Mechanisms of endothelial cell ATP depletion after oxidant injury

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Abstract

To investigate mechanisms of ATP depletion in human umbilical vein endothelial cells after oxidant injury, we studied the relationship between DNA damage, activation of the DNA-repairing enzyme poly ADP-ribose polymerase, NAD depletion, and ATP depletion. We found that oxidant stress generated with hypoxanthine-xanthine oxidase and glucose-glucose oxidase resulted in profound DNA damage. When endothelial cells were exposed to 25 and 50 mU/ml xanthine oxidase for 60 min, the percentage of double-stranded DNA was significantly reduced (p < 0.05) to 15.2 ± 1.2 and 4.6 ± 0.5%, respectively, compared to 75.7 ± 3.9% for control cells. When endothelial cells were exposed to 25 and 50 mU/ml glucose oxidase for 60 min, the percentage of double-stranded DNA was significantly (p < 0.05) reduced to 35.0 ± 1.5% and 9.9 ± 7.7%, respectively, compared to 73.2 ± 2.4% for control cells. ATP and NAD levels declined simultaneously with DNA damage. Because activation of the DNA-repairing enzyme poly ADP-ribose polymerase can consume NAD sufficient to interfere with ATP synthesis, we studied NAD and ATP levels after oxidant injury when ADP-ribose polymerase was inhibited with 3-aminobenzamide and nicotinamide. When poly ADP-ribose polymerase was inhibited, NAD levels remained normal, but ATP depletion was not prevented. We conclude that oxidant injury to human umbilical vein endothelial cells results in profound DNA damage and NAD and ATP depletion. NAD depletion results from activation of poly ADP-ribose polymerase, but this phenomenon is not the mechanism of ATP depletion in human umbilical vein endothelial cells.

Original languageEnglish
Pages (from-to)97-101
Number of pages5
JournalPediatric Research
Volume25
Issue number1
StatePublished - 1989

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Oxidants
NAD
Endothelial Cells
Adenosine Triphosphate
Poly(ADP-ribose) Polymerases
Wounds and Injuries
DNA Damage
Human Umbilical Vein Endothelial Cells
Glucose Oxidase
Xanthine Oxidase
DNA
Adenosine Diphosphate Ribose
Niacinamide
Enzymes
Glucose

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health

Cite this

Mechanisms of endothelial cell ATP depletion after oxidant injury. / Andreoli, Sharon.

In: Pediatric Research, Vol. 25, No. 1, 1989, p. 97-101.

Research output: Contribution to journalArticle

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