Involvement of protein kinase C in reduced relaxant responses to the NO/cyclic GMP pathway in piglet pulmonary arteries contracted by the thromboxane A2-mimetic U46619

Abstract

1. Impairment of nitric oxide (NO)/cyclic GMP production and/or increased activities of thromboxane A2 (TXA2) and endothelin-1 (ET-1) have been associated with pulmonary hypertension. We have analysed the interactions of noradrenaline (NA), the TXA2-mimetic U46619 and ET-1 with the relaxation induced via cyclic GMP in isolated piglet intrapulmonary arteries. 2. The contractions induced by NA were augmented by endothelium removal or by methylene blue and pre-contracted rings were fully relaxed by acetylcholine, sodium nitroprusside (SNP), atrial natriuretic peptide and 8-bromo-cyclic GMP. In contrast, U46619- and ET-1 induced contractions were endothelium-independent and only partially relaxed by the latter vasodilators. Whereas the reduced responses to SNP in arteries contracted by U46619 were independent of the U46619-induced tone, a higher concentration of ET-1 (tone higher than that induced by NA) was required to reduce the vasodilator responses to SNP. NA, U46619 and ET-1 had no effect on the SNP-induced increases in cyclic GMP. 3. The reduced relaxant responses to SNP in arteries pre-contracted by U46619 were specific for piglet pulmonary arteries since they were not observed in piglet mesenteric or coronary arteries or in rat pulmonary arteries. Furthermore, there were no differences in the relaxant response to the adenylate cyclase activator forskolin in piglet pulmonary arteries pre-contracted by either NA, U46619 or ET-1. 4. SNP-induced relaxation was inhibited by thapsigargin (but not by inhibition of the membrane Na+/ K+ ATPase nor K+ channels) indicating a role for Ca2+ sequestration by the Ca2+ ATPase in the effects of SNP. 5. The phorbol ester 12-myristate, 13-acetate inhibited the relaxant response to SNP. The inhibitory effect of U46619 on SNP-induced relaxation was abolished by the protein kinase C inhibitor (PKC) staurosporine suggesting that PKC may be a part of the signal transduction mechanism. 6. In summary, piglet pulmonary arteries when activated by a TXA2-mimetic show abnormally reduced relaxant responses to the NO/cyclicGMP pathway. This effect appears to be mediated by activation of PKC.