Printer Friendly

Oxidants and antioxidants of erythrocytes.

INTRODUCTION

There are three phases in the development of cells in the red-blood-cell/erythrocyte system: production and maturation of erythroblasts and reticulocytes in the bone marrow transport of mature erythrocytes in the circulating blood, and aging as well as elimination of erythrocytes from the blood by mononuclear phagocytes of the spleen. Erythroblasts posses nuclei, mitochondria and ribosomes and they are the place of hemoglobin biosynthesis.

Reticulocytes, on the other hand, do not poss nuclei, but they contain fragments of mitochondria and ribosomes, whose quantity decreases up to disappearance together with maturation [1]. They synthesize only small amounts of hemoglobin. Reticulocytes comprise only 0.030.15% of mature erythrocytes in the circulating blood.

Erythrocytes are dominant among blood cells, and their ratio to blood platelets and leukocytes is 700:40:1. There are 4-5 million of erythrocytes in 1 [mm.sup.3]. Erythrocytes total count in the circulating blood equals approximately 25 billions. Due to the structure, chemical composition and metabolism, erythrocytes are adapted to oxygen and carbon dioxide transport. Energy, stored as ATP, is obtained by anaerobic glycolysis (90%) and pentose cycle (10%). Glycolysis provides reduced nicotinamide adenine dinucleotide (NADH) and pentose cycle--reduced nicotinamide adenine dinucleotide phosphate (NADPH), essential for, among others, reduction of methemoglobin and oxidized glutathione.

Hemoglobin and heme iron

The mass of one erythrocyte is 82.8pg. Hemoglobin is a dominant chemical component of erythrocytes and comprises 33% of total mass, 94% of dry mass, and 97% of protein mass of the erythrocyte (Tab. 1).

One erythrocyte contains 32 x [10.sup.-12]g and all erythrocytes of the human body contain approximately 800g of hemoglobin. Hemoglobin appears in two conformational forms: relaxed (R) of high oxygen affinity and tense (T) of low oxygen affinity [3].

Only 12.5[cm.sup.3] of oxygen is dissolved in 5 L of blood in temperature of 37[degrees]C. Hemoglobin in this volume of blood binds as much as 1000[cm.sup.3] of oxygen, i.e. 80 times more than is physically dissolved. Thus, 1g of hemoglobin binds 1.34[cm.sup.3] and 1g of heme iron--binds 400[cm.sup.3] of oxygen. In arterial blood there is 95-98% of oxygenated hemoglobin and in venous blood 67-75% of oxygenated hemoglobin. The relaxing organism demand for oxygen at rest is 250[cm.sup.3]/min. During physical exercise, it is up to 10 fold higher, i.e. it increases to 2500[cm.sup.3]/min.

Deoxygenated/oxygenated hemoglobin, under the influence of specific chemical agents, is changed into derivatives: methemoglobin, S-nitrosohemoglobin, carbonous oxide hemoglobin, sulphhemoglobin, cyanomethemoglobin [3]. These derivatives are incapable of carrying oxygen. Some of them, specifically methemoglobin and S-nitrosohemoglobin, play a significant role in oxidative/antioxidative processes.

Hemoglobin in erythrocytes contains approximately 2.6g of iron, which comprises 60% of systemic organism iron. The organism requires 20-36mg of iron for hemoglobin synthesis per 24 hours. It comes mainly from used up erythrocytes and tissue reserves and only to a small extent from iron consumed with food. Thus, iron balance seems to be a closed system to a large extent.

The content of hemoglobin and its derivatives, heme iron and cation iron [Fe.sup.2+]/[Fe.sup.3+], with the domination of trivalent iron, is increased locally, e.g. in inflammatory focuses, in blood extravasated to tissues and body cavities, at the site of hemoglobin infiltration to the walls of arterial and venous vessels and arterial and venous thrombi [4, 40, 43, 65].

Reactive oxygen species

Transport of significant amounts of oxygen by erythrocytes favors production of reactive oxygen species. They occur in forms of superoxide anion ([O.sub.2.sup.-]), chargeless radicals, such as hydroxyl radical (OH) and reactive oxygen species such as hydrogen peroxide ([H.sub.2][O.sub.2]) and singlet oxygen ([sup.1][O.sub.2]). The oxidation reaction of [Fe.sup.2+] of hemoglobin to [Fe.sup.3+] of methemoglobin is the main source of superoxide anion in erythrocytes. One electron is transferred from ferrous cation of an oxyhemoglobin molecule to molecular oxygen (Fig.1).

