Enhanced ADP-ribosylation and its diminution by lipoamide after ischemia-reperfusion in perfused rat heart

Eszter Szabados, Gabor M. Fischer, Ferenc Gallyas, Gyula Kispal, Balazs Sumegi

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Abstract

Poly-ADP-ribose polymerase (PARP) is considered to play an important role in oxidative cell damage. We assumed that ischemia-reperfusion resulting from the increasing reactive oxygen species (ROS) can lead to the activation of endogenous mono- and poly-ADP-ribosylation reactions and that the reduction of ROS level by lipoamide, a less known antioxidant, can reverse these unfavorable processes. Experiments were performed on isolated Langendorff hearts subjected to 60-min ischemia followed by reperfusion. ROS, malondialdehyde, deoxyribonucleic acid (DNA) breaks, and NAD+ content were assayed in the hearts, and the ADP-ribosylation of cytoplasmic and nuclear proteins were determined by Western blot assay. Ischemia-reperfusion caused a moderate (30.2 ± 8%) increase in ROS production determined by the dihydrorhodamine123 method and significantly increased the malondialdehyde production (from <1 to 23 ± 2.7 nmol/ml), DNA damage (undamaged DNA decreased from 71 ± 7% to 23.1 ± 5%), and NAD+ catabolism. In addition, ischemia-reperfusion activated the mono-ADP-ribosylation of GRP78 and the self-ADP-ribosylation of the nuclear PARP. The perfusion of hearts with lipoamide significantly decreased the ischemia-reperfusion-induced cell membrane damage determined by enzyme release (LDH, CK, and GOT), decreased the ROS production, reduced the malondialdehyde production to 5.5 ± 2.4 nmol/ml, abolished DNA damage, and reduced NAD+ catabolism. The ischemia-reperfusion-induced activation of poly- and mono-ADP-ribosylation reactions were also reverted by lipoamide. In isolated rat heart mitochondria, dihydrolipoamide was found to be a better antioxidant than dihydrolipoic acid. Ischemia-reperfusion by ROS overproduction and increasing DNA breaks activates PARP leading to accelerated NAD+ catabolism, impaired energy metabolism, and cell damage. Lipoamide by reducing ROS levels halts PARP activation and membrane damage and improves the recovery of postischemic myocardium. Copyright (C) 1999 Elsevier Science Inc.

Original languageEnglish
Pages (from-to)1103-1113
Number of pages11
JournalFree Radical Biology and Medicine
Volume27
Issue number9-10
DOIs
Publication statusPublished - Nov 1 1999

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Keywords

  • ADP-ribosylation
  • Antioxidant
  • Cell damage
  • Chaperone
  • Folding
  • Free radicals
  • GRP78
  • Heart perfusion
  • Ischemia-reperfusion
  • Lipid peroxidation
  • Lipoamide
  • Poly-ADP-ribose polymerase
  • Protein transport
  • Reactive oxygen species
  • Signaling

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

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