Journal of Cardiovascular Pharmacology 2014-01-01

Potentiated adrenomedullin-induced vasorelaxation during hypoxia in organ cultured porcine coronary arteries.

Philip Hasbak, Majid Sheykhzade, Søren Schifter, Lars Edvinsson

Index: J. Cardiovasc. Pharmacol. 63(1) , 58-67, (2014)

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Abstract

This study describes the effect of variable oxygen supply on relaxing responses induced by α-calcitonin gene-related peptide (CGRP) and adrenomedullin (AM) on isolated pig coronary arteries in vitro. Organ culture during normoxia (21% of O₂) and hypoxia (5% of O₂) induced a significant leftward shift of the AM concentration-response curves compared with fresh vessels altering the pEC₅₀ values from 6.9 ± 0.04 to 8.0 ± 0.04, whereas the potency (pEC₅₀) of αCGRP was attenuated from 8.8 ± 0.04 to 7.6 ± 0.04. AM₂₂₋₅₂ exerted significant antagonistic effect on AM-induced vasorelaxation in hypoxic and normoxic conditions (apparent pK(B) = 6.8-7.2), whereas no antagonistic effect was observed in fresh and hyperoxic (95%) organ cultured vessels. The antagonistic effect exerted by αCGRP₈₋₃₇ (10⁻⁶·⁵-10⁻⁵·⁵ M) on αCGRP-induced vasodilatation in fresh vessels (derived from Schild plot pA₂ = 7.4 ± 0.1) was unaltered during organ culture. The antagonistic effect exerted by αCGRP₈₋₃₇ (10⁻⁶ M) on AM-induced vasorelaxation in fresh vessels (apparent pK(B) = 7.4 ± 0.1) was absent during hypoxic organ culture. The receptor activity-modifying proteins 1 (RAMP1)/calcitonin-like receptor (CLR) messenger RNA ratio was reduced and RAMP2/CLR messenger RNA ratio was increased during hypoxic and normoxic organ culture compared with fresh vessels. Hypoxic organ culture for 24-72 hours potentiated the AM-induced vasorelaxation through an AM₂₂₋₅₂-sensitive receptor but attenuated the vasorelaxant effect of CGRP through the CGRP receptors. This could possibly be explained by relatively decreased levels of RAMP1, thus favoring RAMP2 + CLR complex (=AM receptor) formation during hypoxic organ culture.