Bivalent iron is oxidized to trivalent iron and an unstable indirect compound is produced--superoxide anion of methemoglobin. It undergoes dissociation to produce methemoglobin and superoxide anion:

Hb-[Fe.sup.2+] - [O.sub.2] [??] MetHb-[Fe.sup.3+] - [O.sub.2] [??] MetHb-[Fe.sup.3+] + [O.sub.2.sup.*-]

Approximately, 1.2% of hemoglobin undergoes this change on physiological conditions. In twenty four hours, about 10 x [10.sup.6] molecules of superoxide anion are produced in one erythrocyte and 250 x [10.sup.15] in all erythrocytes of the human body [5]. Production of superoxide anion is increased in congenital methemoglobinemia and methemoglobin reductase deficiency as well as acquired methemoglobinemia, e.g. after sulfonamides and antimalarial drugs and in nitrite compounds and aniline intoxication, after hemodialysis, ultrasound effect, thermal shock, and physical exercises. The rate of forming superoxide anion increases together with the increase in acidity of the environment and in the presence of anions in the sequence of: [Cl.sup.-] < [F.sup.-] < OC[N.sup.-] < SCN < [N.sub.3.sup.-] < C[N.sup.-] [6].

Not only, the superoxide anion reduces ferric cations to ferrous cations [7]:

[Fe.sup.3+] + [O.sup.2*-] [right arrow] [Fe.sup.2+] + [O.sub.2]s

but also ascorbate, NADH, NADPH, glutathione, cysteine, and protein-thiolic groups.

Hydroxide radical is formed in the Fenton reaction, in which hydrogen peroxide is reduced under the influence of ferrous cation [8]

[MATHEMATICAL EXPRESSION NOT REPRODUCIBLE IN ASCII]

The formation of hydroxyl radical in the Fenton reaction is locally specific [9]. The heme ring containing cations [Fe.sup.2+] is situated at the bottom of the hydrophobic split of alpha- and beta-globins and hydrogen peroxide, a compound well-transferable in the environment, gets there, unlike antioxidants that are no capable of getting to the alpha- and beta-globin chain. Then, a high concentration of OH and destructive action to amino-acid residues at the site of appearance take place.

Hydroxyl radical is also formed in the Haber-Weiss reaction with the participation of hydrogen peroxide, superoxide anion and ferrous cation [10]:

[MATHEMATICAL EXPRESSION NOT REPRODUCIBLE IN ASCII]

In the reaction of [O.sub.2.sup.*-] and [H.sub.2][O.sub.2], singlet oxygen ([sup.1][O.sub.2]), hydroxide radical (O[H.sup.*]) and hydroxide anion (O[H.sup.-]) are formed

[O.sub.2.sup.*-] + [H.sub.2][O.sub.2] [right arrow] [sup.1][O.sub.2] + O[H.sup.*] + O[H.sup.-]

Reactive nitrogen species

Reactive nitrogen species, such as nitrogen oxide radical (N[O.sup.*]), nitrogen dioxide radical (N[O.sub.2.sup.*]), nitrosonic cation (N[O.sup.+]), nitroxyl(ic) anion (N[O.sup.-]) and peroxinitrite (ONO[O.sup.-]) are present in erythrocytes [11]. Nitrogen oxide is synthesized in the cells of the vascular endothelium, from where it is transported to the cells of vascular smooth muscles and to the plasma [12]. From the plasma it passes to erythrocytes, where it is bounded to hemoglobin. The reaction of binding nitrogen oxide to hemoglobin comprises S-nitrolisation of Cys93 residue in the beta-globin chain and formation of S-nitrosohemoglobin (HbSNO) [13]. The reaction takes place with oxygenated hemoglobin in R conformation [14]. Deoxygenation of hemoglobin and its passage to T conformation releases N[O.sup.*] back to the plasma. Due to these processes, venous blood contains approximately 10fold less HbSNO than arterial blood with S-nitrosilohemoglobin being an N[O.sup.*] transporter. Nitrogen oxide is transported from HbSNO, in the S-transnitrosilation, to fine particulate thioles such as cysteine and glutathione with the formation of nitrosocysteine (CysNO) and nitrosoglutathione (GSNO) [15]. S-nitrosilation is easier while denitrosilation is harder in case of glycated hemoglobin [16]. S-nitrosilation of pathological hemoglobin S (HbS) hinders its polymerization. Increased content of S-nitrosohemoglobin can be observed in endotoxemia.

In the condition of oxidative stress, the reaction of nitrogen oxide and oxyhemoglobin ferrous cation with superoxide anion, takes place. The reaction of nitrogen oxide and oxyhemoglobin iron leads to the formation of methemoglobin and nitrates [17]:

Hb[O.sub.2] + N[O.sup.*] [right arrow] MetHb + N[O.sub.3.sup.-]

Peroxinitrite anion is the product of the reaction of nitrogen oxide and superoxide anion [18]:

[O.sub.2.sup.*-] + N[O.sup.*] [right arrow] ONO[O.sup.-]

Spontaneous protonation of the peroxynitrite leads to the formation of nitrogen dioxide (N[O.sub.2]) and hydroxyl radical (O[H.sup.*]):

ONO[O.sup.-] + [H.sup.+] [right arrow] N[O.sub.2] + O[H.sup.*]

In the reaction of peroxynitrite with sulfhydric compounds, their nitro-derivatives are formed [19]:

ONO[O.sup.-] + RSH [right arrow] RSNO + [O.sub.2] + [H.sup.+]

Reactions mentioned above make the circulating blood poorer in nitrogen oxide.

