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Claire Lugnier - One of the best experts on this subject based on the ideXlab platform.
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Disease Progression in MRL/lpr Lupus-Prone Mice Is Reduced by NCS 613, a Specific Cyclic Nucleotide Phosphodiesterase Type 4 (PDE4) Inhibitor
2016Co-Authors: Fanny Monneaux, Lucien Gazi, Jean-jacques Bourguignon, Sylviane Muller, Claire LugnierAbstract:Systemic lupus erythematosus is a polymorphic and multigenic inflammatory autoimmune disease. Cyclic AMP (cAMP) modulates inflammation and the inhibition of Cyclic Nucleotide Phosphodiesterase type 4 (PDE4), which specifically hydrolyzes cAMP, inhibits TNFa secretion. This study was aimed at investigating the evolution of PDE activity and expression levels during the course of the disease in MRL/lpr lupus-prone mice, and to evaluate in these mice the biological and clinical effects of treatments with pentoxifylline, denbufylline and NCS 613 PDE inhibitors. This study reveals that compared to CBA/ J control mice, kidney PDE4 activity of MRL/lpr mice increases with the disease progression. Furthermore, it showed that the most potent and selective PDE4 inhibitor NCS 613 is also the most effective molecule in decreasing proteinuria and increasing survival rate of MRL/lpr mice. NCS 613 is a potent inhibitor, which is more selective for the PDE4C subtyp
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Disease progression in MRL/lpr lupus-prone mice is reduced by NCS 613, a specific Cyclic Nucleotide Phosphodiesterase type 4 (PDE4) inhibitor.
PloS one, 2012Co-Authors: Thérèse Keravis, Issaka Yougbaré, Fanny Monneaux, Lucien Gazi, Jean-jacques Bourguignon, Sylviane Muller, Claire LugnierAbstract:Systemic lupus erythematosus is a polymorphic and multigenic inflammatory autoimmune disease. Cyclic AMP (cAMP) modulates inflammation and the inhibition of Cyclic Nucleotide Phosphodiesterase type 4 (PDE4), which specifically hydrolyzes cAMP, inhibits TNFα secretion. This study was aimed at investigating the evolution of PDE activity and expression levels during the course of the disease in MRL/lpr lupus-prone mice, and to evaluate in these mice the biological and clinical effects of treatments with pentoxifylline, denbufylline and NCS 613 PDE inhibitors. This study reveals that compared to CBA/J control mice, kidney PDE4 activity of MRL/lpr mice increases with the disease progression. Furthermore, it showed that the most potent and selective PDE4 inhibitor NCS 613 is also the most effective molecule in decreasing proteinuria and increasing survival rate of MRL/lpr mice. NCS 613 is a potent inhibitor, which is more selective for the PDE4C subtype (IC50 = 1.4 nM) than the other subtypes (PDE4A, IC50 = 44 nM; PDE4B, IC50 = 48 nM; and PDE4D, IC50 = 14 nM). Interestingly, its affinity for the High Affinity Rolipram Binding Site is relatively low (Ki = 148 nM) in comparison to rolipram (Ki = 3 nM). Finally, as also observed using MRL/lpr peripheral blood lymphocytes (PBLs), NCS 613 inhibits basal and LPS-induced TNFα secretion from PBLs of lupus patients, suggesting a therapeutic potential of NCS 613 in systemic lupus. This study reveals that PDE4 represent a potential therapeutic target in lupus disease.
