Effect of isosorbide dinitrate on nitric oxide synthase under hypoxia.
Nitric oxide synthase (NOS) catalyzes nitric oxide (NO) formation from L-arginine in the presence of molecular oxygen and NADPH. NO is involved in the regulation of microvasculature. Isosorbide dinitrate (ISDN) and glyceryl trinitrate (GTN) have been widely used as vasodilators to treat acute myocardial ischemia, their biological effects being due to the release of NO. In this investigation, the effects of ISDN and GTN on NOS activity in the presence or absence of oxyhemoglobin under hypoxia and normoxia were studied. The apparent K(m) values for molecular oxygen were 21.6 +/- 1.5 and 9.4 +/- 1.3 micromol/l for nNOS and eNOS, respectively. ISDN liberated NO in a concentration- and pH-dependent manner, but no differences between hypoxia and normoxia were observed. The NO release from ISDN was also measured directly by an electron spin resonance spectral method with N-(dithiocarboxy)sarcosine-Fe complex as a NO-trapping agent. ISDN increased nNOS and eNOS activities in the presence of 30 micromol/l oxyhemoglobin under hypoxia, while it did not affect nNOS and eNOS activities under normoxia. In the absence of oxyhemoglobin, ISDN inhibited nNOS and eNOS activities under both hypoxic and normoxic experimental conditions. The rate of oxygen release from oxyhemoglobin under hypoxia was increased 3 times in the presence of 1 mmol/l ISDN. In contrast to ISDN, GTN could not release NO spontaneously, and it also did not affect nNOS and eNOS activities in the absence or presence of 30 micromol/l oxyhemoglobin under both hypoxic and normoxic conditions. These results indicated that the NO release from ISDN is different from that of GTN, and the increase of NOS activity by ISDN in the presence of oxyhemoglobin under hypoxia is ascribed to the increase in molecular oxygen concentration.
Jiang
H B
HB
Department of Biochemistry, Kagawa Medical University, Kagawa, Japan. hjiang@kms.ac.jp
Yoneyama
H
H
Furukawa
A
A
Hamamoto
T
T
Takahara
J
J
Ichikawa
Y
Y
D000818:Animals; D002956:Citrulline / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006863:Hydrogen-Ion Concentration; D007548:Isosorbide Dinitrate / Q000494:pharmacology*; D007700:Kinetics; D009569:Nitric Oxide / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000187:drug effects* / Q000378:metabolism; D005996:Nitroglycerin / Q000494:pharmacology; D010100:Oxygen / Q000494:pharmacology*; D010108:Oxyhemoglobins / Q000494:pharmacology; D013552:Swine
D010108:Oxyhemoglobins; D002956:Citrulline; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D005996:Nitroglycerin; D007548:Isosorbide Dinitrate; D010100:Oxygen