Effect of Maturation on Endothelium-Dependent Relaxation in Pulmonary Arteries of the Newborn Rabbit

PURPOSE: This study was done to determine whether maturatin alters endothelium- dependent responses in pulmonary arteries. METHODS: Vascular rings of pulmonary arteries, with and without endothelium, taken from rabbits of 3 and 30 days of age were suspended in organ chambers filled with Krebs-Henseleit solution, bubbled with 95% O2-5% CO2 and maintained at 37degrees C. Immediately after mounting, the rings were stretched progressively until a maximal response to KCl was achieved. The rings were incubated with indomethacin and allowed to equilibrate before contraction and relaxation study. RESULTS: When the endothelium was intact in arterial rings from 3-day-old rabbits, acetylcholine (ACH) (10-6 M) relaxed preconstricted rings with histamine (5x10-6 M) (98.1 4.7% relaxation, mean SD). In rings without endothelium, KCl (10-2 to 9x10-2 M) and histamine (5x10-8 to 10-5 M) caused concentration-dependent contractions. When normalized to maximal contractions achieved to each agonist, the concentration-effect curves to KCl and histamine in rings without endothelium were similar to both ages. Rings with endothelium showed a progressive shift to the right of the concentration- effect curve to histamine. Relaxation to sodium nitroprusside were unaffected by age. In preconstricted ring, ACH (10-8 to 5x10-6 M) caused relaxations in rings with endothelium which were greater at 30-day compared to 3-day-old rabbits. CONCLUSION: These study demonstrates that endothelium-dependent relaxation increase with age, possibly due to changes in the release and/or effect of endothelium-derived relaxing factor (EDRF or nitric oxide) from pulmonary arteries during the neonatal period.

Phorbol Ester Modulates th Action of Acetylcholine in Rabbit Carotid Artery

Authors studied the regulatory mechanism of protein kinase C on the action of acetylcholine in rabbit carotid artery. The arterial rings were myographied isometrically in an isolated organ bath. In this study, acetylcholine relaxed phenylephrine-induced contraction of rabbit carotid artery in the presence of endothelium. In the pretreatment of methylene blue or nitro-L-arginine, the action of acetylchioline was reduced. Pretreatment of phorbol 12-myristate 13-acetate(PMA) attenuated the action of acetylcholine, but PMA did not attenuated it in the presence of staurosporine, suggesting that protein kinase C suppressed the action of acetylcholine. The potency of phorbol ester on the action of acetylcholine was PMA>phorbol 12, 13-dibutyrate(PDBu)>phorbol 12,13-diacetate(PDA), but the direct effect of phorbol on the contraction of arterial rings was PDBu>PMA>PDA. This implied that protein kinase C involved in the contraction of smooth muscle and the attenuation of the action of acetylcholine were different. PMA did not affect on A23187- and sodium nitroprusside-induced vasorelaxation. Acetylcholine increased tissue cGMP contents, which was reduced by PMA. These results suggest that in rabbit carotid artery protein kinase C reduce acetylcholine-stimuated endothelium derived relaxing factor(EDRF) release by affecting membrane receptor, and do not affect on the function of EDRF and cGMP production in the smooth muscle.