Erythrocytes peroxide hemolysis

Clinical manifestations of vitamin E deficiency may be seen in premature infants fed on formnlas high in polyunsaturated fatty acids (H7). In a group of 6 infants on a nonfat powdered cow s milk formula with added cottonseed oil plasma vitamin E levels decreased from 0.22 0.04 (SE) mg/100 ml at birth to 0.08 0.03 mg/100 ml at 4 weeks. Inversely, the in vitro erythrocyte sensitivity to peroxide hemolysis increased from 18 4 (SE)% at birth to 77 14 (SE)% at 4 to 5 weeks after birth. In general, the clinical symptoms were mild, consisting of edema and skin lesions which appeared at about 4 weeks of age. 

The. animal work will be described only sufficiently to allow the reader to appreciate lietter the discussion of the human studies. When it became apparent in 19.55 that the peroxide hemolysis test was dependent not only on the tocopherol level of the blood but also upon the level of linoleic acid (and other autoxidizable components) in the stroma of the erythrocyte, animal experiments were designed to obtain more exact correlations between tocopherol needs and linoleic acid intake. This relationship between linoleic acid content of the diet and the incidence of chick encephalomalacia (Century and Horwitt, 19.58) was not recorded until later (Century et al., 19.59 Century and Horwitt, 19.59) when observation of cerebellar encephalomalacia in an infant that had been fed a commercial cottonseed oil preparation intravenously came to our attention (Horwitt and Bailey, 1959). In the meantime, there had been a number of reports to certify the relationship between linoleic acid consumption and chick encephalomalacia (Dam et al, 19.58 Machlin and Gordon, 1960). With the advent of better gas chromatographic techniques, it was soon possible to show that the linoleic acid content of the cerebellum was diet dependent (Horwitt et al., 1959 Witting et al., 1961). The marked effects of diet on the fatty acids of the mitochondria of chick brains has also been reported (Horwitt, 1981a). The levels of linoleic acid are much lower in brain tissues than in any tissue analyzed to date and this relatively low linoleic acid level may be considered a characteristic of brain tissue. The significance of this difference is not known. It is of interest to note that the current interpretations of the effect of more unsaturated fats on the production of chick encephalomalacia were anticipated by Dam in 1944. 

In order to estimate whether there is any possible relationship of the in vitro peroxide hemolysis test with the survival rate of the erythrocytes... 

In 1977, Kellogg and Fridovich [28] showed that superoxide produced by the XO-acetaldehyde system initiated the oxidation of liposomes and hemolysis of erythrocytes. Lipid peroxidation was inhibited by SOD and catalase but not the hydroxyl radical scavenger mannitol. Gutteridge et al. [29] showed that the superoxide-generating system (aldehyde-XO) oxidized lipid micelles and decomposed deoxyribose. Superoxide and iron ions are apparently involved in the NADPH-dependent lipid peroxidation in human placental mitochondria [30], Ohyashiki and Nunomura [31] have found that the ferric ion-dependent lipid peroxidation of phospholipid liposomes was enhanced under acidic conditions (from pH 7.4 to 5.5). This reaction was inhibited by SOD, catalase, and hydroxyl radical scavengers. Ohyashiki and Nunomura suggested that superoxide, hydrogen peroxide, and hydroxyl radicals participate in the initiation of liposome oxidation. It has also been shown [32] that SOD inhibited the chain oxidation of methyl linoleate (but not methyl oleate) in phosphate buffer. 

Quercetin and rutin suppressed photosensitized hemolysis of human erythrocytes with ho values equal to 40 p.mol l-1 and 150 jjlmt>I I 1, respectively [139]. Suppression of photohemolysis was accompanied by inhibition of lipid peroxidation. Morin inhibited oxygen radical-mediated damage induced by superoxide or peroxyl radicals to the human cells in the cardiovascular system, erythrocytes, ventricular myocytes, and saphenous vein endothelial cells [140]. Rutin protected against hemoglobin oxidation inside erythrocytes stimulated by prooxidant primaquine [141],... 

