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Relation of fluorescence in lipid-containing red cell membrane extracts to in vivo lipid peroxidation
Abstract:
Previous studies have demonstrated that the abnormal fluorescent peak consistent with the crosslinking of red cell membrane constituents by the lipid-peroxide decomposition product MDA is demonstrable in lipid-containing extracts of red cells obtained from patients treated with the oxidizing hemolytic agent diaminodiphenylsulfone. The present studies were primarily aimed at ascertaining the specificity of this fluorescence as an indicator of in vivo red cell lipid peroxidation. Repetitive injection of phenylhydrazine or acetylphenylhydrazine in rats resulted in gradually increasing levels of fluorescence despite the return in hematocrit toward normal. Chromatography on Sephadex LH-20 of lipid-containing extracts obtained from the red cells of rats and rabbits treated with phenylhydrazine revealed fluorescent peaks similar to those observed in red cell lipid incubated with MDA. In addition, increased levels of fluorescence were observed in the extracts of red cells obtained from three vitamin E--deficient premature infants. These findings suggest that the fluorescence assay is a sensitive indicator of in vivo red cell lipid peroxidation.
Insights
This study shows that a specific fluorescence assay can detect lipid peroxidation in red blood cells. This method is sensitive and indicates oxidative damage in vivo.
Area of Science:
- Biochemistry
- Hematology
- Toxicology
Background:
- Abnormal fluorescence in red blood cells is linked to lipid peroxidation.
- Diaminodiphenylsulfone treatment causes red cell membrane crosslinking via MDA.
Purpose of the Study:
- To determine if red cell fluorescence indicates in vivo lipid peroxidation.
- To validate a fluorescence assay for detecting oxidative damage.
Main Methods:
- Inducing lipid peroxidation in rats and rabbits using phenylhydrazine/acetylphenylhydrazine.
- Analyzing red cell lipid extracts using Sephadex LH-20 chromatography.
- Measuring fluorescence in red cells from vitamin E-deficient infants.
Main Results:
- Repetitive injections increased red cell fluorescence in rats, even as hematocrit normalized.
- Chromatography revealed MDA-like fluorescent peaks in treated animals.
- Elevated fluorescence was found in vitamin E-deficient premature infants' red cells.
Conclusions:
- The fluorescence assay is a sensitive indicator of in vivo red cell lipid peroxidation.
- This assay can detect oxidative damage in various conditions.