Deficiency of nitrogen oxide and its highly reactive metabolic products specifically peroxinitrite, cause the membrane proteins damage and decrease erythrocytes flexibility [20, 21]. The metabolism of reactive nitrogen species in erythrocyte is presented in Fig. 2 [19].

Antioxidants

Both reactive oxygen species and reactive nitrogen species that are formed in erythrocytes are inactivated by antioxidative enzymes and non-enzymatic antioxidants.

Due to the antioxidants activity, the amount of methemoglobin in erythrocytes does not exceed 0.6-1.8% of hemoglobin in erythrocytes. There are three kinds of reductases that reduce trivalent iron of methemoglobin, NADH is the coenzyme of the first one, NADPH--the second one, and the third one acts with participation of ferrocytochrome [b.sub.5] [22].

Reductase of methemoglobin: NADH catalyses the reaction of:

MetHb-[Fe.sup.3+] + NADH [right arrow] Hb-[Fe.sup.2+] + NAD

The course of reaction catalyzed by reductase of methemoglobin: NADPH goes as follows:

MetHb-[Fe.sup.3+] + NADPH [right arrow] Hb-[Fe.sup.2+] + NADP

The reduction of methemoglobin by ferrocytochrome [b.sub.5] follows the scheme:

MetHb-[Fe.sup.3+] + ferrocytochrom [b.sub.5] [right arrow] Hb-[Fe.sup.2+] + ferrocytochrom [b.sub.5]

Cytozylic CuZn-dismutase superoxide catalyses the reaction of dismutation of superoxide anion to less reactive hydrogen peroxide and molecular oxygen [23]:

2[O.sub.2.sup.*-] + 2[H.sup.+] [right arrow] [H.sub.2][O.sub.2] + [O.sub.2]

Catalase decomposes approximately 50% of formed hydrogen peroxide in erythrocytes [24]:

2[H.sub.2][O.sub.2] [right arrow] 2[H.sub.2]O + [O.sub.2]

Hemoglobin, comprising about 10% of catalase activity of erythrocyte, decomposes hydrogen peroxide in the non-enzymatic reaction at the lower rate than catalase herself does [25]. Due to high content of hemoglobin, its antioxidative activity has a great impact on the regulation of erythrocyte oxidative-antioxidative processes.

Glutathione thiolic groups [26] and protein thiolic groups [27] have an important antioxidative role in the erythrocyte and comprise approximately

0.1% of total mass of these cells with reduced glutathione (GSH) of 99.8% and oxidized glutathione (GSSG)--0.2% of its content. Oxidized glutathione also occurs in combination with thiolic groups of hemoglobin and other proteins of the erythrocyte, in the form of mixed disulphides (PSSG). GSSG and PSSG contents in erythrocytes increase in the oxidative stress [28].

Reduced glutathione decomposes hydrogen peroxide directly and in the reaction catalyzed by selenium-dependent glutathione peroxidase [26]:

[H.sub.2][O.sub.2] + 2GSH [right arrow] 2[H.sub.2]O + GSSG

GSH also participates in the reaction of decomposition of alcylhydroxides (ROOH) to alcohol (ROH) [29]:

ROOH + 2GSH [right arrow] ROH + [H.sub.2]O + GSSG

Moreover, GSH is coupled with hydroxyl derivatives of xenobiotics (X-OH) in the reaction catalyzed by S-transferase of glutathione [29]:

X-OH + GSH [right arrow] X-SG + [H.sub.2]O

Reductase oxidized glutathione: NADPH reduces oxidized glutathione:

GSSG + NADPH [right arrow] 2GSH + NADP

Ascorbate, cysteine and ergothioneine are fine particulate hydrophilic antioxidants of the erythrocyte, localized in cytosol [26]. Alpha-tocopherol [30] and beta-carotene [31] are fine particulate hydrophobic antioxidants, localized in the erythrocyte membrane. Ascorbate, alpha-tocopherol, beta-carotene and ergothionein are exogenic antioxidants.

The reaction of reactive nitrogen species and thiolic groups, with formation of S-nitrozotioli (RSNO), leads to their detoxication [32].

Oxidative-antioxidative balance

Reactive oxygen species and reactive nitrogen species are natural products of erythrocyte metabolism and are in a dynamic balance with enzymatic and non-enzymatic antioxidants. Inhibition of dismutase activity by hydrogen superoxide and inhibition of catalase activity by superoxide anion prove co-dependence of these systems [33].