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Cyclic Nucleotide Phosphodiesterase pde superfamily a new target for the development of specific therapeutic agents
Pharmacology & Therapeutics, 2006Co-Authors: Claire LugnierAbstract:Cyclic Nucleotide Phosphodiesterases (PDEs), which are ubiquitously distributed in mammalian tissues, play a major role in cell signaling by hydrolyzing cAMP and cGMP. Due to their diversity, which allows specific distribution at cellular and subcellular levels, PDEs can selectively regulate various cellular functions. Their critical role in intracellular signaling has recently designated them as new therapeutic targets for inflammation. The PDE superfamily represents 11 gene families (PDE1 to PDE11). Each family encompasses 1 to 4 distinct genes, to give more than 20 genes in mammals encoding the more than 50 different PDE proteins probably produced in mammalian cells. Although PDE1 to PDE6 were the first well-characterized isoforms because of their predominance in various tissues and cells, their specific contribution to tissue function and their regulation in pathophysiology remain open research fields. This concerns particularly the newly discovered families, PDE7 to PDE11, for which roles are not yet established. In many pathologies, such as inflammation, neurodegeneration, and cancer, alterations in intracellular signaling related to PDE deregulation may explain the difficulties observed in the prevention and treatment of these pathologies. By inhibiting specifically the up-regulated PDE isozyme(s) with newly synthesized potent and isozyme-selective PDE inhibitors, it may be potentially possible to restore normal intracellular signaling selectively, providing therapy with reduced adverse effects.
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implication of Cyclic Nucleotide Phosphodiesterase inhibition in the vasorelaxant activity of the citrus fruits flavonoid naringenin
Planta Medica, 2005Co-Authors: Francisco Orallo, Mercedes Camina, Ezequiel Alvarez, Helene Basaran, Claire LugnierAbstract:The potential vasorelaxant, antioxidant and Cyclic Nucleotide Phosphodiesterase (PDE) inhibitory effects of the citrus-fruit flavonoids naringin and (+/-)-naringenin were comparatively studied for the first time in this work. (+/-)-Naringenin (1 microM - 0.3 mM) did not affect the contractile response induced by okadaic acid (OA, 1 microM). However, (+/-)-naringenin relaxed, in a concentration-dependent manner, the contractions elicited by phenylephrine (PHE, 1 microM) or by a high extracellular KCl concentration (60 mM) in intact rat aortic rings. Mechanical removal of endothelium and/or pretreatment of aorta rings with glibenclamide (GB, 10 microM) or tetraethylammonium (TEA, 2 mM) did not significantly modify the vasorelaxant effects of this flavanone. (+/-)-Naringenin (10 microM - 0.1 mM) did not alter the basal uptake of 4) Ca2+ but decreased the influx of 45Ca2+ induced by PHE and KCl in endothelium-containing and endothelium-denuded rat aorta. (+/-)-Naringenin (10 microM - 0.1 mM) was ineffective to scavenge superoxide radicals (O*2-) generated by the hypoxanthine (HX)-xanthine oxidase (XO) system and/or to inhibit XO activity. (+/-)-Naringenin (0.1 mM) significantly increased the production of cGMP and cAMP decreased by PHE (1 microM) and high KCl (60 mM) in cultured rat aortic myocytes. (+/-)-Naringenin preferentially inhibited calmodulin (CaM)-activated PDE1, PDE4 and PDE5 isolated from bovine aorta with IC50 values of about 45 microM, 60 microM and 68 microM, respectively. In contrast, the 7-rhamnoglucoside of (+/-)-naringenin, naringin (1 microM - 0.3 mM), was totally inactive in all experiments. These results indicate that the vasorelaxant effects of (+/-)-naringenin seem to be basically related to the inhibition of PDE1, PDE4 and PDE5 activities.
