Effects of Ascorbic Acid on Peroxidation of Human Erythrocyte Membranes by Lipoxygenase.

Shiro, Motoo; Murakami, Kaori

doi:10.3177/jnsv.45.687

Cited by 3 publications

(6 citation statements)

References 35 publications

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“…The HPLC method for separation of the erythrocyte membrane PL was modified somewhat from our previous report (28,29). The column was a Wakosil 5 NH 2 , 150 × 4.6 mm (Wako Pure Chemical Co.) and the guard column was a µ-Bondpak NH 2 , 20 × 4 mm (Waters Co.).…”

Section: Methodsmentioning

confidence: 99%

“…We recently reported a method employing high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection that can detect all of the major PL classes of erythrocyte membranes including phosphatidylserine (PS) by means of a single chromatographic elution (28,29). We applied the HPLC method to investigate changes of membrane PL in intact human erythrocytes; these changes are expressed as the ratio of each PL to sphingomyelin (SM).…”

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confidence: 99%

“…We applied the HPLC method to investigate changes of membrane PL in intact human erythrocytes; these changes are expressed as the ratio of each PL to sphingomyelin (SM). Furthermore, α-Toc, γ-tocopherol (γ-Toc), and cholesterol of the erythrocytes in the total lipid extraction were simultaneously measured by an HPLC method, and cholesterol was regarded as an internal standard for HPLC (29). Combination of the methods is relatively simple, and they are sensitive to the peroxidative changes of intact erythrocytes.…”

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confidence: 99%

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Effects of ascorbate on membrane phospholipids and tocopherols of intact erythrocytes during peroxidation by t‐butylhydroperoxide: Comparison with effects of dithiothreitol

Shiro

Murakami

2001

Lipids

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Peroxidation of intact human erythrocytes by t-butylhydroperoxide (tBHP)) was studied. By incubation of the erythrocytes with 1 mM tBHP, reduced glutathione (GSH) was exhausted within 1 min, and tocopherols (Toc) and phospholipids (PL) decreased to nearly their lowest levels (in this study) within 5 min. The rate of decrease of alpha-Toc was faster than that of gamma-Toc, but alpha-Toc was never exhausted. The rates of decrease of Toc were faster than that of PL. Malondialdehyde increased slowly to reach a maximal value at 30 min. Methemoglobin (metHB) reached a maximum at 15 min. The maximal levels of these substances were maintained until 90 min incubation, which indicated that the peroxidation by tBHP had stopped spontaneously until at least 90 min. By the incubation with tBHP for 30 min, phosphatidylethanolamine (PE) and alpha-Toc decreased to about 70 and 30% of control levels, respectively, and gamma-Toc and GSH were almost exhausted. Ascorbate (0.1 mM) afforded protection of 92% to PE, 50% to alpha-Toc, and 65% to gamma-Toc against peroxidation, but ascorbate had no preventive effect at all on the formation of metHB and the decrease of GSH. These results may indicate that ascorbate-mediated protection of the membrane PL against the peroxidation depends primarily on Toc. On the other hand, dithiothreitol (DTT) (5 mM) almost completely prevented the formation of metHB, and DTT completely protected the PL and Toc against peroxidation, indicating the importance of sulfhydryl groups in erythrocytes.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Effects of ascorbate on membrane phospholipids and tocopherols of intact erythrocytes during peroxidation by t‐butylhydroperoxide: Comparison with effects of dithiothreitol

Shiro

Murakami

2001

Lipids

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“…Cholesterol and Toc were measured simultaneously by the HPLC method as described in our previous reports. 13,14) The HPLC system (HP-1100, Yokogawa Electric Co., Tokyo, Japan) was connected in series to a ‰uorescence detector (FS-8020, Tosoh, Tokyo, Japan) and an evapolative light scattering detector (ELSD) (SEDEX-55, Vitry sur Seine, France).…”

Section: Methodsmentioning

confidence: 99%

High-cholesterol Diets Induce Changes in Lipid Composition of Rat Erythrocyte Membrane Including Decrease in Cholesterol, Increase in α-Tocopherol and Changes in Fatty Acids of Phospholipids

Shiro

Ohnishi

Kaji

et al. 2003

Bioscience, Biotechnology, and Biochemistry

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“…AA has been shown to play a protective role against peroxidation of RBC membrane lipids and tocopherols by tbutylhydroperoxide, preserving lipids by up to 92% and aand g-tocopherol by 50% and 65%, respectively (46). AA can also help protect a-TH from depletion in membranes treated with DABS (18), hence, protecting against lipid peroxidation (which occurs when a-TH levels are depleted by 20%).…”

Section: Protective Role Of Ascorbate For Rbc Membranesmentioning

confidence: 99%

Redox Reactions and Electron Transfer Across the Red Cell Membrane

Kennett

Kuchel

2003

IUBMB Life

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SummaryPlasma membrane electron transport systems appear to be ubiquitous. These systems are implicated in hormone signal transduction, cell growth and differentiation events as well as protection from oxidative stress. The red blood cell is constantly exposed to oxidative stress; protection against the reactive species generated during this process may be the main role of its membrane electron transport systems. Membrane redox activity has been studied for over three-quarters of a century, and yet many questions remain regarding its identity and likely roles: are electron transfers by distinct and specific mechanisms; what are the physiological donors and acceptors; and how do these systems affect metabolism? Current evidence suggests that the human erythrocyte membrane contains a number of distinct electron transfer systems, some of which, at least, involve membrane proteins, and NADH or ascorbate as electron donors. The activity of these systems appears to be closely related to the metabolic state of the cell, suggesting that mediation of reducing equivalents across the plasma membrane allows redox buffering of each environment, intra-and extracellular, by the other. We have decided to study this from a new perspective, NMR spectroscopy the area of our own technical expertise, hence this review is slanted towards this more recent analysis. IUBMB Life, 55: 375-385, 2003

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Effects of Ascorbic Acid on Peroxidation of Human Erythrocyte Membranes by Lipoxygenase.

Cited by 3 publications

References 35 publications

Effects of ascorbate on membrane phospholipids and tocopherols of intact erythrocytes during peroxidation by t‐butylhydroperoxide: Comparison with effects of dithiothreitol

Effects of ascorbate on membrane phospholipids and tocopherols of intact erythrocytes during peroxidation by t‐butylhydroperoxide: Comparison with effects of dithiothreitol

High-cholesterol Diets Induce Changes in Lipid Composition of Rat Erythrocyte Membrane Including Decrease in Cholesterol, Increase in α-Tocopherol and Changes in Fatty Acids of Phospholipids

Redox Reactions and Electron Transfer Across the Red Cell Membrane

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