TY - JOUR
T1 - Poly(ADP-ribose) polymerase inhibition improves endothelial dysfunction induced by reactive oxidant hydrogen peroxide in vitro
AU - Radovits, Tamás
AU - Lin, Li ni
AU - Zotkina, Julia
AU - Gero, Domokos
AU - Szabó, Csaba
AU - Karck, Matthias
AU - Szabó, Gábor
N1 - Funding Information:
This work was supported by a grant from the German Research Foundation (SFB 414) to G.S. and by the Hungarian Research Fund (OTKA AT049488) and the National Institutes of Health (R01 GM060915) to C.S. The expert technical assistance of Anne Schuppe and Heike Ziebart are gratefully acknowledged.
PY - 2007/6/14
Y1 - 2007/6/14
N2 - Reactive oxygen species, such as hydrogen peroxide (H2O2) induce oxidative stress and DNA-injury. The subsequent activation of poly(ADP-ribose) polymerase (PARP) has been implicated in the pathogenesis of various cardiovascular diseases including ischaemia-reperfusion injury, circulatory shock, diabetic complications and atherosclerosis. We investigated the effect of PARP-inhibition on endothelial dysfunction induced by H2O2. In vascular reactivity measurements on isolated rat aortic rings we investigated the phenylephrine-induced contraction, and endothelium-dependent and -independent vasorelaxation by using cumulative concentrations of acetylcholine and sodium nitroprusside. Endothelial dysfunction was induced by exposing the rings to H2O2 (200 and 400 μM) for 30 min. In the treatment group, rings were preincubated with the potent PARP-inhibitor INO-1001. DNA strand breaks were assessed by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method. Immunohistochemical analysis was performed for poly(ADP-ribose) (the enzymatic product of PARP) and for apoptosis inducing factor (a pro-apoptotic factor regulated by PARP). Exposure to H2O2 resulted in reduced contraction forces and a dose-dependent impairment of endothelium-dependent vasorelaxation of aortic rings (maximal relaxation to acetylcholine: 86.21 ± 1.574% control vs. 72.55 ± 1.984% H2O2 200 μM vs. 66.86 ± 1.961% H2O2 400 μM; P < 0.05). PARP-inhibition significantly improved the acetylcholine-induced vasorelaxation (77.75 ± 3.019% vs. 66.86 ± 1.961%; P < 0.05), while the contractility remained unaffected. The dose-response curves of endothelium-independent vasorelaxation to sodium nitroprusside did not differ in any groups studied. In the H2O2 groups immunohistochemical analysis showed enhanced PARP-activation and nuclear translocation of apoptosis inducing factor, which were prevented by INO-1001. Our results demonstrate that PARP activation contributes to the pathogenesis of H2O2-induced endothelial dysfunction, which can be prevented by PARP inhibitors.
AB - Reactive oxygen species, such as hydrogen peroxide (H2O2) induce oxidative stress and DNA-injury. The subsequent activation of poly(ADP-ribose) polymerase (PARP) has been implicated in the pathogenesis of various cardiovascular diseases including ischaemia-reperfusion injury, circulatory shock, diabetic complications and atherosclerosis. We investigated the effect of PARP-inhibition on endothelial dysfunction induced by H2O2. In vascular reactivity measurements on isolated rat aortic rings we investigated the phenylephrine-induced contraction, and endothelium-dependent and -independent vasorelaxation by using cumulative concentrations of acetylcholine and sodium nitroprusside. Endothelial dysfunction was induced by exposing the rings to H2O2 (200 and 400 μM) for 30 min. In the treatment group, rings were preincubated with the potent PARP-inhibitor INO-1001. DNA strand breaks were assessed by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method. Immunohistochemical analysis was performed for poly(ADP-ribose) (the enzymatic product of PARP) and for apoptosis inducing factor (a pro-apoptotic factor regulated by PARP). Exposure to H2O2 resulted in reduced contraction forces and a dose-dependent impairment of endothelium-dependent vasorelaxation of aortic rings (maximal relaxation to acetylcholine: 86.21 ± 1.574% control vs. 72.55 ± 1.984% H2O2 200 μM vs. 66.86 ± 1.961% H2O2 400 μM; P < 0.05). PARP-inhibition significantly improved the acetylcholine-induced vasorelaxation (77.75 ± 3.019% vs. 66.86 ± 1.961%; P < 0.05), while the contractility remained unaffected. The dose-response curves of endothelium-independent vasorelaxation to sodium nitroprusside did not differ in any groups studied. In the H2O2 groups immunohistochemical analysis showed enhanced PARP-activation and nuclear translocation of apoptosis inducing factor, which were prevented by INO-1001. Our results demonstrate that PARP activation contributes to the pathogenesis of H2O2-induced endothelial dysfunction, which can be prevented by PARP inhibitors.
KW - Apoptosis
KW - Endothelial dysfunction
KW - Hydrogen peroxide
KW - Oxidative stress
KW - Poly(ADP-ribose) polymerase
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U2 - 10.1016/j.ejphar.2007.02.060
DO - 10.1016/j.ejphar.2007.02.060
M3 - Article
C2 - 17397824
AN - SCOPUS:34250648890
SN - 0014-2999
VL - 564
SP - 158
EP - 166
JO - European Journal of Pharmacology
JF - European Journal of Pharmacology
IS - 1-3
ER -