Selective inhibition of cGMP-inhibited and cGMP-noninhibited cyclic nucleotide phosphodiesterases and relaxation of rat aorta.
In the supernatant (50,000 g, 1 hr) fraction from rat aortic smooth muscle homogenates, approximately 50% of total cAMPE PDE activity was inhibited by OPC 3911 (3 microM), while approximately 20% was inhibited by rolipram (30 microM). A cGMP-inhibited cyclic nucleotide phosphodiesterase (cGI-PDE) was further purified using DEAE chromatography followed by affinity chromatography on the N-(2-isothiocyanato)ethyl derivative of cilostamide conjugated to aminoethyl agarose (CIT-agarose). OPC 3911, CI-930, and milrinone, but not rolipram, were potent and selective inhibitors of this enzyme. The PDE-activity in the CIT-agarose flow through fraction (RI-PDE), however, was inhibited potently by rolipram, but not by cGMP, OPC 3911, CI-930 or milrinone. Functional studies showed that OPC 3911, CI-930, and milrinone were potent relaxants of contracted rat aorta. Rolipram had little relaxant effect. When OPC 3911 or milrinone was combined with rolipram more than additive effects on aortic relaxation and cAMP content were obtained. OPC 3911 combined with milrinone had only additive effects. These results demonstrate the presence of a cGI-PDE in rat aortic smooth muscle, and that inhibition of this isozyme may be of primary importance for the relaxant effects of OPC 3911, CI-930, and milrinone. A RI-PDE activity was also found, but it appeared to be less important for modulation of vascular tone unless the cGI-PDE was already inhibited. This may explain the synergistic relaxant effects observed when both PDE-isozymes were inhibited.
Lindgren
S
S
Department of Clinical Pharmacology, Lund University Hospital, Sweden.
Rascón
A
A
Andersson
K E
KE
Manganiello
V
V
Degerman
E
E
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D001011:Aorta; D000242:Cyclic AMP / Q000378:metabolism; D006152:Cyclic GMP / Q000494:pharmacology*; D004347:Drug Interactions; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D020105:Milrinone; D009119:Muscle Contraction / Q000187:drug effects; D009126:Muscle Relaxation / Q000187:drug effects; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000201:enzymology; D010656:Phenylephrine / Q000494:pharmacology; D011728:Pyridones / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology; D015363:Quinolones / Q000494:pharmacology; D051381:Rats; D020889:Rolipram; D012701:Serotonin / Q000494:pharmacology; D014665:Vasodilator Agents / Q000494:pharmacology*
D007527:Isoenzymes; D011728:Pyridones; D011760:Pyrrolidinones; D015363:Quinolones; D014665:Vasodilator Agents; D010656:Phenylephrine; D012701:Serotonin; C060853:OPC 3911; D000242:Cyclic AMP; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases; D006152:Cyclic GMP; D020105:Milrinone; D020889:Rolipram