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effect of sildenafil on Cyclic Nucleotide Phosphodiesterase activity vascular tone and calcium signaling in rat pulmonary artery
British Journal of Pharmacology, 2003Co-Authors: O Pauvert, Claire Lugnier, Thérèse Keravis, Roger Marthan, Eric Rousseau, Jeanpierre SavineauAbstract:Sildenafil (viagra) is a potent PDE5 inhibitor and thus a relaxant drug in corpus carvernosum smooth muscle. In the present work, we evidenced the presence of PDE5 isozyme and investigated the effect of sildenafil on the specific Cyclic Nucleotide Phosphodiesterase (PDE) activity, smooth muscle tone and calcium signaling in the rat main pulmonary artery (MPA). The PDE activity was measured in cytosolic and microsomal fractions. Total cAMP and cGMP-PDE activities were mainly present in the cytosolic fraction. Sildenafil (0.1 μM) reduced by 72% cGMP-PDE activity, whereas zaprinast (10 μM), a relatively selective PDE5 inhibitor, reduced this activity by 63%. Sildenafil (0.1 μM) also inhibited significantly (22%) the cAMP-PDE activity. Western blot analysis revealed the expression of PDE5 mainly in the cytosolic fraction of MPA. Sildenafil concentration-dependently inhibited (IC50=3.4 nM) the activity of MPA PDE5 partially purified by HPLC. Sildenafil (0.1 nM–50 μM) concentration-dependently relaxed MPA rings precontracted with phenylephrine (0.5 μM). The potency of sildenafil (IC50=11 nM) was similar to that of a nitric oxide donor, sodium nitroprusside, but higher than that of zaprinast (IC50=600 nM). The vasorelaxant effect of sildenafil was not altered by endothelium removal or in the presence of KT 5823 (1 μM) and H89 (1 μM), potent inhibitors of PKG and PKA, respectively. In isolated MPA myocytes, which had been loaded with the calcium fluorophore indo-1, sildenafil (10–100 nM) antagonized ATP- and endothelin-1-induced calcium oscillations but had no effect on the transient caffeine-induced [Ca2+]i response. This study demonstrates the presence of a functional and highly sildenafil-sensitive PDE5 isozyme in rat MPA. Inhibition of this isozyme mainly accounts for the potent pulmonary vasodilator action of sildenafil, which involves alteration in the inositol triphosphate-mediated calcium signaling pathway. Keywords: Sildenafil, Cyclic Nucleotide Phosphodiesterase, Phosphodiesterase inhibitor, Pulmonary artery, Ca2+ signaling, Vascular tone Introduction Cyclic Nucleotides (cAMP and cGMP) are involved in the control of smooth muscle tone, as an increase in their intracellular concentration generally leads to relaxation mainly via an alteration in calcium signaling (Polson & Strada, 1996; Carvajal et al., 2000). Cyclic Nucleotide Phosphodiesterases (PDEs), which hydrolyze cAMP and cGMP, represent the unique degradation pathway for these intracellular compounds (Stoclet et al., 1995). As a consequence, PDE activity is also implicated in the control of smooth muscle tone and PDE inhibition relaxes smooth muscle (Schoeffter et al., 1987; Komas et al., 1991; Wagner et al., 1997). Mammalian cells contain a variety of PDEs that have been classified into 11 gene families based on their primary structures, and their catalytic and regulatory properties (Soderling & Beavo, 2000; Francis et al., 2001). In mammalian smooth muscle, at least, five PDE isozymes (PDE1–PDE5) have been identified. Sildenafil citrate (Viagra™) has been shown as a potent (IC50≅1–4 nM) and reversible inhibitor of PDE5 (Moreland et al., 1998; Illarion et al., 1999). Moreover, it is highly selective for this PDE when compared with other PDE families (Ballard et al., 1998). In the corpus cavernosum, sildenafil increases cGMP concentration and causes smooth muscle relaxation of arteries and sinusoids, thus facilitating blood flow (Jeremy et al., 1997; Ballard et al., 1998). As a consequence, sildenafil has proved to be an effective, well-tolerated drug in the treatment of erectile dysfunction (Boolell et al., 1996; Ballard et al., 1998). Pulmonary circulation develops a specific response to hypoxia, that is, vasoconstriction which can be maintained in patients suffering from pulmonary obstructive diseases (e.g., chronic bronchitis) (Rounds, 1989; Pierson, 2000), thus inducing, in turn, a sustained elevation in the pulmonary blood pressure that causes pulmonary hypertension (PAHT). PAHT leads to right ventricular hypertrophy, right heart failure and ultimately to death (Rabinovitch et al., 1979; Dawson, 1984). This pathophysiological adaptation of the pulmonary circulation to maintain hypoxia is a complicated process for which no curative treatment, except heart–lung transplantation, is currently available. Since PDEs are present in the pulmonary artery wall (Rabe et al., 1994; Maclean et al., 1997) and that cGMP-PDE activity is mainly because of the action of PDE5 (Rabe et al., 1994), some PDE5 inhibitors have been administered to counteract PAHT development both in animals and human (Eddahibi et al., 1998; Ziegler et al., 1998; Hanasato et al., 1999). Very recently, pioneer clinical trials with sildenafil have been performed in PAHT (Abrams et al., 2000; Ichinose et al., 2001; Zhao et al., 2001). However, to the best of our knowledge, no detailed study has been performed, at the cellular and molecular levels, to examine the interactions of sildenafil and cGMP-PDE signaling pathway in the pulmonary vascular smooth muscle. Thus, the aims of the present study conducted in the rat pulmonary artery were: (1) to measure PDE5-specific activity in comparison with other cAMP- and cGMP-PDE activities both in cytosolic and microsomal fractions; (2) to examine the expression of PDE5 protein in the arterial wall; (3) to quantify the effect of sildenafil on the partially purified PDE5; (4) to determine the relaxant properties of sildenafil on pulmonary tone and its effect at the site of calcium signaling in vascular smooth muscle cells.
