Role of Platelet-Activating Factor and Hypoxia in Persistent Pulmonary Hypertension of the Newborn — Studies with Perinatal Pulmonary Vascular Smooth Muscle Cells

Abstract

Platelet-activating factor (PAF) plays an important physiological role of maintaining a high vasomotor tone in fetal pulmonary circulation. At birth, endogenous vasodi‐ lators such as nitric oxide and prostacyclin are released and facilitate pulmonary vasodilation via cAMP-dependent protein kinase (cAMP/PKA) and cGMP-dependent protein kinase (cGMP/PKG) pathways. Interaction between the cyclic nucleotides and PAF receptor (PAFR)-mediated responses in pulmonary arterial smooth muscle is not well understood. To further understand the interactions of PAF-PAFR pathway and the cyclic nucleotides in ovine fetal pulmonary arterial smooth muscle cells (FPASMC), effects of cAMP and cGMP on PAFR-mediated responses in pulmonary arterial smooth muscle cells (PASMC) were studied. Ovine FPASMC were incubated with 10μM cAMP or cGMP in normoxia (5% CO2 in air, pO2~100 Torr) or hypoxia (2% O2, 5% CO2, pO2~30-40 Torr). Proteins were prepared and subjected to Western blotting. Effect of cell permeable cAMP and cGMP on PAFR binding was also studied and effect of cAMP on cell proliferation was also studied by RNAi to PKA-Cα. cAMP and cGMP significantly decreased PAFR binding and protein expression in normoxia and hypoxia, more so in hypoxia, when PAFR expression was usually high. PKACα siRNA demonstrated that inhibition of PAFR-mediated responses by the cyclic nucleotides occurred through PKA. These data suggest that the normally high levels © 2015 The Author(s). Licensee InTech. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. of cyclic nucleotides in the normoxic newborn pulmonary circulation assist in the downregulation of postnatal PAFR-mediated responses and that under hypoxic conditions, increasing the levels of cyclic nucleotides will abrogate PAF-mediated vasoconstriction thereby ameliorating PAF-induced persistent pulmonary hyperten‐ sion of the newborn.