Summary
To determine what biochemical indexes might be useful in measuring the endothelial response to hyperoxia in vitro we exposed endothelial cell monolayers (ECM) from pig aortas to either hyperoxic (95% O2:5% CO2, 1 atm) or control conditions (95% air: 5% CO2) and made of following measurements: (a) DNA and protein contents remaining in the ECM; (b) lactate dehydrogenase (LDH) activity in the medium; (c) the net uptake of rubidium (Rb+) adenine, and adenosine; and (d) cellular ATP and medium lactate.
Twelve hours of hyperoxic exposure did not cause significant changes. After 24 or 48 h of hyperoxia, DNA and protein contents were decreased; LDH activity and the protein-to-DNA ratio were increased; adenosine uptake was decreased per ECM but was unchanged when corrected for culture DNA and protein content. Adenine uptake was un-altered as were cellular ATP content and medium lactate concentration. The net Rb+ uptake-to-DNA ratio was increased after 24 h but not after 48 h of hyperoxia. The extent of the DNA and LDH changes indicated that the cellular disturbance caused by hyperoxia was progressive from 12 to 48 h.
Presence of superoxide dismutase (250 U/ml) prevented both the increase of LDH activity and the decrease of protein after 48 h but did not affect the decrease of DNA. These results suggest that the cells remaining in the ECM after hyperoxia have normal biochemical function and may represent a subpopulation of cells more resistant to oxygen toxicity than the damaged cells.
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This investigation was supported by Swiss National Science Foundation Grant 3.224.077 and by National Institutes of Health Grant HL00310-01.
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Rubin, D.B., Housset, B., Jean-Mairet, Y. et al. Effects of hyperoxia on biochemical indexes of pig aortic endothelial function. In Vitro 19, 625–634 (1983). https://doi.org/10.1007/BF02619576
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DOI: https://doi.org/10.1007/BF02619576