Singlet oxygen and hydroxyl radical are the most reactive forms of oxygen while peroxinitrite--the most reactive form of nitrogen. They are characterized by low specificity and react with almost every organic molecule: they attach or separate an electron to or from the molecule and form an appropriate free radical.

The activity of oxidative and antioxidative factors of the blood plasma has an effect on oxidative-antioxidative balance of the erythrocytes. The activity of erythrocyte antioxidative enzymes is significantly higher than that of blood plasma (Tab. 2) [34].

The content of fine particulate antioxidants is also significantly higher in erythrocytes than in blood plasma [35]. Thus, internal surface of erythrocyte membranes is better protected by reactive oxygen species than the external surface. A significant antioxidative role is fulfilled by the binding of hemoglobin through haptoglobin [36] and ferric cations through transferrin [37].

Erythrocyte aging, apoptosis, hemoglobin degradation

Elimination of cells of the red-blood-cell system, at each stage of their development, occurs in the course of apoptosis [38]. Causative and executive factors as well as the range and course of apoptosis are different in different developmental phases of the erythrocyte. As far as erythroblasts are concerned, total apoptosis concerns only damaged, mutated, or infected cells. The process of apoptosis is initiated by typical apoptotic factors and performed by typical apoptosis executors leading to mitochondria, cellular nucleus and the whole cell damage. Erythropoietin protects non-damaged erythroblasts against total apoptosis [39]. There are only limited apoptotic processes in non-damaged erythroblasts which concern cellular organelles and lead to their fragmentation and decomposition. The processes of limited apoptosis are continued in reticulocytes. Total decomposition of ribosomes and mitochondria occurs and their fragments are eliminated in form of coated vesicles.

Mature erythrocytes without cellular organelles are metabolically stable. Only old and used up erythrocytes, which life span in blood reached about 120 days, undergo apoptosis and elimination from the bloodstream. In 24 hours, 0,21 x [10.sup.12] erythrocytes (0.83% of total) are eliminated from blood and about 8g of hemoglobin (1% of total) is degraded [40]. Deferoxamine protects against elimination processes [41]. In pathological conditions, e.g. in hemolytic anemia and in intoxication with certain xenobiotics, life span of erythrocytes in circulating blood is shortened and they undergo apoptosis earlier.

In aging erythrocytes, apoptosis is induced by appearance of increased amount of reactive oxygen species [42] and increased content of calcium cations [43]. The calcium cations activate procaspase-3 and procaspase-8 [2] while the procaspase-8 activate calpain I ([mu]-calpain) [44]. The activity of above enzymes is inhibited by specific inhibitors [45]. In aging occur changes in erythrocytes characteristic for apoptosis, such as decrease in affinity to hemoglobin, decrease in activity of enzymes of glycolytic cycle and pentose cycle, oxidation and fragmentation of proteins. Apoptotic changes in erythrocytes concern also: the chemical composition (decrease in content of glutathione and calcium cations and increase in content of methemoglobin, oxidized lipids and sodium cation), the structure of erythrocyte membrane (translocation of phosphatidylserine from the internal to the external monolayer, loss of peripheral membrane proteins, disturbance of lipid asymmetry) and morphological features (change of shape from biconcave to spherical, decrease in size through shrinking, changes of surface from smooth to that with bulge. Translocation of phosphatidylserine to the external layer of the membrane and forming bulges on its surface is the most important change [46]. Above described changes in external layer of the red cell membrane begin the process of apoptosis and enables recognition as well as binding of erythrocyte by receptors of mononuclear phagocytes of sinusoid vessels of the spleen where decomposition of apoptotic erythrocytes total occur.

In used up erythrocytes, oxidative modification of the residues of certain amino-acids takes place as well as disturbs of the spatial structure of hemoglobin and membrane proteins, which increases their susceptibility to proteases activities [47]. Oxidatively modified hemoglobin is degraded by multi-catalytic proteasomal system [48,49] and modified membrane proteins--by membrane proteases [50]. Fragmentation of hemoglobin and membrane proteins is observed, with end products of fine particulate peptides and amino-acids, resulting in erythrocyte decomposition. Heme, with its oxidative activity, is released in erythrocyte decomposition processes. Heme oxygenase decomposes heme to biliverdin, ferrous cation and carbon monoxide [51]. Biliverdin is oxidized to bilirubin, a highly antioxidative compound [52].

