxidative stress by modifying the activity of plasma proteins, WBC, platelets, and endothelial cells. 1 The potential utility of antioxidant therapy in SCD is intriguing but with limited evidence to date that it can ameliorate the acute or chronic pathophysiology of the disease.…”

Section: Nadph Oxidase and Ros In Sickle Erythrocytes 2105mentioning

confidence: 99%

“…[3][4][5] RBC and other cell types show evidence of lipid peroxidation and oxidative damage to structural proteins. [6][7][8] Additionally, plasma from SCD patients has elevated levels of advanced glycation end products 9,10 and products of lipid peroxidation (F-2 isoprostanes, malonaldehyde, and 4-hydroxynonenal), [11][12][13] all of which are markers of oxidative stress. There are several postulated mechanisms for the increased oxidative stress in patients with SCD.…”

Section: Introductionmentioning

confidence: 99%

Erythrocyte NADPH oxidase activity modulated by Rac GTPases, PKC, and plasma cytokines contributes to oxidative stress in sickle cell disease

George

Pushkaran

Konstantinidis

et al. 2013

Blood

172

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• Sickle RBC ROS production is mediated in part by NADPH oxidase activity.• Sickle RBC ROS production can be induced by plasma signaling molecules.Chronic inflammation has emerged as an important pathogenic mechanism in sickle cell disease (SCD). One component of this inflammatory response is oxidant stress mediated by reactive oxygen species (ROS) generated by leukocytes, endothelial cells, plasma enzymes, and sickle red blood cells (RBC). Sickle RBC ROS generation has been attributed to sickle hemoglobin auto-oxidation and Fenton chemistry reactions catalyzed by denatured heme moieties bound to the RBC membrane. In this study, we demonstrate that a significant part of ROS production in sickle cells is mediated enzymatically by NADPH oxidase, which is regulated by protein kinase C, Rac GTPase, and intracellular Ca 21 signaling within the sickle RBC. Moreover, plasma from patients with SCD and isolated cytokines, such as transforming growth factor b1 and endothelin-1, enhance RBC NADPH oxidase activity and increase ROS generation. ROS-mediated damage to RBC membrane components is known to contribute to erythrocyte rigidity and fragility in SCD. Erythrocyte ROS generation, hemolysis, vaso-occlusion, and the inflammatory response to tissue damage may therefore act in a positive-feedback loop to drive the pathophysiology of sickle cell disease. These findings suggest a novel pathogenic mechanism in SCD and may offer new therapeutic targets to counteract inflammation and RBC rigidity and fragility in SCD. (Blood. 2013;121(11):2099-2107 IntroductionVaso-occlusion and hemolysis from the rigid and concurrently fragile red blood cells (RBC) in patients with sickle cell disease (SCD) cause a variety of acute and chronic manifestations ranging from frequent and severe painful crises to stroke and chronic organ failure. Chronic inflammation has emerged as an important pathogenic mechanism in SCD, and oxidative stress is increasingly recognized as a component of this chronic inflammatory state, inducing damage to a variety of subcellular and tissue structures. 1,2Patients with SCD have decreased plasma levels of glutathione, vitamin C, and vitamin E, presumably due to consumption by increased oxidant production. [3][4][5] RBC and other cell types show evidence of lipid peroxidation and oxidative damage to structural proteins.6-8 Additionally, plasma from SCD patients has elevated levels of advanced glycation end products 9,10 and products of lipid peroxidation (F-2 isoprostanes, malonaldehyde, and 4-hydroxynonenal), [11][12][13] all of which are markers of oxidative stress. There are several postulated mechanisms for the increased oxidative stress in patients with SCD. Sickle (SS) RBC reactive oxygen species (ROS) generation has been attributed to sickle hemoglobin auto-oxidation and iron-mediated Fenton chemistry reactions catalyzed by denatured heme moieties bound to the RBC membrane.14 Plasma hemoglobin and free heme resulting from chronic hemolysis generate superoxide radicals via the same nonenzymatic mechanisms. 15 In...

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Antioxidant supplementation for sickle cell disease

Bolarinwa

Oduwole

Okebe

et al. 2020

Cochrane Database of Systematic Reviews

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Assessment of oxidative stress in patients with sickle cell disease: The glutathione system and the oxidant–antioxidant status

Cited by 97 publications

References 31 publications

Sickle hemoglobin disturbs normal coupling among erythrocyte O2 content, glycolysis, and antioxidant capacity

Sickle hemoglobin disturbs normal coupling among erythrocyte O2 content, glycolysis, and antioxidant capacity

Erythrocyte NADPH oxidase activity modulated by Rac GTPases, PKC, and plasma cytokines contributes to oxidative stress in sickle cell disease

Antioxidant supplementation for sickle cell disease

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