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Detection of a ferrylhemoglobin intermediate in an endothelial cell model after hypoxia-reoxygenation
Division of Hematology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892 A cell culture model of bovine aortic endothelial cells attached to microcarrier beads was used to study the interaction of diaspirin cross-linked hemoglobin (an oxygen-carryi...
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| Published in: | American journal of physiology. Heart and circulatory physiology 1999-07, Vol.277 (1), p.H92-H99 |
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| Main Authors: | , |
| Format: | Article |
| Language: | English |
| Citations: | Items that this one cites Items that cite this one |
| Online Access: | Get full text |
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| Summary: | Division of Hematology, Center for Biologics Evaluation and
Research, Food and Drug Administration, Bethesda, Maryland 20892
A cell culture model
of bovine aortic endothelial cells attached to microcarrier beads was
used to study the interaction of diaspirin cross-linked hemoglobin (an
oxygen-carrying blood substitute) with hypoxia-reoxygenation.
Hemoglobin (200 µM) and hypoxia-volume restriction (3-5 h),
together and separately, caused toxicity in this model, as measured by
decreased cellular replating efficiency. Hemoglobin (60 µM) caused a
reduction in hydrogen peroxide concentration and an increase in lipid
peroxidation above that induced by hypoxia alone. Incubation of
hemoglobin with endothelial cells caused transient oxidation of
hemoglobin to its highly reactive and toxic ferryl species after 3 h
of hypoxia, followed by 1 h of reoxygenation. Lipid peroxidation, which
may occur in the presence of ferrylhemoglobin, also occurred after 1 h
of reoxygenation. Hemoglobin caused a dose-dependent decrease in
intracellular glutathione concentration, suggesting that it caused an
oxidative stress to the cells. However, addition of ascorbate,
-tocopherol, or trolox did not decrease hemoglobin oxidation in the
presence of normal or hypoxic cells. It is concluded that diaspirin
cross-linked hemoglobin forms a ferryl intermediate in the absence of
any exogenously added oxidant and contributes to the oxidative burden
experienced by endothelial cells after hypoxia-reoxygenation, a
condition that is likely to be encountered during trauma and surgery
when hemoglobin solutions are used as perfusion agents.
hemoglobin-based oxygen carriers; endothelium; antioxidants |
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| ISSN: | 0363-6135 1522-1539 |
| DOI: | 10.1152/ajpheart.1999.277.1.h92 |