In erythrocytes with metabolic defects, containing pathological hemoglobin, increased methemoglobin content, with deficiency of heme oxidase and enzymes reducing oxidized glutathione, deep changes in protein structure and morphological changes take place [53, 54], which is presented in Figure 3. In Figure 3 arrow number 1 indicates factors modifying HB--congenital molecular defects and those caused by reactive oxygen species; arrow number 2--indicates factors increasing the formation of reactive oxygen species: increased oxygen supply, ionizing radiation, ultraviolet, and heat radiation, ultrasounds, xenobiotics, certain medicines; arrow number 3 indicates factors decreasing reactive oxygen species decomposition: enzymes deficiency (glucose-6-phosphate dehydrogenase, glutathione peroxidase, glutathione reductase and synthetase, catalase, superoxide dismutase), fine particulate compound deficiency (alpha-tocopherol, ascorbate, beta-carotene, glutathione). The oxidation of thiolic groups of hemoglobin leads to the formation of disulphuric bindings inside subunits and between subunits of hemoglobin, which leads to the formation of intramolecular cross-binding and intermolecular bindings as well as the formation of insoluble aggregates [55]. Proteins modified in above described way lose their ability to solubility and are resistant to protease activity [56]. Protein losses and bodies composed of denaturated hemoglobin and proteins of erythrocyte stroma, called Heinz bodies, occur [57,58]. On the other hand, uncontrolled oxidative and proteolytic fragmentation of erythrocyte membrane proteins leads to their hemolysis [9].

Erythrocytes in blood extravasated to tissues also undergo apoptosis. The inflammatory process and necrosis appear only when apoptotic changes are accompanied by bacterial infection and there occurs leukocyte infiltration.

Methodological points

Methods of identification and quantitative determination of reactive forms of oxygen and nitrogen, activity of enzymes participating in their formation, contents of products of activities of reactive oxygen species and reactive nitrogen species to lipids, proteins and nucleic acids as well as activity of antioxidative enzymes and concentration of non-enzymatic antioxidants are described by many authors [59, 60].

Determination of activity of most components in erythrocytes does not require modification although hemoglobin can interfere with determination of certain components. Superoxidative dismutase can be determined only after prior removal of hemoglobin using the mixture of chloroform and ethanol [61]. Thiolic compounds are determined after the reaction with 4,4'-ditiodipirydyne and absorbance can be measured at 324 nm, which makes the result hemoglobin-independent [62]. Cysteine is determined using ninhydrinic agent in strongly acidic environment [63]. Ergothioneine is determined using spectrophotometric method, after the reaction with disulfide 2,2'-dipyrydole [64].

The evaluation of components of the erythrocyte oxidative-antioxidative system should be performed in the most recently collected blood samples and isolated erythrocytes.

The results are counted per 1ml of packed erythrocytes, containing [10.sup.10] of these cells. Erythrocyte life span, osmotic resistance, ability to change size, shape, stainability and occurrence of intraerythrocyte aposomes are used to evaluate the effects of activity of reactive oxygen species to erythrocytes [65,66].

Acknowledgments

The authors thank Professor Krzysztof Worowski for help in editing this article,

Conflicts of interest

Actual or potential conflicts of interest do not exist.

REFERENCES

[1.] Rapoport SM, Schewe T. The maturational breakdown of mitochondria in reticulocytes. Biochim Biophys Acta. 1986 Dec 22;864(3-4):471-95.

[2.] Frendo J. Funkcje biochemiczne komorek krwi i szpiku. W: Fizjologia krwi. 1st ed. Warszawa: PWN; 1998. Dqbrowski Z; p. 114-42.

[3.] Berg CP, Engels IH, Rothbart A, Lauber K, Renz A, Schlosser SF, Schulze-Osthoff K, Wesselborg S. Human mature red blood cells express caspase-3 and caspase-8, but are devoid of mitochondrial regulators of apoptosis. Cell Death Differ. 2001 Dec;8(12):1197-206.

[4.] Misztal T, Tomasiak M. Pathophysiological consequences of hemolysis. Role of cell-free hemoglobin. Post Hig Med Dosw. 2011 Sep 28;65:627-39.

[5.] Bynoe LA, Gottsch JD, Pou S, Rosen GM. Light-dependent generation of superoxide from human erythrocytes. Photochem Photobiol. 1992 Sep;56(3):353-6.

[6.] Winterbourn CC. Oxidative denaturation in congenital hemolytic anemias: the unstable hemoglobins. Semin Hematol. 1990 Jan; 27(1):41-50.

[7.] De Bono DP. Free radicals and antioxidants in vascular biology: the roles of reaction kinetics, environment and substrate turnover. QJM. 1994 Aug; 87(8):445-53.

[8.] Sadrzadeh SM. Graf E, Panter SS. Hallaway PE, Eaton JW. Hemoglobin. A biologic Fenton reagent. J Biol Chem. 1984 Dec10;259 (23):14354-6.

[9.] Carrell RW, Winterbourn CC, Rachmilewitz EA. Activated oxygen and haemolysis. Br J Haematol. 1975 Jul; 30(3):259-64.

[10.] Kehrer JP. The Haber-Weiss reaction and mechanisms of toxicity. Toxicology. 2000 Aug 14;149(1):43-50.