In 1989, we showed [142] that the Fe2+(rutin)2 complex is a more effective inhibitor than rutin of asbestos-induced erythrocyte hemolysis and asbestos-stimulated oxygen radical production by rat peritoneal macrophages. Later on, to evaluate the mechanisms of antioxidant activities of iron rutin and copper-rutin complexes, we compared the effects of these complexes on iron-dependent liposomal and microsomal lipid peroxidation [165], It was found that the iron rutin complex was by two to three times a more efficient inhibitor of liposomal peroxidation than the copper-rutin complex, while the opposite tendency was observed in NADPH-dependent microsomal peroxidation. On the other hand, the copper rutin complex was much more effective than the iron rutin complex in the suppression of microsomal superoxide production, indicating that the copper rutin complex indeed acquired additional SOD-dismuting activity because superoxide is an initiator of NADPH-dependent... 

It has been known since the early 1950s that turmeric had strong antioxidant effects with curcumin being the major compound responsible followed by demethoxycurcumin and bisdemethoxycurcumin. All three inhibit lipid peroxidation and have a positive anti-oxidant effect for hemolysis and lipid peroxidation in mouse erythrocytes.11 Curry pills containing turmeric are being marketed as a prevention for colon cancer.29... 

The accumulation of hydrogen peroxidase affects many intracellular processes and results in hemolysis. These include the cross-linking of membrane proteins hemoglobin denaturation (manifest as Heinz body formation), which in turn affects the physical properties of the erythrocyte and lipid peroxidation, which may affect the cell membrane to cause direct hemolysis (Fig. 11-8). The resultant damage leads to a mixture of intravascular hemolysis and extravascu-lar hemolysis (by which hemolysis occurs in the reticuloendothelial system). In acute hemolytic episodes, the clinical picture is of predominantly intravascular hemolysis, while predominantly extravascular hemolysis is seen in patients with chronic hemolysis. 

Valenzuela, A. Guerra, R. Garrido, A. Silybin dihemisuc-cinate protects rat erythrocytes against phenylhydrazine-induced lipid peroxidation and hemolysis. Planta Med. 1987, 53, 402-A05. 

It is difficult to produce vitamin E deficiency in adult humans. Adult males who were depleted of vitamin E for 6 years showed no symptoms, although serum tocopherol concentrations became very low. However, their erythrocytes lysed more readily than normal when exposed to hydrogen peroxide or other oxidizing agents in vitro. This finding led to the use of low-plasma vitamin E and increased susceptibility of erythrocytes to oxidative hemolysis as criteria for vitamin E deficiency. 

Pyridoxatin was 20 times as active (IC50 = 0.55 pg/ml) as vitamin E as an inhibitor of free radical induced lipid peroxidation in rat liver microsomes [221]. It inhibited (IC50= 1-95 Ug/ml) hemolysis of rat erythrocytes catalyzed by a free radical initiator [221]. Pyridoxatin inhibited the growth of HeLa cells (IC50 = 1.0 pg/ml), but only displayed antimicrobial activity against Candida albicans (MIC = 1.64 pg/ml) [221]. 

Using Cr -labeled cells, a shortened erythrocyte life span was found in newborn premature infants (Kaplan and Hsu, 1961). The susceptibility of their erythrocytes to in vitro hemolysis in hydrogen peroxide suggests a possible relationship to their decreased survival in vivo. Indeed, decreased survival has been observed in the vitamin E-deficient monkey (Marvin et al., 1959) although the normal survival in vitamin E-defident rats suggests a species variability (Goldbloom, 1960). The results in humans are inconclusive. A decreased red cell life span was found in adult volunteers on a tocopherol-deficient diet high in polyunsaturated lipids (Horwitt et al., 1961), but survival was normal in a patient with cystic fibrosis of... 

We are thus forced to make an estimate concerning the need for tocopherol supplementation from inadequate data. In the case of the full-term infant, there would seem to be no need for supplementation of the diet, since on ordinary cows milk mixtures within 6 months the level of tocopherol is above that at which hemolysis of erythrocytes occurs in hydrogen peroxide in vitro. This level is reached in the first week by babies fed breast... 

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