W J Thompson - One of the best experts on this subject based on the ideXlab platform.
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erythro 9 2 hydroxy 3 nonyl adenine inhibits Cyclic 3 5 guanosine monophosphate stimulated Phosphodiesterase to reverse hypoxic pulmonary vasoconstriction in the perfused rat lung
Journal of Pharmacology and Experimental Therapeutics, 1996Co-Authors: J Haynes, D W Killilea, P D Peterson, W J ThompsonAbstract:Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was shown to reverse the hypoxic pressor response (HPR) in the isolated, blood-perfused rat lung model. EHNA, an adenosine deaminase inhibitor, showed reversal of the HPR in a dose-dependent manner (EC50 = 129 +/- 30 microM). We found that the reversal of HPR by EHNA was not mediated by the adenosine receptors because the EHNA effect was not blocked by the adenosine receptor antagonist, 8-p-sulfophenyl-theophylline (67 microM; n = 6). Pretreatment with a cy-clic-39,59-adenosine monophosphate (cAMP)-dependent protein kinase inhibitor, Rp-adenosine-39,59-Cyclic monophosphorothioate (0.5 mM; n = 4), blocked EHNA reversal of the HPR. As an alternative mechanism of action, EHNA inhibition of Cyclic Nucleotide Phosphodiesterase(s) isozymes was studied in endothelium intact and denuded pulmonary arteries. Using anion-exchange chromatography the Cyclic Nucleotide Phosphodiesterase (PDE) separated into predominantly PDE families 2 and a mixture of 3 and 4. DEAE fractions showing cAMP hydrolysis activated by 5 microM Cyclic-39,59-guanosine monophosphate (cGMP) had a Km for cAMP of 6.3 microM and an apparent Kact for cGMP of 1.4 microM. EHNA was shown to inhibit PDE2 competitively. In intact vessels, the IC50 for EHNA was 3.3 microM using 0.03 microM [3H]-cAMP substrate assayed in the presence of 2 microM cGMP and in denuded vessels 3.7 microM at 0.03 microM [3H]-cAMP substrate in the presence of 5 microM cGMP. Fractions in which cAMP hydrolysis was inhibited or not affected by 5 microM cGMP (PDE3 and 4, respectively) showed an IC50 of > 200 microM for EHNA. We conclude that reversal of the hypoxic pressor response by EHNA in the isolated, perfused rat lung model occurs with a mechanism involving in part inhibition of smooth muscle PDE2.
