Effects of SCA40 on human isolated bronchus and human polymorphonuclear leukocytes: comparison with rolipram, SKF94120 and levcromakalim

Cortijo, Julio; Villagrasa, Victoria; Navarrete, C.; Sanz, Celia; Berto, Luisa; Michel, Alain; Bonnet, Pierre-Antoine; Morcillo, E

doi:10.1111/j.1476-5381.1996.tb15682.x

Cited by 35 publications

(31 citation statements)

References 38 publications

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“…PDE activity was measured as described by Schudt et al [16] with modifications [25]. A suspension of 10 7 cells per ml of buffer A (composition in mM: Bis Tris 20, sodium acetate 50, benzamidine 2, EDTA 2, b-mercaptoethanol 5, and phenylmethylsulphonylfluoride (PMSF) 0.05; pH 6.5) was homogenized by sonication (6 ¥ 10 pulses), centrifuged (15 000 g, 20 min), and the supernatant was applied to a Mono-Q HR 5/5 column (1 ml of gel bed, Pharmacia) attached to an FPLC chromatography system.…”

Section: Measurement Of Pde Activity and Camp Contentmentioning

confidence: 99%

Histamine up-regulates phosphodiesterase 4 activity and reduces prostaglandin E 2 -inhibitory effects in human neutrophils

Dası́

Ortiz

Cortijo

et al. 2000

Inflammation Research

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Section: Measurement Of Pde Activity and Camp Contentmentioning

confidence: 99%

Histamine up-regulates phosphodiesterase 4 activity and reduces prostaglandin E 2 -inhibitory effects in human neutrophils

Dası́

Ortiz

Cortijo

et al. 2000

Inflammation Research

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“…Then, either vehicle or fMLP (30 n M ; approx. EC 50 according to previous experiments [20]) was added. The initial rate of signal increase was determined and the enzyme rate was calculated using methylcoumarin amide as a standard.…”

Section: Methodsmentioning

confidence: 99%

Neutrophil Activation in Severe, Early-Onset COPD Patients versus Healthy Non-Smoker Subjects in vitro: Effects of Antioxidant Therapy

Milara¹,

Juan²,

Peiró³

et al. 2011

Respiration

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Background: Neutrophils and oxidative stress have been implicated in the pathogenesis of COPD. Severe, early-onset COPD is characterized by a rapid decline in the lung function at an early age; however, nothing is known about neutrophil activation in COPD patients. Objectives: The aim of this study was to evaluate peripheral blood neutrophil activation in severe, early-onset COPD patients versus healthy non-smokers and the effect of N-acetyl-L-cysteine (NAC) on neutrophil activation in vitro. Methods: Neutrophils were isolated from 15 severe, early-onset COPD patients and 15 age-matched healthy subjects and stimulated with N-formyl- Met-Leu-Phe (fMLP) in the presence or absence of NAC (10 µM to 10 mM). Neutrophil chemotaxis, elastase release, reactive oxygen species (ROS), intracellular thiols and apoptosis were measured by Boyden chamber, spectrofluorometry, CMFDA and H₂DCF-DA dyes and by annexin V-FITC, respectively. Results: Chemotaxis of peripheral blood neutrophils from COPD patients in response to fMLP was 30% more increased than that observed in healthy subjects. Elastase release in response to fMLP was 2-fold higher in neutrophils from COPD patients versus healthy subjects. Intracellular thiol levels were 30% lower in COPD and ROS was approximately 30% higher in COPD versus healthy neutrophils. Spontaneous apoptosis showed no differences in both groups of patients and fMLP-induced apoptosis was higher in COPD. Pre-treatment with the antioxidant NAC effectively attenuated neutrophil chemotaxis, elastase release and ROS as well as effectively increased thiol levels in COPD. Conclusions: Neutrophils in severe, early-onset COPD patients are highly activated and this is alleviated by NAC in vitro.

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“…For the same level of stimulated tone, the loss of the inhibitory potency of SCA40 was more pronounced in those preparations contracted with acetylcholine. SCA40 is a potent inhibitor of PDE 3 while its potency as inhibitor of PDE 4 and PDE 5 was one to two orders of magnitude less (Cook et al 1995;Cortijo et al 1996). The range of potencies of SCA40 as inhibitor of spontaneous and stimulated bronchial tone (-log IC 50~ 6.5 to 5) was within its range of potencies as inhibitor of PDE 3 and 4 activities (-log IC 50~ 6.5 to 5; Cortijo et al 1996).…”

Section: Relaxation Of Human Isolated Bronchus By Sca40mentioning

confidence: 98%

“…SCA40 is a potent inhibitor of PDE 3 while its potency as inhibitor of PDE 4 and PDE 5 was one to two orders of magnitude less (Cook et al 1995;Cortijo et al 1996). The range of potencies of SCA40 as inhibitor of spontaneous and stimulated bronchial tone (-log IC 50~ 6.5 to 5) was within its range of potencies as inhibitor of PDE 3 and 4 activities (-log IC 50~ 6.5 to 5; Cortijo et al 1996). A number of selective PDE 3 and PDE 4 inhibitors have been consistently reported to inhibit the spontaneous and stimulated tone of human bronchial preparations with -log IC 50 values in the range of those reported for inhibition of their corresponding PDE activities (De Boer et al 1992;Torphy et al 1993;Cortijo et al 1993Cortijo et al , 1996Naline et al 1996b).…”

