Guinea Pig Trachea Research Articles

Interaction of prostanoid EP3 and TP receptors in guinea-pig isolated aorta: contractile self-synergism of 11-deoxy-16,16-dimethyl PGE2

Surprisingly high contractile activity was reported for 11-deoxy-16,16-dimethyl prostaglandin E₂ (DX-DM PGE₂) on pig cerebral artery when used as a selective EP₃ receptor agonist. This study investigated the selectivity profile of DX-DM PGE₂, focusing on the interaction between its EP₃ and TP (thromboxane A₂-like) agonist activities. Contraction of guinea-pig trachea (EP₁ system) and aorta (EP₃ and TP systems) was measured in conventional organ baths. Strong contraction of guinea-pig aorta to sulprostone and 17-phenyl PGE₂ (EP₃ agonists) was only seen under priming with a second contractile agent such as phenylephrine, histamine or U-46619 (TP agonist). In contrast, DX-DM PGE₂ induced strong contraction, which on the basis of treatment with (DG)-3ap (EP₃ antagonist) and/or BMS-180291 (TP antagonist) was attributed to self-synergism arising from co-activation of EP₃ and TP receptors. EP₃/TP self-synergism also accounted for contraction induced by PGF(2α) and its analogues (+)-cloprostenol and latanoprost-FA. DX-DM PGE₂ also showed significant EP₁ agonism on guinea-pig trachea as defined by the EP₁ antagonists SC-51322, (ONO)-5-methyl-1 and AH-6809, although AH-6809 exhibited poor specificity at concentrations ≥3 µM. EP₃/TP self-synergism, as seen with PGE/PGF analogues in this study, may confound EP₃ agonist potency comparisons and the characterization of prostanoid receptor systems. The competitive profile of a TP antagonist may be distorted by variation in the silent/overt contraction profile of the EP₃ system in different studies. The relevance of self-synergism to in vivo actions of natural prostanoid receptor agonists is discussed.

Zataria multiflora Boiss and Carvacrol Affect β(2)-Adrenoceptors of Guinea Pig Trachea.

The stimulatory effect of Zataria multiflora Boiss (Labiatae) and carvacrol on β-adrenoceptors was examined on guinea pig trachea. The effects of three concentrations of aqueous-ethanolic extract, carvacrol, and propranolol (β-receptor antagonist) on β-adrenoceptors were tested in nonincubated (group 1, n = 8) and incubated tracheal chains with 1 μM chlorpheniramine (histamine H1 receptor antagonist) (group 2, n = 5). Isoprenaline (β-receptor agonist) curves obtained in the presence of all concentrations of the extract and carvacrol showed leftward shifts compared with that of saline in both groups. In both groups, the EC50 (the effective concentration of isoprenaline, causing 50% of maximum response) obtained in the presence of all concentrations of the extract and carvacrol was significantly lower compared to that of saline (P < .01 to P < .001). All values of (CR-1: (EC50 in the presence of active substances/EC50 obtained in the presence of saline)-1) obtained in the presence of concentrations of the extract and carvacrol in both groups were negative and significantly different from that of propranolol (P < .001 for all cases). The results indicated a stimulatory effect of Zataria multiflora Boiss extract on β 2-adrenoceptors which is perhaps due to its constituent, carvacrol.

