Concomitant administration of hydralazine completely prevented the development of nitroglycerin tolerance in rabbits and normalized vascular superoxide production by inhibiting a membrane-bound NADH oxidase.
Does hydralazine prevent nitrate tolerance and reduce vascular superoxide production in nitroglycerin-treated rabbits?
Hydralazine prevents nitroglycerin tolerance by inhibiting the activation of a membrane-bound NADH oxidase, providing a mechanistic basis for its clinical benefits when used with nitrates.
Absolute Event Rate: 477% vs 3123%
p-value: p=<0.05
Hydralazine has been shown to reduce mortality in patients with congestive heart failure when given concomitantly with isosorbide dinitrate. Recently, we demonstrated that nitrate tolerance is in part due to enhanced vascular superoxide .O2- production. We sought to determine mechanisms whereby hydralazine may prevent tolerance. Rabbits either received no treatment, nitroglycerin patches (1.5 micrograms/kg/min x 3 d), hydralazine alone (10 mg/kg/d in drinking water), or hydralazine and nitroglycerin. Aortic segments were studied in organ chambers and relative rates of vascular .O2- production were determined using lucigenin-enhanced chemiluminescence. Nitroglycerin treatment markedly inhibited relaxations to nitroglycerin (maximum relaxations in untreated: 92 +/- 1 vs. 64 +/- 3% in nitroglycerin-treated patients and increased vascular .O2- production by over two-fold (P < 0.05). Treatment with hydralazine in rabbits not receiving nitroglycerin significantly decreased .O2- production in intact rabbit aorta and increased sensitivity to nitroglycerin. When given concomitantly with nitroglycerin, hydralazine completely prevented the development of nitrate tolerance and normalized endogenous rates of vascular .O2- production. Studies of vessel homogenates demonstrated that the major source of .O2- was an NADH-dependent membrane-associated oxidase displaying activities of 67 +/- 12 vs. 28 +/- 2 nmol .O2-.min-1.mg protein-1 in nitroglycerin-treated vs. untreated aortic homogenates. In additional studies, we found that acute addition of hydralazine (10 microM) to nitroglycerin-tolerant vessels immediately inhibited .O2- production and NADH oxidase activity in vascular homogenates. The chemiluminescence signal was inhibited by a recombinant heparin-binding superoxide dismutase (HBSOD) demonstrating the specificity of this assay for .O2-. These observations suggest that a specific membrane-associated oxidase is activated by chronic nitroglycerin treatment, and the activity of this oxidase is inhibited by hydralazine, providing a mechanism whereby hydralazine may prevent tolerance. The ability of hydralazine to inhibit vascular .O2- anion production represents a novel mechanism of action for this drug.
Münzel et al. (1996) studied Nitroglycerin tolerance. Hydralazine vs. Nitroglycerin alone (1.5 μg/kg/min patch) or no treatment was evaluated on Vascular superoxide production (lucigenin counts/mg/min) (p=<0.05). Concomitant administration of hydralazine completely prevented the development of nitroglycerin tolerance in rabbits and normalized vascular superoxide production by inhibiting a membrane-bound NADH oxidase.