Differential effects of phosphodiesterase inhibitors on accumulation of cyclic AMP in isolated ventricular cardiomyocytes.
The intracellular actions of phosphodiesterase (PDE) inhibitors on the accumulation of cyclic nucleotides were studied in isolated ventricular cardiomyocytes from adult Sprague-Dawley rats. Elevated levels of cyclic AMP, due to the effects of selective PDE inhibitors, were detected only when the levels of cyclic nucleotide were enhanced with forskolin (10 microM). The time course for the elevation of cyclic AMP levels was similar for all the PDE inhibitors tested, following the pattern of an initial rise in the first 2-4 min, proceeded by a steady state at 67 +/- 6% of the maximum stimulation. HN-10200 (2-[3-methoxy-5-methylsulfinyl-2-thienyl]-1H-imidazo-[4,5-c]- pyridine hydrochloride), a new imidazopyridine derivative, had a similar concentration-dependent profile to the structurally related compound, sulmazole (AR-L 115 BS, 2-[2-methoxy-4-methylsulfinyl)phenyl]-1H- imidazo-[4,5-b]-pyridine). Both the non-selective inhibitor, 3-isobutyl-1-methylxanthine (IBMX), and the selective PDE IV inhibitor, Ro 20-1724 (4-[(3-butoxy-4-methoxyphenyl)methyl]-2- imidazolidinone), potentiated the forskolin-stimulated levels of cyclic AMP with a much greater efficacy than sulmazole or HN-10200. The concentrations of forskolin required by IBMX, sulmazole and HN-10200 (10(-3) M) to increase levels of cyclic AMP by 4 pmol/mg protein were 3.2 x 10(-6) M, 1.32 x 10(-5) M and 1.46 x 10(-5) M, respectively. Enoximone failed to cause an increase in the levels of cyclic AMP, even when stimulated with maximal concentrations of forskolin. Furthermore, in the presence of forskolin, enoximone attenuated the response of Ro 20-1724 and IBMX in a concentration-dependent manner. Enoximone, similarly to HN-10200, sulmazole, Ro 20-1724 and IBMX did not produce any significant effect on levels of cyclic GMP under elevated conditions in the presence of sodium nitroprusside. The combined action of Ro 20-1724, with either HN-10200, sulmazole, or IBMX (10(-4) M), on intracellular levels of cyclic AMP, was not greater than the response to Ro 20-1724 alone. These data demonstrate the differential actions of PDE III and PDE IV inhibitors in rat ventricular cardiomyocytes. It is suggested that enoximone has a high selectivity for the PDE III isoenzyme so that hydrolysis of cyclic AMP by the PDE IV isoenzyme is not inhibited, in accordance with the lack of increase in cyclic AMP by enoximone in rat cardiomyocytes. HN-10200 and sulmazole, producing small increases in intracellular levels of cyclic AMP, are less selective PDE III inhibitors than enoximone.(ABSTRACT TRUNCATED AT 400 WORDS)
Kelso
E J
EJ
Department of Therapeutics and Pharmacology, Queen's University of Belfast, U.K.
McDermott
B J
BJ
Silke
B
B
D000818:Animals; D002478:Cells, Cultured; D005576:Colforsin; D000242:Cyclic AMP / Q000378:metabolism*; D017335:Enoximone / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D007527:Isoenzymes / Q000378:metabolism; D007545:Isoproterenol; D008297:Male; D009206:Myocardium / Q000378:metabolism*; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley
C077127:HN 10200; D007093:Imidazoles; D007527:Isoenzymes; D010726:Phosphodiesterase Inhibitors; D011725:Pyridines; D005576:Colforsin; D017335:Enoximone; D000242:Cyclic AMP; D010727:Phosphoric Diester Hydrolases; C029758:sulmazole; D007545:Isoproterenol