Expression and function of human hemokinin-1 in human and guinea pig airways

BackgroundHuman hemokinin-1 (hHK-1) and endokinins are peptides of the tachykinin family encoded by the TAC4 gene. TAC4 and hHK-1 expression as well as effects of hHK-1 in the lung and airways remain however unknown and were explored in this study.MethodsRT-PCR analysis was performed on human bronchi to assess expression of tachykinin and tachykinin receptors genes. Enzyme immunoassay was used to quantify hHK-1, and effects of hHK-1 and endokinins on contraction of human and guinea pig airways were then evaluated, as well as the role of hHK-1 on cytokines production by human lung parenchyma or bronchi explants and by lung macrophages.ResultsIn human bronchi, expression of the genes that encode for hHK-1, tachykinin NK1-and NK2-receptors was demonstrated. hHK-1 protein was found in supernatants from explants of human bronchi, lung parenchyma and lung macrophages. Exogenous hHK-1 caused a contractile response in human bronchi mainly through the activation of NK2-receptors, which blockade unmasked a NK1-receptor involvement, subject to a rapid desensitization. In the guinea pig trachea, hHK-1 caused a concentration-dependant contraction mainly mediated through the activation of NK1-receptors. Endokinin A/B exerted similar effects to hHK-1 on both human bronchi and guinea pig trachea, whereas endokinins C and D were inactive. hHK-1 had no impact on the production of cytokines by explants of human bronchi or lung parenchyma, or by human lung macrophages.ConclusionsWe demonstrate endogenous expression of TAC4 in human bronchi, the encoded peptide hHK-1 being expressed and involved in contraction of human and guinea pig airways.

Effect of the Aqueous Extract of &lt;i&gt;Crinum glaucum&lt;/i&gt; on Isolated Guinea Pig Trachea

The aqueous extract of Crinum glaucum was evaluated using guinea pig trachea and was observed to neither relax the spontaneous tone of the guinea pig trachea nor KCl-induced tone. It, however, produced a concentration-dependent relaxation of carbachol- and histamine- pre-contracted trachea. The relaxant effect of the extract on carbachol-induced tone was not inhibited by propranolol, but potentiated by glibenclamide, procaine, methylene blue and L-nitro argininemethyl ester (L-NAME) with a progressive decrease in the median effective concentration (EC50) values. The extract (5-40 mg/kg body weight/day) produced a rightward shift and a depression of the maximal effects (Emax) of the anaphylactic contractions of the trachea while the extract at 2mg/ml produced a complete inhibition of the contractions.The results of this study clearly showed that the aqueous extract of Crinum glaucum possesses spasmolytic and anti-spasmogenic properties on normal and sensitized trachea and this may account for its use in the treatment of cough, asthma, and convulsions in traditional medicine.Key words: Crinum glaucum, trachea, anaphylactic contractions, histamine, carbachol

Calcium channel subtypes for cholinergic and nonadrenergic noncholinergic neurotransmission in isolated guinea pig trachea

The Ca(2+) channel subtypes in the neurotransmission of isolated guinea pig trachea were elucidated by monitoring the effects of specific Ca(2+) channel blockers on cholinergic contractions and nonadrenergic noncholinergic (NANC) relaxation elicited by electrical field stimulation (EFS). In isolated guinea pig trachea, cholinergic contractile responses to low- and high-frequency EFS were inhibited by the selective N-type calcium channel blocker, ω-conotoxin MVIIA. ω-Agatoxin IVA (a selective P-type blocker), ω-conotoxin MVIIC (a nonselective N-, Q-, and P-type blocker), and nifedipine (a selective L-type blocker) were ineffective, whereas Ni(2+) (a T- and R-type blocker) facilitated cholinergic contractions and produced a late contracture when its concentration exceeded 30μM. The more the concentration of Ni(2+) increased, the greater the number of incidences and the late contracture areas which occurred. Late contracture did not seem to be due to the effects of acetylcholine, tachykinins, or other polypeptides, but disappeared in the absence of indomethacin. The NANC relaxant responses elicited by the low- and high-frequency EFS were inhibited by ω-conotoxin MVIIA or Ni(2+), but unaffected by ω-Agatoxin IVA, ω-conotoxin MVIIC, and nifedipine. In the absence of indomethacin, Ni(2+) did not alter the ω-conotoxin MVIIA (100nM)-resistant component of cholinergic contraction, but significantly further inhibited that of NANC relaxation. These results suggest that in isolated guinea pig trachea, cholinergic contraction is regulated by N-type calcium channels which may mask T- and R-type calcium channels and may be co-modulated by both, while NANC relaxation is mainly and independently controlled by N-, T-, and R-type calcium channels.

