Nitric oxide induces phosphodiesterase 4B expression in rat pulmonary artery smooth muscle cells

1 Department of Anesthesia and Critical Care and 2 Cardiovascular Research Center of the Department of Medicine, Massachusetts General Hospital, Harvard Medical School, Charlestown, Massachusetts Submitted 11 July 2005 ; accepted in final form 17 November 2005 Phosphodiesterases (PDE) metabolize cyclic nucleotides limiting the effects of vasodilators such as prostacyclin and nitric oxide (NO). In this study, DNA microarray techniques were used to assess the impact of NO on expression of PDE genes in rat pulmonary arterial smooth muscle cells (rPASMC). Incubation of rPASMC with S -nitroso- L -glutathione (GSNO) increased expression of a PDE isoform that specifically metabolizes cAMP (PDE4B) in a dose- and time-dependent manner. GSNO increased PDE4B protein levels, and rolipram-inhibitable PDE activity was 2.3 ± 1.0-fold greater in GSNO-treated rPASMC than in untreated cells. The soluble guanylate cyclase (sGC) inhibitor, 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one, and the cAMP-dependent protein kinase inhibitor, H89, prevented induction of PDE4B gene expression by GSNO, but the protein kinase G (PKG) inhibitors, Rp-8-pCPT-cGMPs and KT-5823, did not. Incubation of rPASMC with IL-1 and tumor necrosis factor- induced PDE4B gene expression, an effect that was inhibited by L - N 6 -(1-iminoethyl)lysine, an antagonist of NO synthase 2 (NOS2). The GSNO-induced increase in PDE4B mRNA levels was blocked by actinomycin D but augmented by cycloheximide. Infection of rPASMC with an adenovirus specifying a dominant negative cAMP response element binding protein (CREB) mutant inhibited the GSNO-induced increase of PDE4B gene expression. These results suggest that exposure of rPASMC to NO induces expression of PDE4B via a mechanism that requires cGMP synthesis by sGC but not PKG. The GSNO-induced increase of PDE4B gene expression is CREB dependent. These findings demonstrate that NO increases expression of a cAMP-specific PDE and provide evidence for a novel "cross talk" mechanism between cGMP and cAMP signaling pathways. cross talk; S -nitroso- L -glutathione; adenosine 3',5'-cyclic monophosphate; guanosine 3',5'-cyclic monophosphate Address for reprint requests and other correspondence: C. J. Busch, Dept. of Anesthesiology, Ruprecht Karl University, Im Neuenheimer Feld 110, 69120 Heidelberg, Germany (e-mail: Cornelius_Busch{at}med.uni-heidelberg.de )