TY - JOUR
T1 - The positive inotropic effect of endothelin-1 is mediated by mitochondrial reactive oxygen species
AU - De Giusti, V. C.
AU - Correa, M. V.
AU - Villa-Abrille, M. C.
AU - Beltrano, C.
AU - Yeves, A. M.
AU - de Cingolani, G. E.Chiappe
AU - Cingolani, H. E.
AU - Aiello, E. A.
N1 - Funding Information:
This study was partly supported by grants (PICT 05-08512 to HEC, PICT 05-12480 to GECC and PICT 25495 to EAA) of the Agencia Nacional de Promoción Científica y Tecnológica de Argentina. HOE642 was kindly provided by Aventis Pharma (Germany).
Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.
PY - 2008/8/15
Y1 - 2008/8/15
N2 - We have previously demonstrated the participation of reactive oxygen species (ROS) in the positive inotropic effect of a physiological concentration of Angiotensin II (Ang II, 1 nM). The objective of the present work was to evaluate the role and source of ROS generation in the positive inotropic effect produced by an equipotent concentration of endothelin-1 (ET-1, 0.4 nM). Isolated cat ventricular myocytes were used to measure sarcomere shortening with a video-camera, superoxide anion ({radical dot}O2-) with chemiluminescence, and ROS production and intracellular pH (pHi) with epifluorescence. The ET-1-induced positive inotropic effect (40.4 ± 3.1%, n = 10, p < 0.05) was associated to an increase in ROS production (105 ± 29 fluorescence units above control, n = 6, p < 0.05). ET-1 also induced an increase in {radical dot}O2- production that was inhibited by the NADPH oxidase blocker, apocynin, and by the blockers of mitochondrial ATP-sensitive K+ channels (mKATP), glibenclamide and 5 hydroxydecanoic acid. The ET-1-induced positive inotropic effect was inhibited by apocynin (0.3 mM; 6.3 ± 6.6%, n = 13), glibenclamide (50 μM; 8.8 ± 3.5%, n = 6), 5 hydroxydecanoic acid (500 μM; 14.1 ± 8.1, n = 9), and by scavenging ROS with MPG (2 mM; 0.92 ± 5.6%, n = 8). ET-1 enhanced proton efflux (JH) carried by the Na+/H+ exchanger (NHE) after an acid load, effect that was blocked by MPG. Consistently, the ET-induced positive inotropic effect was also inhibited by the NHE selective blocker HOE642 (5 μM; 9.37 ± 6.07%, n = 7). The data show that the effect of a concentration of ET-1 that induces an increase in contractility of about 40% is totally mediated by an intracellular pathway triggered by mitochondrial ROS formation and stimulation of the NHE.
AB - We have previously demonstrated the participation of reactive oxygen species (ROS) in the positive inotropic effect of a physiological concentration of Angiotensin II (Ang II, 1 nM). The objective of the present work was to evaluate the role and source of ROS generation in the positive inotropic effect produced by an equipotent concentration of endothelin-1 (ET-1, 0.4 nM). Isolated cat ventricular myocytes were used to measure sarcomere shortening with a video-camera, superoxide anion ({radical dot}O2-) with chemiluminescence, and ROS production and intracellular pH (pHi) with epifluorescence. The ET-1-induced positive inotropic effect (40.4 ± 3.1%, n = 10, p < 0.05) was associated to an increase in ROS production (105 ± 29 fluorescence units above control, n = 6, p < 0.05). ET-1 also induced an increase in {radical dot}O2- production that was inhibited by the NADPH oxidase blocker, apocynin, and by the blockers of mitochondrial ATP-sensitive K+ channels (mKATP), glibenclamide and 5 hydroxydecanoic acid. The ET-1-induced positive inotropic effect was inhibited by apocynin (0.3 mM; 6.3 ± 6.6%, n = 13), glibenclamide (50 μM; 8.8 ± 3.5%, n = 6), 5 hydroxydecanoic acid (500 μM; 14.1 ± 8.1, n = 9), and by scavenging ROS with MPG (2 mM; 0.92 ± 5.6%, n = 8). ET-1 enhanced proton efflux (JH) carried by the Na+/H+ exchanger (NHE) after an acid load, effect that was blocked by MPG. Consistently, the ET-induced positive inotropic effect was also inhibited by the NHE selective blocker HOE642 (5 μM; 9.37 ± 6.07%, n = 7). The data show that the effect of a concentration of ET-1 that induces an increase in contractility of about 40% is totally mediated by an intracellular pathway triggered by mitochondrial ROS formation and stimulation of the NHE.
KW - Cardiac myocytes
KW - Endothelin-1
KW - Inotropic effect
KW - Reactive oxygen species
KW - Sodium-hydrogen exchanger
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U2 - 10.1016/j.lfs.2008.06.008
DO - 10.1016/j.lfs.2008.06.008
M3 - Article
C2 - 18625248
AN - SCOPUS:48549106437
SN - 0024-3205
VL - 83
SP - 264
EP - 271
JO - Life Sciences
JF - Life Sciences
IS - 7-8
ER -