Section: Relaxation Of Human Isolated Bronchus By Sca40mentioning

confidence: 99%

“…Subsequently, Macmillan et al (1995) demonstrated in patch clamp studies that SCA40 (1-10 µM) was not opening BK Ca , and Pocock and Small (1996) showed that SCA40 failed to stimulate 86 Rb + efflux from bovine trachea. Cook et al (1995) and Cortijo et al (1996) showed that SCA40 is a potent inhibitor of PDE type 3 isoenzyme .5) but it was also inhibiting PDE 4 (-log IC 50~ 4.8) and PDE 5 (-log IC 50~ 4.3) isoenzymes. Therefore, an elevation in the cellular content of cyclic nucleotides may be underlying relaxation of guinea-pig and bovine trachea Pocock and Small 1996) as well as of human isolated bronchus Naline et al 1996a).…”

Section: Introductionmentioning

confidence: 99%

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Bronchodilator and anti-inflammatory activities of SCA40: Studies in human isolated bronchus, human eosinophils, and in the guinea-pig in vivo

Cortijo

Pons

Dası́

et al. 1997

Naunyn-Schmiedeberg's Arch Pharmacol

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There is currently interest in the use of inhibitors of cyclic nucleotide phosphodiesterases (PDE) as potential anti-asthma agents. In this study we examined the effects of SCA40 (6-bromo-8-methylaminoimidazol-[1,2-a] pyrazine-2-carbonitrile), a preferential inhibitor of PDE 3 also endowed with PDE 4 and 5 inhibitory activities, on isolated bronchus and eosinophil functions and in an animal model of asthma. SCA40 (1 nM-0.1 mM) produced concentration-dependent inhibition of spontaneous and stimulated tone of human isolated bronchus and reached a maximal relaxation similar to that of theophylline (3 mM). The potency (-log EC50 values) of SCA40 against spontaneous tone (6.52 +/- 0.10) was greater than against tone raised by equieffective concentrations (approximately 70%) of histamine (5.76 +/- 0.06), leukotriene C4 (5.44 +/- 0.11), and acetylcholine (4.98 +/- 0.09). In the presence of cytochalasin B, the chemotactic peptide N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP; 0.5 microM) induced leukotriene C4 production in human eosinophils isolated in discontinuous metrizamide gradients. The production of leukotriene C4 was inhibited by SCA40 in a concentration-related fashion (-log IC50 = 6.04 +/- 0.20; n = 6). Rolipram, a selective PDE 4 inhibitor, was also effective (-log IC50 = 7.29 +/- 0.32) but the selective PDE 3 inhibitor SKF94120 was scarcely effective (< 10% inhibition for 10 microM). In ovalbumin sensitized guinea-pigs, SCA40 (1 mg kg(-1), i.p.) given 30 min before antigen challenge significantly inhibited the acute bronchoconstriction produced by aerosol antigen (5 mg ml(-1), 30 s) (antigen response was 185 +/- 13 and 91 +/- 21 cmH2O l(-1) s(-1) in control and SCA40-treated animals, respectively, P < 0.05). Pretreatment with SCA40 (1 mg kg(-1), i.p., 30 min pre- and 3 h post-antigen exposure) prevented airway hyperreactivity to histamine which developed 24 h after exposure of conscious guinea-pigs to aerosol antigen. Eosinophil lung accumulation that accompanied airway hyperreactivity was also inhibited by SCA40 (from 6.15 +/- 0.86 in control to 1.27 +/- 0.27 in treated animals; expressed as eosinophils x 10(6); P < 0.05). SCA40 (1 mg kg(-1), i.p.) also inhibited the microvascular leakage produced after inhaled antigen (5 mg ml(-1), 30 s) at all airway levels. The haemodynamic effects of SCA40 (1 mg kg(-1), i.p.) consisted of a rapid decrease (peak at 5 min) in mean arterial blood pressure (-39.4 +/- 2.4%) and tracheal mucosal blood flow (-13.5 +/- 2.0%) that slowly recovered with time. These data support previous work showing that PDE inhibition results in antispasmogenic and anti-inflammatory effects. SCA40 was effective in vitro and in vivo and these effects are probably related to its activity as a mixed PDE inhibitor.

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Effects of SCA40 on human isolated bronchus and human polymorphonuclear leukocytes: comparison with rolipram, SKF94120 and levcromakalim

Cited by 35 publications

References 38 publications

Histamine up-regulates phosphodiesterase 4 activity and reduces prostaglandin E 2 -inhibitory effects in human neutrophils

Histamine up-regulates phosphodiesterase 4 activity and reduces prostaglandin E 2 -inhibitory effects in human neutrophils

Neutrophil Activation in Severe, Early-Onset COPD Patients versus Healthy Non-Smoker Subjects in vitro: Effects of Antioxidant Therapy

Bronchodilator and anti-inflammatory activities of SCA40: Studies in human isolated bronchus, human eosinophils, and in the guinea-pig in vivo

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