Role of lipid peroxidation and poly(ADP-ribose) polymerase activation in oxidant-induced membrane transport dysfunction in opossum kidney cells.
This study was undertaken to examine the role of lipid peroxidation and poly(ADP-ribose) polymerase (PARP) activation in H(2)O(2)-induced inhibition of Na(+)-dependent phosphate (Na(+)-Pi) uptake in opossum kidney (OK) cells. H(2)O(2) inhibited Na(+)-Pi uptake in a dose-dependent manner. H(2)O(2)-induced inhibition of Na(+)-Pi uptake was prevented by dithiothreitol and glutathione. A potent antioxidant, DPPD, had no effect on H(2)O(2) inhibition of Na(+)-Pi uptake, despite completely inhibiting lipid peroxidation induced by H(2)O(2). However, in primary cultured rabbit proximal tubular cells, the effect of H(2)O(2) on Na(+)-Pi uptake was significantly prevented by DPPD, suggesting a species difference in the role of lipid peroxidation in the inhibition of Na(+)-Pi uptake occurring with H(2)O(2). t-Butylhydroperoxide (tBHP) caused the inhibition of Na(+)-Pi uptake that was prevented by DPPD in OK cells and rabbit proximal tubular cells. The PARP inhibitor 3-aminobenzamide completely protected the inhibition of Na(+)-Pi uptake induced by H(2)O(2) but not by tBHP. H(2)O(2)-induced ATP depletion was prevented by 3-aminobenzamide but not by DPPD. tBHP-induced ATP depletion was prevented by DPPD, whereas it was not altered by 3-aminobenzamide. Effects of H(2)O(2) and tBHP on Na(+)-Pi uptake and ATP depletion were prevented by an iron chelator, deferoxamine, suggesting that the oxidants inhibit Na(+)-Pi uptake through an iron-dependent mechanism. The extent of DNA damage by tBHP was similar to that by H(2)O(2). These results indicate that the effect of H(2)O(2) on membrane transport function in OK cells is associated with PARP activation but not lipid peroxidation, whereas the effect of tBHP is associated with lipid peroxidation.
Min
S K
SK
Department of Pediatrics, Pusan National University, Pusan, 602-739, Korea.
Kim
S Y
SY
Kim
C H
CH
Woo
J S
JS
Jung
J S
JS
Kim
Y K
YK
D000818:Animals; D001549:Benzamides / Q000494:pharmacology; D001692:Biological Transport / Q000187:drug effects; D002478:Cells, Cultured; D004229:Dithiothreitol / Q000494:pharmacology; D004789:Enzyme Activation; D005978:Glutathione / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000633:toxicity*; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism; D015227:Lipid Peroxidation; D009893:Opossums; D010710:Phosphates / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000502:physiology*; D011817:Rabbits; D012964:Sodium / Q000378:metabolism
D001549:Benzamides; D010710:Phosphates; C025160:3-aminobenzamide; D012964:Sodium; D006861:Hydrogen Peroxide; D011065:Poly(ADP-ribose) Polymerases; D005978:Glutathione; D004229:Dithiothreitol