Pharmacological studies on Hypericum perforatum fractions and constituents

Context: This study describes the antispasmodic, bronchodilator, and cardiovascular-modulatory activities of Hypericum perforatum Linn. (Hypericaceae) fractions and constituents.Aim of study: Pharmacological investigation of H. perforatum fractions and active principles.Materials and methods: H. perforatum extract fractions [petroleum spirit (HpPet), chloroform (HpCHCl3), ethyl acetate (HpEtAc), and aqueous (HpAq)] and its compounds (hyperforin, hypericin, and hyperoside) were studied in various isolated tissue preparations.Results: In rabbit jejunum, HpCHCl3, HpEtAc and HpAq, like papaverine, inhibited both spontaneous and K+ (80 mM)-induced contractions at similar concentrations, whereas HpPet was relatively potent against K+, as verapamil. All fractions caused rightward of Ca2+ concentration-response curves (CRCs), similar to verapamil. HpCHCl3, HpEtAc, and HpAq shifted isoprenaline-inhibitory CRCs to left, like papaverine, while HpPet was devoid of any such effect, as verapamil. In guinea-pig trachea, HpCHCl3, HpEtAc, and HpAq equipotently relaxed carbachol and K+-induced contractions and shifted the isoprenaline-curves to the left, whereas HpPet was more effective against K+, without potentiating isoprenaline effect. When tested in rabbit aorta, all fractions exhibited vasoconstrictor and vasodilator effects, except HpEtAc, which did not produce vasoconstriction. In guinea-pig atria HpCHCl3, HpEtAc, and HpAq initially caused cardiac stimulation, followed by inhibition, similar to papaverine, whereas HpPet, like verapamil, caused only cardiac suppression. Hyperforin, hypericin, and hyperoside showed a similar pattern of spasmolytic effect to verapamil.Discussion and conclusion: Thus, all tested fractions of H. perforatum exhibit a combination of Ca2+ antagonist and phosphodiesterase-inhibition, except petroleum spirit which was devoid of later mechanism. The compounds tested showed only Ca2+ channel blocking effect.

Bronchodilatory effect of Acorus calamus (Linn.) is mediated through multiple pathways

Aim of the study This study was undertaken to provide a pharmacological basis for traditional use of Acorus calamus in airways disorders. Materials and methods Isolated guinea-pig trachea and atria were suspended in organ baths bubbled with carbogen and mechanisms were found using different parameters. Results In isolated guinea-pig tracheal segments, crude extract of Acorus calamus was more effective than carbachol in causing relaxation of high K + (80 mM) precontractions, similar to verapamil, suggesting blockade of calcium channels. The n-hexane fraction was equipotent against both precontractions, similar to papaverine, while ethylacetate fraction was more potent against carbachol precontractions but had a negligible dilator effect against K +, similar to atropine and or rolipram. Pretreatment of tracheal preparations with n-hexane or ethylacetate fractions potentiated isoprenaline-induced inhibitory concentration–response curves, similar to papaverine or rolipram. Pretreatment of tracheal preparations with ethylacetate fraction caused a rightward parallel shift in carbachol response curve at lower concentration (0.003 mg/mL) similar to atropine and a non-parallel shift at higher concentrations (0.01 mg/mL), with reduction of maximum response, similar to rolipram. In isolated guinea-pig atrial preparations, crude extracts, its fractions and papaverine inhibited force and rate of contractions at higher concentrations than the smooth muscle while verapamil was equipotent. Conclusion These data indicate the presence of unique combination of airways relaxant constituents in crude extract of Acorus calamus, a papaverine-like dual inhibitor of calcium channels and phosphodiesterase in n-hexane fraction and a novel combination of anticholinergic, rolipram-like phosphodiesterase4 inhibitor in ethylacetate fraction and associated cardiac depressant effect, provide a pharmacological basis for traditional use of Acorus calamus in disorders of airways.

