Glucose-6-phosphate dehydrogenase-derived NADPH fuels superoxide production in the failing heart.
In the failing heart, NADPH oxidase and uncoupled NO synthase utilize cytosolic NADPH to form superoxide. NADPH is supplied principally by the pentose phosphate pathway, whose rate-limiting enzyme is glucose 6-phosphate dehydrogenase (G6PD). Therefore, we hypothesized that cardiac G6PD activation drives part of the excessive superoxide production implicated in the pathogenesis of heart failure. Pacing-induced heart failure was performed in eight chronically instrumented dogs. Seven normal dogs served as control. End-stage failure occurred after 28 +/- 1 days of pacing, when left ventricular end-diastolic pressure reached 25 mm Hg. In left ventricular tissue homogenates, spontaneous superoxide generation measured by lucigenin (5 microM) chemiluminescence was markedly increased in heart failure (1338 +/- 419 vs. 419 +/- 102 AU/mg protein, P < 0.05), as were NADPH levels (15.4 +/- 1.5 vs. 7.5 +/- 1.5 micromol/gww, P < 0.05). Superoxide production was further stimulated by the addition of NADPH. The NADPH oxidase inhibitor gp91(ds-tat) (50 microM) and the NO synthase inhibitor L-NAME (1 mM) both significantly lowered superoxide generation in failing heart homogenates by 80% and 76%, respectively. G6PD was upregulated and its activity higher in heart failure compared to control (0.61 +/- 0.10 vs. 0.24 +/- 0.03 nmol/min/mg protein, P < 0.05), while superoxide production decreased to normal levels in the presence of the G6PD inhibitor 6-aminonicotinamide. We conclude that the activation of myocardial G6PD is a novel mechanism that enhances NADPH availability and fuels superoxide-generating enzymes in heart failure.
Gupte
Sachin A
SA
Department of Physiology, BSB Room 626, New York Medical College, Valhalla, 10595, USA. sachin_gupte@nymc.edu
Levine
Robert J
RJ
Gupte
Rakhee S
RS
Young
Martin E
ME
Lionetti
Vincenzo
V
Labinskyy
Volodymyr
V
Floyd
Beverly C
BC
Ojaimi
Caroline
C
Bellomo
Michelle
M
Wolin
Michael S
MS
Recchia
Fabio A
FA
D015120:6-Aminonicotinamide / Q000494:pharmacology; D000818:Animals; D001794:Blood Pressure; D002304:Cardiac Pacing, Artificial / Q000009:adverse effects; D004195:Disease Models, Animal; D004285:Dogs; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006333:Heart Failure / Q000201:enzymology* / Q000209:etiology / Q000503:physiopathology; D006801:Humans; D009249:NADP / Q000096:biosynthesis*; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*; D013723:Teratogens / Q000494:pharmacology; D013997:Time Factors
D004791:Enzyme Inhibitors; D013723:Teratogens; D013481:Superoxides; D015120:6-Aminonicotinamide; D009249:NADP; D005954:Glucosephosphate Dehydrogenase; D019001:Nitric Oxide Synthase; D019331:NG-Nitroarginine Methyl Ester