nitroarginine and 8-tert-butyl-6-7-dihydropyrrolo(3-2-e)-5-methylpyrazolo(1-5-a)pyrimidine-3-carbonitrile

1. LP-805, 8-tert-butyl-6,7-dihydropyrrolo-[3,2-e]-5-methylpyrazolo- [1,5a]-pyrimidine-3-carbonitrile, is a newly synthesized potent vasodilator. To investigate the cellular mechanisms of vasorelaxation induced by LP-805, we simultaneously determined the effects of LP-805 on cytosolic Ca2+ concentrations ([Ca2+]i) and on tension of smooth muscle of rabbit femoral arterial strips, with or without the endothelium, using front-surface fluorometry and fura-2. 2. In the absence of the endothelium, LP-805, in a concentration-dependent manner, decreased [Ca2+]i and tension during the contraction induced by K(+)-depolarization, at relatively low concentrations ([K+]o < or = 30 mM). The decreases in [Ca2+]i and tension were fully antagonized by treatment with 2 x 10(-6) M glibenclamide. The [Ca2+]i-tension relationship in the LP-805-induced relaxation was similar to that of K(+)-depolarization-induced contractions. 3. LP-805, in a concentration-dependent manner (IC50 for inhibition of tension; 1.7 x 10(-6) M), decreased both [Ca2+]i and tension during the steady-state of contractions induced by 1 x 10(-7) M noradrenaline (NA) in the strips without the endothelium. Glibenclamide completely inhibited these reductions of [Ca2+]i and tension. At the steady-state of relaxation induced by LP-805 during NA-induced contraction, [Ca2+]i-tension relation was shifted to the left of that obtained with high K(+)-induced contraction. 4. NA induced transient increases in [Ca2+]i and tension in the absence of extracellular Ca2+. LP-805 (up to 3 x 10(-6) M) had no effect on these intracellular Ca2+ mobilisation and tension development induced by NA. 5. In strips with an intact endothelium, LP-805 decreased both [Ca2+]i and tension during contraction induced by 1 x 10(-7) M NA. The concentration-response curve for inhibition of [Ca2+]i and tension obtained in the presence of the endothelium was shifted to the left from that obtained in the absence of endothelium. IC50 for the inhibition of tension obtained in the strips with the endothelium was 4.0 x 10(-7) M. Treatment with 1 x 10(-4) M NG-nitro-L-arginine (L-NOARG) attenuated reductions of both [Ca2+]i and tension induced by LP-805 and the concentration-response curve shifted to the right and overlapped that obtained in the absence of the endothelium. Treatment with glibenclamide almost fully overcame the reduction of [Ca2+]i induced by LP-805, while the reversion of tension was 50% at most. 6. In the presence of the endothelium

Topics: Animals; Arginine; Benzopyrans; Calcium; Cromakalim; Cytosol; Endothelium, Vascular; Femoral Artery; Fura-2; In Vitro Techniques; Male; Muscle Contraction; Muscle, Smooth, Vascular; Nitric Oxide; Nitroarginine; Norepinephrine; Potassium; Pyrazoles; Pyrimidines; Pyrroles; Rabbits; Vasodilator Agents

We have investigated relations between hypotensive responses to LP-805, a newly synthesized vasodilator, and the production of nitric oxide (NO), in anesthetized rats. LP-805 (0.1-0.5 mg/kg, i.v.) or acetylcholine (ACh) (0.3-3.0 micrograms/kg, i.v.) caused a dose-dependent transient decrease in diastolic blood pressure. The decrease induced by 0.3 mg/kg LP-805 (i.v.) was partially inhibited by pretreatment with NG-nitro-L-arginine (L-NNA), a specific inhibitor of endothelial NO synthase, but the responses to lower or higher doses of LP-805 (0.1 or 0.5 mg/kg, i.v.) were not affected. The dose-dependent decrease in diastolic blood pressure, caused by LP-805, was not affected by pretreatment with L- or D-arginine. The dose-dependent decrease in diastolic blood pressure caused by ACh was not affected by pretreatment with L-NNA or with L- or D-arginine. The hypotensive response to 20-min infusions of LP-805 (100 micrograms/kg per min) was significantly inhibited by pretreatment with L-NNA (10 mg/kg, i.v.). The half-recovery times (T 1/2) of LP-805 or ACh-induced depressor responses were shortened by pretreatment with L-NNA. They were prolonged by L-arginine, but not by D-arginine. This shortening, by L-NNA, of the half-recovery time after LP-805 or ACh was reversed by L-arginine, but not by D-arginine. The T 1/2 of the LP-805-induced hypotensive response was not affected by pretreatment with indomethacin (1 mg/kg, i.v.). In the presence of L-NNA (10 mg/kg, i.v.), the T 1/2 of the LP-805-induced hypotensive response was not affected by pretreatment with indomethacin.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acetylcholine; Animals; Arginine; Blood Pressure; Dose-Response Relationship, Drug; Endothelium, Vascular; Hypotension; Indomethacin; Injections, Intravenous; Male; Nitric Oxide; Nitroarginine; Pyrazoles; Pyrimidines; Rats; Rats, Wistar; Vasodilation; Vasodilator Agents