Abstract
Oxidants, induce injury and death of neighboring cells. We have studied early changes following exposure of the cells to oxidants. Exposure of P388D1 cells to H2O2 (0.1 to 2.5 mM) induced a rapid (3-5 min) fall in ATP, turnover of components of the glutathione cycle, a slower (>15 min) rise in intracellular Ca++ and Na+ and, after 4-5 hrs, cell lysis. Exposure to H2O2 produced a slight elevation of ADP at a time when ATP levels were low, and a stoichiometric increase in IMP and inosine. In determining the cause for the fall in ATP, we observed that P388D1 cells exposed to 0.04 to 2.5 mM H2O2 lost NAD at an initial rate of 0.26 nmoles/106 cells/min, leading to an 80 percent depletion of NAD within 20 min. This fall in NAD is coupled with inhibition of glycolysis. Poly(ADP-ribose)polymerase, the nuclear enzyme catalyzing conversion of NAD to nicotinamide and protein-bound ADP-ribose, was activated by exposure of the cells to H2O2. Inhibition of this enzyme by 2.5 mM 3-amionobenzamide prevented the oxidant induced fall in NAD. This also prevented the sustained fall in ATP, and the other events (above) except for glutathione cycle activity. H2O2 induced DNA strand breaks in target cells, potentially responsible for activation of the polymerase. These cells indicate that oxidant induced stimulation of poly(ADP-ribose)polymerase causes a fall in NAD leading to a sustained depletion of ATP, leading to cell death.
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Cochrane, C., Schraufstätter, I., Hinshaw, D. et al. ATP IN OXIDANT-INDUCED TISSUE INJURY: 35. Pediatr Res 19, 749 (1985). https://doi.org/10.1203/00006450-198507000-00055
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DOI: https://doi.org/10.1203/00006450-198507000-00055