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erythro 9 2 hydroxy 3 nonyl adenine inhibits Cyclic 3 5 guanosine monophosphate stimulated Phosphodiesterase to reverse hypoxic pulmonary vasoconstriction in the perfused rat lung
Journal of Pharmacology and Experimental Therapeutics, 1996Co-Authors: J Haynes, D W Killilea, P D Peterson, W J ThompsonAbstract:Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was shown to reverse the hypoxic pressor response (HPR) in the isolated, blood-perfused rat lung model. EHNA, an adenosine deaminase inhibitor, showed reversal of the HPR in a dose-dependent manner (EC50 = 129 +/- 30 microM). We found that the reversal of HPR by EHNA was not mediated by the adenosine receptors because the EHNA effect was not blocked by the adenosine receptor antagonist, 8-p-sulfophenyl-theophylline (67 microM; n = 6). Pretreatment with a cy-clic-3',5'-adenosine monophosphate (cAMP)-dependent protein kinase inhibitor, Rp-adenosine-3',5'-Cyclic monophosphorothioate (0.5 mM; n = 4), blocked EHNA reversal of the HPR. As an alternative mechanism of action, EHNA inhibition of Cyclic Nucleotide Phosphodiesterase(s) isozymes was studied in endothelium intact and denuded pulmonary arteries. Using anion-exchange chromatography the Cyclic Nucleotide Phosphodiesterase (PDE) separated into predominantly PDE families 2 and a mixture of 3 and 4. DEAE fractions showing cAMP hydrolysis activated by 5 microM Cyclic-3',5'-guanosine monophosphate (cGMP) had a Km for cAMP of 6.3 microM and an apparent Kact for cGMP of 1.4 microM. EHNA was shown to inhibit PDE2 competitively. In intact vessels, the IC50 for EHNA was 3.3 microM using 0.03 microM [3H]-cAMP substrate assayed in the presence of 2 microM cGMP and in denuded vessels 3.7 microM at 0.03 microM [3H]-cAMP substrate in the presence of 5 microM cGMP. Fractions in which cAMP hydrolysis was inhibited or not affected by 5 microM cGMP (PDE3 and 4, respectively) showed an IC50 of > 200 microM for EHNA. We conclude that reversal of the hypoxic pressor response by EHNA in the isolated, perfused rat lung model occurs with a mechanism involving in part inhibition of smooth muscle PDE2.
J Haynes - One of the best experts on this subject based on the ideXlab platform.
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erythro 9 2 hydroxy 3 nonyl adenine inhibits Cyclic 3 5 guanosine monophosphate stimulated Phosphodiesterase to reverse hypoxic pulmonary vasoconstriction in the perfused rat lung
Journal of Pharmacology and Experimental Therapeutics, 1996Co-Authors: J Haynes, D W Killilea, P D Peterson, W J ThompsonAbstract:Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was shown to reverse the hypoxic pressor response (HPR) in the isolated, blood-perfused rat lung model. EHNA, an adenosine deaminase inhibitor, showed reversal of the HPR in a dose-dependent manner (EC50 = 129 +/- 30 microM). We found that the reversal of HPR by EHNA was not mediated by the adenosine receptors because the EHNA effect was not blocked by the adenosine receptor antagonist, 8-p-sulfophenyl-theophylline (67 microM; n = 6). Pretreatment with a cy-clic-39,59-adenosine monophosphate (cAMP)-dependent protein kinase inhibitor, Rp-adenosine-39,59-Cyclic monophosphorothioate (0.5 mM; n = 4), blocked EHNA reversal of the HPR. As an alternative mechanism of action, EHNA inhibition of Cyclic Nucleotide Phosphodiesterase(s) isozymes was studied in endothelium intact and denuded pulmonary arteries. Using anion-exchange chromatography the Cyclic Nucleotide Phosphodiesterase (PDE) separated into predominantly PDE families 2 and a mixture of 3 and 4. DEAE fractions showing cAMP hydrolysis activated by 5 microM Cyclic-39,59-guanosine monophosphate (cGMP) had a Km for cAMP of 6.3 microM and an apparent Kact for cGMP of 1.4 microM. EHNA was shown to inhibit PDE2 competitively. In intact vessels, the IC50 for EHNA was 3.3 microM using 0.03 microM [3H]-cAMP substrate assayed in the presence of 2 microM cGMP and in denuded vessels 3.7 microM at 0.03 microM [3H]-cAMP substrate in the presence of 5 microM cGMP. Fractions in which cAMP hydrolysis was inhibited or not affected by 5 microM cGMP (PDE3 and 4, respectively) showed an IC50 of > 200 microM for EHNA. We conclude that reversal of the hypoxic pressor response by EHNA in the isolated, perfused rat lung model occurs with a mechanism involving in part inhibition of smooth muscle PDE2.
