Inhibitors of poly (ADP-ribose) synthetase protect rat proximal tubular cells against oxidant stress.

Abstract

BACKGROUND:

The generation of reactive oxygen species (ROS) has been implicated in the pathogenesis of renal ischemia-reperfusion injury. ROS produce DNA strand breaks that lead to the activation of the DNA-repair enzyme poly (ADP-ribose) synthetase (PARS). Excessive PARS activation results in the depletion of its substrate, nicotinamide adenine dinucleotide (NAD) and subsequently of adenosine 5'-triphosphate (ATP), leading to cellular dysfunction and eventual cell death. The aim of this study was to investigate the effect of various PARS inhibitors on the cellular injury and death of rat renal proximal tubular (PT) cells exposed to hydrogen peroxide (H2O2).

METHODS:

Rat PT cell cultures were incubated with H2O2 (1 mM) either in the presence or absence of the PARS inhibitors 3-aminobenzamide (3-AB, 3 mM), 1,5-dihydroxyisoquinoline (0.3 mM) or nicotinamide (Nic, 3 mM), or increasing concentrations of desferrioxamine (0.03 to 3 mM) or catalase (0.03 to 3 U/ml). Cellular injury and death were determined using the MTT and lactate dehydrogenase (LDH) assays, respectively. H2O2-mediated PARS activation in rat PT cells and the effects of PARS inhibitors on PARS activity were determined by measurement of the incorporation of [3H]NAD into nuclear proteins.

RESULTS:

Incubation of rat PT cells with H2O2 significantly inhibited mitochondrial respiration and increased LDH release, respectively. Both desferrioxamine and catalase reduced H2O2-mediated cellular injury and death. All three PARS inhibitors significantly attenuated the H2O2-mediated decrease in mitochondrial respiration and the increase in LDH release. Incubation with H2O2 produced a significant increase in PARS activity that was significantly reduced by all PARS inhibitors. 3-Aminobenzoic acid (3 mM) and nicotinic acid (3 mM), structural analogs of 3-AB and Nic, respectively, which did not inhibit PARS activity, did not reduce the H2O2-mediated injury and necrosis in cultures of rat PT cells.

CONCLUSION:

We propose that PARS activation contributes to ROS-mediated injury of rat PT cells and, therefore, to the cellular injury and cell death associated with conditions of oxidant stress in the kidney.

• Authors: P K Chatterjee, S Cuzzocrea, C Thiemermann
• Journal: Kidney international (Kidney Int) Vol. 56 Issue 3 Pg. 973-84 (Sep 1999) ISSN: 0085-2538 [Print] United States
• PMID: 10469365 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Benzamides
• Enzyme Inhibitors
• Isoquinolines
• Poly(ADP-ribose) Polymerase Inhibitors
• Niacinamide
• 1,5-dihydroxyisoquinoline
• 3-aminobenzamide
• Hydrogen Peroxide
• L-Lactate Dehydrogenase
• Catalase
• Deferoxamine

Topics

• Animals
• Benzamides (pharmacology)
• Catalase (pharmacology)
• Cells, Cultured
• DNA Damage
• Deferoxamine (pharmacology)
• Enzyme Activation (drug effects)
• Enzyme Inhibitors (pharmacology)
• Hydrogen Peroxide (toxicity)
• Isoquinolines (pharmacology)
• Kidney Tubules, Proximal (cytology, drug effects, metabolism)
• L-Lactate Dehydrogenase (metabolism)
• Necrosis
• Niacinamide (pharmacology)
• Oxidative Stress (drug effects)
• Poly(ADP-ribose) Polymerase Inhibitors
• Rats