Dose-dependent relaxation of human venous vessel strips with regard to chronic nitrate pretreatment

Helical segments of human saphenous veins harvested at coronary bypass surgery were mounted in an organ-bath (Krebs-Henseleit buffer, pH 7.4; 37 degrees C; 95% O2/5% CO2 insufflation). After equilibration (60 min) and determination of basal tone the segments were depolarized and contracted by 24 mM potassium chloride. Then relaxation under isometric conditions was induced by cumulative concentrations of isosorbide dinitrate (ISDN) in a range of 10(-9) M to 10(-5) M. In vein segments from patients not pretreated with nitrates a concentration-response curve could be shown ranging from 8.4 +/- 4.5% to 71.9 +/- 8.6% (mean +/- SD). The relaxation pattern was not influenced by a 2-week pretreatment of patients with ISDN 20 mg twice daily or 40 mg four times daily even if the latter therapy was continued until 1 hour prior to surgery. Immersion of vessel strips in Krebs-Henseleit buffer containing 10(-6) M ISDN for 60 min prior to relaxation did not affect relaxation either. However, immersion of vessel segments in buffer medium containing 4.4 X 10(-4) M ISDN for 60 min led to a shift of the concentration-response curve by the factor 100 (EC50). Thus, chronic nitrate pretreatment of patients including high doses did not influence relaxation behaviour of isolated vessel segments. Induction of tolerance under in vitro conditions required concentrations exceeding the therapeutic limits. The most probable underlying mechanism is exhaustion of sulfhydril containing groups at the site of the smooth muscle cells. This hypothesis was supported by the finding that addition of cysteine into the organ-bath could widely reverse tolerance.(ABSTRACT TRUNCATED AT 250 WORDS)