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erythro 9 2 hydroxy 3 nonyl adenine inhibits Cyclic 3 5 guanosine monophosphate stimulated Phosphodiesterase to reverse hypoxic pulmonary vasoconstriction in the perfused rat lung
Journal of Pharmacology and Experimental Therapeutics, 1996Co-Authors: J Haynes, D W Killilea, P D Peterson, W J ThompsonAbstract:Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was shown to reverse the hypoxic pressor response (HPR) in the isolated, blood-perfused rat lung model. EHNA, an adenosine deaminase inhibitor, showed reversal of the HPR in a dose-dependent manner (EC50 = 129 +/- 30 microM). We found that the reversal of HPR by EHNA was not mediated by the adenosine receptors because the EHNA effect was not blocked by the adenosine receptor antagonist, 8-p-sulfophenyl-theophylline (67 microM; n = 6). Pretreatment with a cy-clic-3',5'-adenosine monophosphate (cAMP)-dependent protein kinase inhibitor, Rp-adenosine-3',5'-Cyclic monophosphorothioate (0.5 mM; n = 4), blocked EHNA reversal of the HPR. As an alternative mechanism of action, EHNA inhibition of Cyclic Nucleotide Phosphodiesterase(s) isozymes was studied in endothelium intact and denuded pulmonary arteries. Using anion-exchange chromatography the Cyclic Nucleotide Phosphodiesterase (PDE) separated into predominantly PDE families 2 and a mixture of 3 and 4. DEAE fractions showing cAMP hydrolysis activated by 5 microM Cyclic-3',5'-guanosine monophosphate (cGMP) had a Km for cAMP of 6.3 microM and an apparent Kact for cGMP of 1.4 microM. EHNA was shown to inhibit PDE2 competitively. In intact vessels, the IC50 for EHNA was 3.3 microM using 0.03 microM [3H]-cAMP substrate assayed in the presence of 2 microM cGMP and in denuded vessels 3.7 microM at 0.03 microM [3H]-cAMP substrate in the presence of 5 microM cGMP. Fractions in which cAMP hydrolysis was inhibited or not affected by 5 microM cGMP (PDE3 and 4, respectively) showed an IC50 of > 200 microM for EHNA. We conclude that reversal of the hypoxic pressor response by EHNA in the isolated, perfused rat lung model occurs with a mechanism involving in part inhibition of smooth muscle PDE2.
P D Peterson - One of the best experts on this subject based on the ideXlab platform.
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erythro 9 2 hydroxy 3 nonyl adenine inhibits Cyclic 3 5 guanosine monophosphate stimulated Phosphodiesterase to reverse hypoxic pulmonary vasoconstriction in the perfused rat lung
Journal of Pharmacology and Experimental Therapeutics, 1996Co-Authors: J Haynes, D W Killilea, P D Peterson, W J ThompsonAbstract:Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was shown to reverse the hypoxic pressor response (HPR) in the isolated, blood-perfused rat lung model. EHNA, an adenosine deaminase inhibitor, showed reversal of the HPR in a dose-dependent manner (EC50 = 129 +/- 30 microM). We found that the reversal of HPR by EHNA was not mediated by the adenosine receptors because the EHNA effect was not blocked by the adenosine receptor antagonist, 8-p-sulfophenyl-theophylline (67 microM; n = 6). Pretreatment with a cy-clic-39,59-adenosine monophosphate (cAMP)-dependent protein kinase inhibitor, Rp-adenosine-39,59-Cyclic monophosphorothioate (0.5 mM; n = 4), blocked EHNA reversal of the HPR. As an alternative mechanism of action, EHNA inhibition of Cyclic Nucleotide Phosphodiesterase(s) isozymes was studied in endothelium intact and denuded pulmonary arteries. Using anion-exchange chromatography the Cyclic Nucleotide Phosphodiesterase (PDE) separated into predominantly PDE families 2 and a mixture of 3 and 4. DEAE fractions showing cAMP hydrolysis activated by 5 microM Cyclic-39,59-guanosine monophosphate (cGMP) had a Km for cAMP of 6.3 microM and an apparent Kact for cGMP of 1.4 microM. EHNA was shown to inhibit PDE2 competitively. In intact vessels, the IC50 for EHNA was 3.3 microM using 0.03 microM [3H]-cAMP substrate assayed in the presence of 2 microM cGMP and in denuded vessels 3.7 microM at 0.03 microM [3H]-cAMP substrate in the presence of 5 microM cGMP. Fractions in which cAMP hydrolysis was inhibited or not affected by 5 microM cGMP (PDE3 and 4, respectively) showed an IC50 of > 200 microM for EHNA. We conclude that reversal of the hypoxic pressor response by EHNA in the isolated, perfused rat lung model occurs with a mechanism involving in part inhibition of smooth muscle PDE2.
