Cross-regulation of intracellular cGMP and cAMP in cultured human corpus cavernosum smooth muscle cells.
The goal of this study was to assess the potential cross-regulation of cyclic nucleotides in human corpus cavernosum (HCC). Incubation of primary cultures of HCC smooth muscle cells with either the NO donor sodium nitroprusside (SNP, 10 microM) or the phosphodiesterase type 5 (PDE 5) inhibitor sildenafil (50 nM) produced little or no changes in the intracellular cGMP levels. Incubation with both SNP and sildenafil produced marked increases in cGMP. Interestingly, incubation of cells with 10 microM of forskolin or PGE(1) produced significant enhancement of cGMP accumulation. These increases were not further enhanced by the addition of SNP and sildenafil. Kinetic analyses of cGMP hydrolysis by PDE 5 showed that high concentrations of cAMP reversibly inhibited the enzyme with a K(i) of 258 +/- 54 microM. The increase in cGMP levels in response to cAMP generating agents is not due to assay artifact since cAMP did not cross-react with cGMP antibody. Our data suggest that cAMP up-regulates intracellular levels of cGMP, in part, by inhibition of PDE 5. We also noted that cGMP down-regulates cAMP synthesis via a mechanism requiring G-protein coupling of adenylyl cyclase. These observations may have important implications in the utility of pharmacotherapeutic agents targeting cyclic nucleotide metabolism for the treatment of erectile dysfunction.
Kim
N N
NN
Department of Urology, Boston University School of Medicine, Boston, Massachusetts, 02118, USA.
Huang
Y
Y
Moreland
R B
RB
Kwak
S S
SS
Goldstein
I
I
Traish
A
A
D015106:3',5'-Cyclic-GMP Phosphodiesterases; D000262:Adenylyl Cyclases / Q000378:metabolism; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism*; D006152:Cyclic GMP / Q000378:metabolism*; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5; D004789:Enzyme Activation; D006801:Humans; D006868:Hydrolysis; D008297:Male; D009130:Muscle, Smooth / Q000166:cytology / Q000378:metabolism*; D010413:Penis / Q000166:cytology / Q000378:metabolism*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010727:Phosphoric Diester Hydrolases / Q000187:drug effects / Q000378:metabolism; D011494:Protein Kinases / Q000378:metabolism
D010726:Phosphodiesterase Inhibitors; D000242:Cyclic AMP; D011494:Protein Kinases; D010727:Phosphoric Diester Hydrolases; D015106:3',5'-Cyclic-GMP Phosphodiesterases; D054706:Cyclic Nucleotide Phosphodiesterases, Type 5; C517381:PDE5A protein, human; D000262:Adenylyl Cyclases; D006152:Cyclic GMP