[11.] Pryor WA, Squadrito GL. The chemistry of peroxynitrite: a product from the reaction of nitric oxide with superoxide. Am J Physiol. 1995

May; 268(5 Pt 1):L699-722.

[12.] Gladwin MT, Lancaster JR, Freeman BA, Schechter AN. Nitric oxide's reactions with hemoglobin: a view through the SNO-storm. Nat Med. 2003 May;9(5):496-500.

[13.] Gladwin MT, Ognibene FP, Pannell LK, Nichols JS, Pease-Fye ME, Shelhamer JH, Schechter AN. Relative role of heme nitrosylation and beta-cysteine 93 nitrosation in the transport and metabolism of nitric oxide by hemoglobin in the human circulation. Proc Natl Acad Sci USA. 2000 Aug 29; 97(18):9943-8.

[14.] Huang Z, Shiva S, Kim-Shapiro DB, Patel RP, Ringwood LA, Irby CE, Huang KT, Ho C, Hogg N, Schechter AN, Gladwin MT. Enzymatic function of hemoglobin as a nitrite reductase that produces NO under allosteric control. J Clin Invest. 2005 Aug;115(8):2099-107.

[15.] Sandmann J, Schwedhelm KS, Tsikas D. Specific transport of S-nitrosocysteine in human red blood cells: Implications for formation of S-nitrosothiols and transport of NO bioactivity within the vasculature. FEBS Lett. 2005 Aug 1; 579(19):4119-24.

[16.] Panzer S, Kronik G, Lechner K, Bettelheim P, Neumann E, Dudczak R. Glycosylated hemoglobins (GHb): an index of red cell survival. Blood. 1982 Jan; 59(6):1348-50.

[17.] Bouton C. Nitrosative and oxidative modulation of iron regulatory proteins. Cell Mol Life Sci. 1999 Jul;55(8-9):1043-53.

[18.] Radi R, Beckman JS, Bush KM, Freeman BA. Peroxynitrite oxidation of sulfhydryls. The cytotoxic potential of superoxide and nitric oxide. J Biol Chem. 1991 Mar 5;266(7):4244-50.

[19.] Loscalzo J. Nitric oxide insufficiency, platelet activation, and arterial thrombosis. Circ Res. 2001 Apr 27;88(8):756-62.

[20.] Boccini F, Herold S. Mechanistic studies of the oxidation of oxyhemoglobin by peroxynitrite. Biochemistry. 2004 Dec 28; 43(51):16393-404.

[21.] Radi R, Beckman JS, Bush KM, Freeman BA. Peroxynitrite-induced membrane lipid peroxidation: the cytotoxic potential of superoxide and nitric oxide. Arch. Biochem. Biophys. 1991 Aug 1;288(2):481-7.

[22.] Hultquist DE, Xu F, Quandt KS, Shlafer M, Mack CP, Till GO, Seekamp A, Betz AL, Ennis SR. Evidence that NADPH-dependent methemoglobin reductase and administered riboflavin protect tissues from oxidative injury. Am J Hematol. 1993 Jan;42(1):13-8.

[23.] Concetti A, Massei P, Rotilio G, Brunori M, Rachmilewitz EA. Superoxide dismutase in red blood cells: method of assay and enzyme content in normal subjects and in patients with P-thalassemia (major and intermedia). J Lab Clin Med. 1976 Jun; 87(6):1057-64.

[24.] Gaetani GF, Ferraris AM, Rolfo M, Mangerini R, Arena S, Kirkman HN. Predominant role of catalase in the disposal of hydrogen peroxide within human erythrocytes. Blood. 1996 Feb 15; 87(4):1595-9.

[25.] Gabbianelli R, Santroni AM, Fedeli D, Kantar A, Falcioni G. Antioxidant activities of different hemoglobin derivatives. Biochem Biophys Res Commun. 1998 Jan 26; 242 (3):560-4.

[26.] Wlodek L. Glutation. In: Biotiole. Krakow: UJ; 2003. p. 111-60.

[27.] Di Simplicio P, Cacace MG, Lusini L, Giannerini F, Giustarini D, Rossi R. Role of protein-SH groups in redox homeostasis the erythrocyte as a model system. Arch Biochem Biophys. 1998 Jul 15;355(2):145-52.

[28.] Murakami K, Mawatari S. Oxidation of hemoglobin to methemoglobin in intact erythrocyte by a hydroperoxide induces formation of glutathionyl hemoglobin and binding of a-hemoglobin to membrane. Arch Biochem Biophys. 2003 Sep 15;417(2):244-50.

[29.] Sies H. Strategies of antioxidant defense. Eur J Biochem. 1993 Jul 15;215(2):213-9.

[30.] Tanaka H, Mino M. Membrane-to-membrane transfer of tocopherol in red blood cells. J Nutr Sci Vitaminol. 1986 Oct; 32(5):463-74.