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erythro 9 2 hydroxy 3 nonyl adenine inhibits Cyclic 3 5 guanosine monophosphate stimulated Phosphodiesterase to reverse hypoxic pulmonary vasoconstriction in the perfused rat lung
Journal of Pharmacology and Experimental Therapeutics, 1996Co-Authors: J Haynes, D W Killilea, P D Peterson, W J ThompsonAbstract:Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was shown to reverse the hypoxic pressor response (HPR) in the isolated, blood-perfused rat lung model. EHNA, an adenosine deaminase inhibitor, showed reversal of the HPR in a dose-dependent manner (EC50 = 129 +/- 30 microM). We found that the reversal of HPR by EHNA was not mediated by the adenosine receptors because the EHNA effect was not blocked by the adenosine receptor antagonist, 8-p-sulfophenyl-theophylline (67 microM; n = 6). Pretreatment with a cy-clic-3',5'-adenosine monophosphate (cAMP)-dependent protein kinase inhibitor, Rp-adenosine-3',5'-Cyclic monophosphorothioate (0.5 mM; n = 4), blocked EHNA reversal of the HPR. As an alternative mechanism of action, EHNA inhibition of Cyclic Nucleotide Phosphodiesterase(s) isozymes was studied in endothelium intact and denuded pulmonary arteries. Using anion-exchange chromatography the Cyclic Nucleotide Phosphodiesterase (PDE) separated into predominantly PDE families 2 and a mixture of 3 and 4. DEAE fractions showing cAMP hydrolysis activated by 5 microM Cyclic-3',5'-guanosine monophosphate (cGMP) had a Km for cAMP of 6.3 microM and an apparent Kact for cGMP of 1.4 microM. EHNA was shown to inhibit PDE2 competitively. In intact vessels, the IC50 for EHNA was 3.3 microM using 0.03 microM [3H]-cAMP substrate assayed in the presence of 2 microM cGMP and in denuded vessels 3.7 microM at 0.03 microM [3H]-cAMP substrate in the presence of 5 microM cGMP. Fractions in which cAMP hydrolysis was inhibited or not affected by 5 microM cGMP (PDE3 and 4, respectively) showed an IC50 of > 200 microM for EHNA. We conclude that reversal of the hypoxic pressor response by EHNA in the isolated, perfused rat lung model occurs with a mechanism involving in part inhibition of smooth muscle PDE2.
D W Killilea - One of the best experts on this subject based on the ideXlab platform.