Propofol Preferentially Relaxes Neurokinin Receptor-2-induced Airway Smooth Muscle Contraction in Guinea Pig Trachea

Propofol is the anesthetic of choice for patients with reactive airway disease and is thought to reduce intubation- or irritant-induced bronchoconstriction by decreasing the cholinergic component of vagal nerve activation. However, additional neurotransmitters, including neurokinins, play a role in irritant-induced bronchoconstriction. We questioned the mechanistic assumption that the clinically recognized protective effect of propofol against irritant-induced bronchoconstriction during intubation was due to attenuation of airway cholinergic reflexes. Muscle force was continuously recorded from isolated guinea pig tracheal rings in organ baths. Rings were subjected to exogenous contractile agonists (acetylcholine, histamine, endothelin-1, substance P, acetyl-substance P, and neurokinin A) or to electrical field stimulation (EFS) to differentiate cholinergic or nonadrenergic, noncholinergic nerve-mediated contraction with or without cumulatively increasing concentrations of propofol, thiopental, etomidate, or ketamine. Propofol did not attenuate the cholinergic component of EFS-induced contraction at clinically relevant concentrations. In contrast, propofol relaxed nonadrenergic, noncholinergic-mediated EFS contraction at concentrations within the clinical range (20-100 mum, n = 9; P < 0.05), and propofol was more potent against an exogenous selective neurokinin-2 receptor versus neurokinin-1 receptor agonist contraction (n = 6, P < 0.001). Propofol, at clinically relevant concentrations, relaxes airway smooth muscle contracted by nonadrenergic, noncholinergic-mediated EFS and exogenous neurokinins but not contractions elicited by the cholinergic component of EFS. These findings suggest that the mechanism of protective effects of propofol against irritant-induced bronchoconstriction involves attenuation of tachykinins released from nonadrenergic, noncholinergic nerves acting at neurokinin-2 receptors on airway smooth muscle.

Synthesis and Structure−Activity Relationships of Long-acting β2 Adrenergic Receptor Agonists Incorporating Metabolic Inactivation: An Antedrug Approach

A series of saligenin beta(2) adrenoceptor agonist antedrugs having high clearance were prepared by reacting a protected saligenin oxazolidinone with protected hydroxyethoxyalkoxyalkyl bromides, followed by removal of the hydroxy-protecting group, alkylation, and final deprotection. The compounds were screened for beta(2), beta(1), and beta(3) agonist activity in CHO cells. The onset and duration of action in vitro of selected compounds were assessed on isolated superfused guinea pig trachea. Compound 13f had high potency, selectivity, fast onset, and long duration of action in vitro and was found to have long duration in vivo, low oral bioavailability in the rat, and to be rapidly metabolized. Crystalline salts of 13f (vilanterol) were identified that had suitable properties for inhaled administration. A proposed binding mode for 13f to the beta(2)-receptor is presented.

Antioxidant, Anti-Proliferative And Bronchodilatory Activities Of Euphorbia Hirta Extracts

Euphorbia hirta is a plant commonly found in wild lands in tropical countries. It has various medicinal uses such as for conjunctivitis, ulcerated cornea, asthma, bronchitis, upper respiratory catarrh etc.&nbsp; In this study, crude n-hexane, chloroform, methanol and water extracts of the plant were screened for their antioxidant activities using an improved 2, 2’-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical cation decolorisation assay method.&nbsp; The methanol and water extracts showed comparable antioxidant activities to those of green and black teas.&nbsp; On the other hand, the chloroform and n-hexane extracts did not show significant antioxidant activities.&nbsp; The extracts were also screened for anti-proliferative acitivities against normal mouse fibroblast cells (NIH/3T3).&nbsp; Results indicated that the methanol and chloroform extracts were less anti-proliferative than the water and hexane extracts.&nbsp; The effect of Euphorbia hirta aqueous extract on guinea pig trachea was studied.&nbsp; The water extract alone did not have any effect on the guinea pig trachea but it showed partial inhibition on the histamine induced tracheal smooth muscles contractions.&nbsp; Therefore, further studies may focus on water extract as it showed considerable antioxidant activity and partial inhibitory effect on histamine induced tracheal smooth muscle contractions.&nbsp; &nbsp; (Keywords: &nbsp;Antioxidant activity; Bronchodilation; Anti-proliferative activity; Euphorbia hirta; Plant extracts.) &nbsp;