[31.] Paiva SA, Russell RM. Beta-carotene and other carotenoides as antioxidants. J Am Coll Nutr. 1999 Oct;18(5):426-33.

[32.] Hughes MN. Relationship between nitric oxide, nitroxyl ion, nitrosonium cation and peroxynitrite. Biochim Biophys Acta. 1999 May 5; 1411(2-3):263-72.

[33.] Hodgson EK, Fridovich I. The interaction of bovine erythrocyte superoxide dismutase with hydrogen peroxide: inactivation of the enzyme. Biochemistry. 1975 Dec 2; 14(24):5294-9.

[34.] Frei B, Stocker R, Ames BN. Antioxidant defenses and lipid peroxidation in human blood plasma. Proc Natl Acad Sci USA. 1988 Dec; 85(24):9748-52.

[35.] Sies H, de Groot H. Role of reactive oxygen species in cell toxicity. Toxicol Lett. 1992 Dec; 64-65 Spec:547-51.

[36.] Wassell J. Haptoglobin: function and polymorphism. Clin Lab. 2000; 46(11-12):547-52.

[37.] Agarwal R. Transferrin saturation with intravenous irons: an in vitro study. Kidney Int. 2004 Sep; 66(3):1139-44.

[38.] Delaunay J. Molecular basis of red cell membrane disorders. Acta Haematol. 2002;108(4):210-8.

[39.] Silva M, Grillot D, Benito A, Richard C, Numez G, Fernandez-Luna JL. Erythropoietin can promote erythroid progenitor survival by represing apoptosis. Blood. 1996 Sep 1; 88(5):1576-82.

[40.] Uchida I, Tashiro C, Koo YH, Mashimo T, Yoshiya I. Carboxyhemoglobin and methemoglobin levels in banked blood. J Clin Anesth. 1990 Mar-Apr;2(2):86-90.

[41.] Knight JA, Searles DA, Clayton FC. The effect of desferrioxamine on stored erythrocytes: lipid peroxidation, deformability, and morphology. Ann Clin Lab Sci. 1996 Jul-Aug; 26(4):283-90.

[42.] Di Cola D, Sacchetta P, Battista P. Proteolysis in human erythrocytes is triggered only by selected oxidative stressing agents. Ital J Biochem. 1988 May-Jun;37(3):129-38.

[43.] Bratosin D, Estaquier J, Ameisen JC, Montreuil J. Molecular and cellular mechanisms of erythrocyte programmed cell death: impact on blood transfusion. Vox Sang. 2002 Aug; 83 Suppl 1:307-10.

[44.] Glaser T, Schwarz-Benmeir N, Barnoy S, Barak S, Eshhar Z, Kosower NS. Calpain ([Ca.sup.2+]-dependent thiol protease) in erythrocytes of young and old individuals. Proc Natl Acad Sci USA. 1994 Aug 16; 91(17):7879-83.

[45.] Schwarz-Benmeir N, Glaser T, Barnoy S, Kosower NS. Calpastatin in erythrocytes of young and old individuals. Biochem J. 1994 Dec 1;304(Pt 2):365-70.

[46.] Closse C, Dachary-Prigent J, Boisseau MR. Phosphatidylserine-related adhesion of human erythrocytes to vascular endothelium. Br J Haematol. 1999 Nov; 107(2):300-2.

[47.] Percy MJ, McFerran NV, Lappin TRJ. Disorders of oxidised haemoglobin. Blood Rev. 2005 Mar;19(2):61-8.

[48.] Giulivi C, Pacifici RE, Davies KJ. Exposure of hydrophobic moieties promotes the selective degradation of hydrogen peroxide-modified hemoglobin by the multicatalytic proteinase complex, proteasome. Arch Biochem Biophys. 1994 Jun;311(2):329-41.

[49.] Pacifici RE, Kono Y, Davies K.J. Hydrophobicity as the signal for selective degradation of hydroxyl radical-modified hemoglobin by the multicatalytic proteinase complex, proteasome. J Biol Chem. 1993 Jul 25;268(21):1505-11.

[50.] Beppu M, Inoue M, Ishikawa T, Kikugawa K. Presence of membrane-bound proteinases that preferentially degrade oxidatively damaged erythrocyte membrane proteins as secondary antioxidant defense. Biochim Biophys Acta. 1994 Nov 23;1196(1):81-7.

[51.] Deshane J, Wright M, Agarwal A. Heme oxygenase-1 expression in disease states. Acta Biochim Pol. 2005;52(2):273-84.

[52.] Tomaro ML, Batlle AM. Bilirubin: its role in cytoprotection against oxidative stress. Int J Biochem Cell Biol. 2002 Mar;34(3):216-20.