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erythro 9 2 hydroxy 3 nonyl adenine inhibits Cyclic 3 5 guanosine monophosphate stimulated Phosphodiesterase to reverse hypoxic pulmonary vasoconstriction in the perfused rat lung
Journal of Pharmacology and Experimental Therapeutics, 1996Co-Authors: J Haynes, D W Killilea, P D Peterson, W J ThompsonAbstract:Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was shown to reverse the hypoxic pressor response (HPR) in the isolated, blood-perfused rat lung model. EHNA, an adenosine deaminase inhibitor, showed reversal of the HPR in a dose-dependent manner (EC50 = 129 +/- 30 microM). We found that the reversal of HPR by EHNA was not mediated by the adenosine receptors because the EHNA effect was not blocked by the adenosine receptor antagonist, 8-p-sulfophenyl-theophylline (67 microM; n = 6). Pretreatment with a cy-clic-39,59-adenosine monophosphate (cAMP)-dependent protein kinase inhibitor, Rp-adenosine-39,59-Cyclic monophosphorothioate (0.5 mM; n = 4), blocked EHNA reversal of the HPR. As an alternative mechanism of action, EHNA inhibition of Cyclic Nucleotide Phosphodiesterase(s) isozymes was studied in endothelium intact and denuded pulmonary arteries. Using anion-exchange chromatography the Cyclic Nucleotide Phosphodiesterase (PDE) separated into predominantly PDE families 2 and a mixture of 3 and 4. DEAE fractions showing cAMP hydrolysis activated by 5 microM Cyclic-39,59-guanosine monophosphate (cGMP) had a Km for cAMP of 6.3 microM and an apparent Kact for cGMP of 1.4 microM. EHNA was shown to inhibit PDE2 competitively. In intact vessels, the IC50 for EHNA was 3.3 microM using 0.03 microM [3H]-cAMP substrate assayed in the presence of 2 microM cGMP and in denuded vessels 3.7 microM at 0.03 microM [3H]-cAMP substrate in the presence of 5 microM cGMP. Fractions in which cAMP hydrolysis was inhibited or not affected by 5 microM cGMP (PDE3 and 4, respectively) showed an IC50 of > 200 microM for EHNA. We conclude that reversal of the hypoxic pressor response by EHNA in the isolated, perfused rat lung model occurs with a mechanism involving in part inhibition of smooth muscle PDE2.
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erythro 9 2 hydroxy 3 nonyl adenine inhibits Cyclic 3 5 guanosine monophosphate stimulated Phosphodiesterase to reverse hypoxic pulmonary vasoconstriction in the perfused rat lung
Journal of Pharmacology and Experimental Therapeutics, 1996Co-Authors: J Haynes, D W Killilea, P D Peterson, W J ThompsonAbstract:Erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) was shown to reverse the hypoxic pressor response (HPR) in the isolated, blood-perfused rat lung model. EHNA, an adenosine deaminase inhibitor, showed reversal of the HPR in a dose-dependent manner (EC50 = 129 +/- 30 microM). We found that the reversal of HPR by EHNA was not mediated by the adenosine receptors because the EHNA effect was not blocked by the adenosine receptor antagonist, 8-p-sulfophenyl-theophylline (67 microM; n = 6). Pretreatment with a cy-clic-3',5'-adenosine monophosphate (cAMP)-dependent protein kinase inhibitor, Rp-adenosine-3',5'-Cyclic monophosphorothioate (0.5 mM; n = 4), blocked EHNA reversal of the HPR. As an alternative mechanism of action, EHNA inhibition of Cyclic Nucleotide Phosphodiesterase(s) isozymes was studied in endothelium intact and denuded pulmonary arteries. Using anion-exchange chromatography the Cyclic Nucleotide Phosphodiesterase (PDE) separated into predominantly PDE families 2 and a mixture of 3 and 4. DEAE fractions showing cAMP hydrolysis activated by 5 microM Cyclic-3',5'-guanosine monophosphate (cGMP) had a Km for cAMP of 6.3 microM and an apparent Kact for cGMP of 1.4 microM. EHNA was shown to inhibit PDE2 competitively. In intact vessels, the IC50 for EHNA was 3.3 microM using 0.03 microM [3H]-cAMP substrate assayed in the presence of 2 microM cGMP and in denuded vessels 3.7 microM at 0.03 microM [3H]-cAMP substrate in the presence of 5 microM cGMP. Fractions in which cAMP hydrolysis was inhibited or not affected by 5 microM cGMP (PDE3 and 4, respectively) showed an IC50 of > 200 microM for EHNA. We conclude that reversal of the hypoxic pressor response by EHNA in the isolated, perfused rat lung model occurs with a mechanism involving in part inhibition of smooth muscle PDE2.