Gnaphaliin A and Gnaphaliin B Identified as Active Principles of Gnaphalium liebmannii Sch. B.p. ex Klatt (Asteraceae) with Tracheal Smooth Muscle Relaxant Properties

Inflorescences of Gnaphalium liebmannii Sch. B.p. ex Klatt, commonly known as „Gordolobo“, is the most important remedy in Mexican traditional medicine to treat respiratory diseases, including asthma. Using a bio-guided fractionation of the n-hexane extract of this plant, following the relaxant effect on guinea pig tracheal smooth muscle, the flavones 5,7-dihydroxy-3,8-dimethoxyflavone (1) and 3,5-dihydroxy-7,8-dimethoxyflavone (2) were identified as the active relaxant compounds. Both 1 and 2 have been described as gnaphaliin in the past; The EIMS data, NMR experiments for both compounds, and X-ray diffraction analysis for 1, provided structural information to suggest that 1 and 2 should be named gnaphaliins A and B, respectively. Gnaphaliin A and gnaphaliin B induced relaxation on guinea pig trachea with EC50 values of 181.58±2.22µM and 128.36±1.88µM respectively. Both flavones were more potent to relax the guinea pig trachea than enoximone, rolipram and aminophylline, but they were less potent than sildenafil. Gnaphaliin A and B were also effective to inhibit cAMP (IC50=120.8±26µM and 72.1±11µM, respectively) and cGMP (IC50=2.9±0.3µM and 6.4±2µM, respectively) degradation by PDE, showing preference to inhibit the degradation of cGMP, with the follow activity order: sildenafil &gt; rolipram &gt; gnaphaliin A &gt; gnaphaliin B &gt; aminophylline &gt; IBMX, enoximone. In addition, both flavones inhibited the flux of extracellular calcium in guinea-pig trachea. Acknowledgements: This work was supported by grant of Dirección General de Asuntos del Personal Académico DGAPA-UNAM IN 205008 and CONACYT 82613. F. Rodríguez-Ramos acknowledges fellowship from CONACYT 131381.

Effect of an alpha-blocker on the contraction of guinea pig trachea with various bronchoconstrictors (author's transl)

Effect of an alpha-blocker on the contraction of guinea pig trachea with various bronchoconstrictors (author's transl)

Mechanisms involved in the relaxant action of the ethanolic extract of propolis in the guinea-pig trachea in-vitro.

This study examines the mechanisms by which the standardised ethanolic extract of propolis induces relaxation of the guinea-pig trachea in-vitro. In guinea-pig trachea with or without epithelium and contracted by histamine, the propolis extract caused reproducible and graded relaxation, with a mean EC50 value of 3.8 or 10.5 microg mL(-1) and Emax of 100%, respectively. The propolis extract-induced relaxation was markedly reduced (26+/-9 and 96+/-3%) when guinea-pig tracheas were exposed to Krebs solution containing elevated K+ in the medium (40 or 80 mM). Pre-incubation of guinea-pig tracheas with tetraethylamonium (100 mM) or with 4-aminopyridine (10mM) reduced the propolis extract-induced relaxation by 31+/-10% and 28+/-2%. Likewise, apamin (0.1 microM), charybdotoxin (0.1 microM) or iberiotoxin (0.1 microM) caused marked inhibition of propolis extract-mediated relaxation in guinea-pig trachea (percentage of inhibition: 65+/-3%, 60+/-5% and 65+/-9%, respectively). Also, glibenclamide (1 microM) inhibited the relaxant response caused by the propolis extract by 57+/-4%. Omega-conotoxin GIVA (0.1 microM) or capsaicin (1 microM) produced small but significant inhibition (30+/-5% or 47+/-7%, respectively) of the propolis extract-induced relaxation. The vasoactive intestinal peptide (VIP) antagonist D-p-Cl-Phe6,Leu17[VIP] porcine (0.1 microM) inhibited relaxation by 55+/-5%, while propranolol (1 microM) induced a parallel rightward displacement (about 20 fold) of the propolis extract concentration-response curve. Finally, the propolis extract-induced relaxation was inhibited by the nitric oxide synthase inhibitor L-N(G)-nitroarginine (L-NOArg, 100 microM) (48+/-6%), and by the soluble guanylatecyclase inhibitormethylene blue (10 microM) (37+/-6%), whilethe moreselectivesoluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolol[4,3-alquinoxalin-1-one (ODQ, 1 microM) produced only a parallel (about 3 fold) rightward displacement of the propolis extract concentration-response curve. Collectively, these results support the notion that the propolis extract-mediated relaxation in the guinea-pig trachea involves the release of nitric oxide, probably from sensory neurons, besides the activation of soluble guanylate cyclase and activation of Ca2+- and ATP-sensitive K+ channels. Furthermore, the stimulation of beta2-adrenergic and VIP receptors also seems to account for its relaxant action.

