Nitroblue tetrazolium inhibits oxidation of glyceryl trinitrate to nitric oxide in bovine aortic smooth muscle cells

Alessandra Pistelli, Vincenzo Mollace, Gluseppe Nistico, Daniela Salvemini, John Vane

Research output: Contribution to journalArticlepeer-review

Abstract

The effects of nitroblue tetrazolium (NBT), a well-known scavenger of Superoxide anions and an inhibitor of nicotinamide adenine dinucleotide (NADPH)-dependent oxidations, were assessed on the metabolism of glyceryl trinitrate (GTN) to nitric oxide (NO) by bovine aortic smooth muscle cells (SMC). The extent of this metabolism was determined by measuring NO formed, using the inhibition of thrombin-induced platelet aggregation and relaxation of rabbit aortic strips as bioassay systems. In addition, NO produced from GTN by SMC was measured as nitrite (NO2-), one of its breakdown products. The antiplatelet effect of GTN (44 μM) was potentiated by SMC (0.12-0.46 × 105 cells) treated with indomethacin (10 μM) and this was inhibited in a concentration-dependent manner when the cells were pretreated with NBT (100 μM). NBT (3-100 μM) also reduced the formation of NO2- from GTN (600 μM) by SMC (3 × 105 cells). Furthermore, relaxations of endothelium-denuded strips of the rabbit aorta by GTN (10-9-10-6 M) were attenuated when the strips were pretreated with NBT (100 or 500 μM). The formation of NO from l-arginine (l-Arg) by SMC was not affected by NBT. The hypotensive responses to GTN (0.25-1 mg/kg, i.v.) in anaesthetized rats were inhibited by pretreatment with NBT (1.25 mg/kg, i.v.) but NBT did not alter the hypotensive responses induced by SIN-1. Thus, NBT inhibited the bioconversion of GTN to NO both in vitro and in vivo. NBT may be a useful pharmacological tool to investigate the enzymic pathway(s) involved in the conversion of GTN to NO by smooth muscle cells or other cells.

Original languageEnglish
Pages (from-to)1737-1742
Number of pages6
JournalBiochemical Pharmacology
Volume47
Issue number10
DOIs
Publication statusPublished - May 18 1994

Keywords

  • glyceryl trinitrate
  • nitric oxide
  • nitroblue tetrazolium
  • platelets
  • smooth muscle

ASJC Scopus subject areas

  • Pharmacology

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