[53.] Baskurt OK, Temiz A, Meiselman HJ. Effect of superoxide anions on red blood cell rheologic properties. Free Radic Biol Med. 1998 Jan;24(1):102-10.

[54.] Tavazzi B, Di Pierro D, Amorini AM, Fazzina G, Tuttobene M, Giardina B, Lazzarino G. Energy metabolism and lipid peroxidation of human erythrocytes as a function of increased oxidative stress. Eur J Biochem. 2000 Feb; 267 (3):684-9.

[55.] Thillet J, Michelson AM. Oxidation and cross-linking of human hemoglobins by activated oxygen species. Free Radic Res Commun. 1985;1(2):89-100.

[56.] Chiu DT, Liu TZ. Free radical and oxidative damage in human blood cells. J Biomed Sci. 1997;4(5):256-9.

[57.] Chiu D, Lubin B. Oxidative hemoglobin denaturation and RBC destruction: the effect of heme on red cell membranes. Semin Hematol. 1989 Apr;26(2):128-35.

[58.] Platt OS, Falcone JF. Membrane protein lesions in erythrocytes with Heinz bodies. J Clin Invest. 1988 Sep;82(3):1051-8.

[59.] Antolovich M, Prenzler PD, Patsalides E, McDonald S, Robards K. Methods for testing antioxidant activity. Analyst. 2002 Jan;127 (1):183-98.

[60.] Rice-Evans CA, Diplock AT, Symons MCR. Techniques in free radical research. Amsterdam: Elsevier; 1991.

[61.] Fu RH, Yang PH, Chiang MC, Chiang CC, Cho YH, Chou YH. Erythrocyte Cu/Zn superoxide dismutase activity in preterm infants with and without bronchopulmonary dysplasia. Biol Neonate. 2005; 88(1):35-41.

[62.] Grassetti DR, Murray JF. Determination of sulfhydryl groups with 2,2'- or 4,4'-dithiodipyridine. Arch Biochem Biophys. 1967 Mar;119(1):41-9.

[63.] Gaitonde MK. A spectrophotometric method for the direct determination of cysteine in the presence of other naturally occurring amino acids. Biochem J. 1967 Aug;104(2):627-33.

[64.] Mitsuyama H, May JM. Uptake and antioxidant effects of ergothioneine in human erythrocytes. Clin Sci. 1999 Oct; 97(4):407-11.

[65.] Bocci V. Determinants of erythrocyte ageing: a reapraisal. Br J Haematol. 1981 Aug; 48(4): 515-22.

[66.] Shiga T, Maeda N, Kon K. Erythrocyte rheology. Crit Rev Oncol Hematol. 1990; 10 (1):9-48.

Lapinski R. *, Siergiejuk M., Worowska A., Gacko M.

Department of Vascular Surgery and Transplantology, Medical University of Bialystok, Poland

* Corresponding author:

Department of Vascular Surgery and Transplantology Medical University of Bialystok 24A Sklodowskiej-Curie Str. 15-276 Bialystok, Poland

Tel.: +48 85 746 82 77; Fax: +48 85 746 88 96

e-mail: chirnacz@umwb.edu.pl

Received: 13.01.2014

Accepted: 26.05.2014

Table 1. Chemical composition of erythrocyte [5] *

Component                   Contents, %

                          total   dry mass

water                     65.0      0.0
remaining compounds       35.0     100.0
hemoglobin                 33      94.00
non-hemoglobin proteins    ~ 1     ~ 2.80
lipids                     0.5      1.40
heme iron                 0.11      0.32
glutathione               ~ 0.1    ~ 0.28
peroxide dismutase        0.007    0.019
other compounds           ~ 0.4    ~ 1.14

*--mean approximate values

Table 2. Activity of antioxidative enzymes of
erythrocytes and blood plasma [34].

                     Activity

Enzyme         Erythrocytes      Plasma
              U/[10.sup.10] *     U/ml

Superoxide        550-800         5-20
dismutase
Catalase         3800-5400       small
                                quantity
Glutathione      7.8-10.6         0.4
peroxidase
Glutathione       2.7-3.7         0.03
reductase
Glutathione       1.5-2.5        0.005
transferase

*--number of erythrocytes contained in 1 ml.
COPYRIGHT 2014 Medical University of Bialystok
No portion of this article can be reproduced without the express written permission from the copyright holder.
Copyright 2014 Gale, Cengage Learning. All rights reserved.

Article Details
Printer friendly Cite/link Email Feedback
Author:Lapinski, R.; Siergiejuk, M.; Worowska, A.; Gacko, M.
Publication:Progress in Health Sciences
Date:Jun 1, 2014
Words:5116
Previous Article:Extrajudicial ways of compensating for medical damage in Poland.
Next Article:Ageing society--a review of strategies for action.
Topics:

Terms of use | Privacy policy | Copyright © 2021 Farlex, Inc. | Feedback | For webmasters |