Bronchodilator activity of Mikania glomerata Sprengel on human bronchi and guinea-pig trachea.

The effects of aqueous extracts and hydro-alcoholic extract (HAE), and of a dichloromethane fraction (MG1) obtained from the HAE of Mikania glomerata leaves on isolated respiratory and vascular smooth muscle have been investigated. Aqueousextracts and HAE induced a significant inhibition on the histamine contractions on the isolated guinea-pig trachea. HAE extract induced a concentration-dependent relaxation on guinea-pig trachea pre-contracted with histamine (IC50 0.34 (0.29-0.39) mg mL(-1)), acetylcholine (IC50 0.72 (0.67-0.77) mg mL(-1)) or K+ (IC50 1.41 (1.18-1.64) mg mL(-1)) and on isolated human bronchi precontracted with K+ (IC50 0.34 (0.26-0.42) mg mL(-1)). The dichloromethane fraction induced a concentration dependent relaxation in guinea-pig trachea precontracted with K+ (IC50 0.017 (0.012-0.022) mg mL(-1)). The dichloromethane fraction had also a small vasodilator effect on the isolated mesenteric vascular bed and on the isolated rat aorta, and a significant reduction of the oedema induced by subplantar injections of Bothropsjararaca venom in mice. When tested on plasmid DNA, MG1 did not damage the DNA. Chromatographic analysis showed the presence of 11.4% w/w coumarin in MG1. The results supported the indication of M. glomerata products for the treatment of respiratory diseases where bronchoconstriction is present.

β-Blocking activity of PP-34, a newly synthesized aryloxypropanolamine derivative, and its cardioprotective effect against ischaemia/reperfusion injury in laboratory animals

Beta-adrenoceptor antagonists are widely used in cardiovascular medicine. However, the main side effect of these drugs is due to antagonism of beta(2)-adrenoceptors in the airways, resulting in bronchospasm. Therefore, more cardioselective beta-blockers have been developed to offer a lower side effect profile. We have studied a new aryloxypropanolamine derivative (PP-34) with more cardioselectivity and efficacy against ischaemia/reperfusion injury in rats. Oxalate salts of 1-(tert-butylamino)-3-(5-tert-butylaminomethyl-2-methoxyphenoxy) propan-2-ol (PP-34) is a novel beta-adrenoceptor antagonist. In-vitro studies in rat isolated right atria, guinea-pig trachea and rat distal colon preparations were carried out to investigate the potency of PP-34 towards different beta-adrenoceptor subtypes. pA(2)/pK(B) values of PP-34 for beta(1), beta(2), and beta(3) adrenoceptor were 7.89+/-0.15, 6.13+/-0.09 and 6.30+/-0.19, respectively. The beta(1)/beta(2) selectivity ratio calculated was in the order of PP-34 > atenolol > propranolol. Pre-ischaemic administration (20 min before coronary occlusion) of PP-34 (0.3 or 1 mg kg(-1)) showed cardioprotective effects against ischaemia/reperfusion injury in rats and significantly reduced arrhythmias, infarct area and necrosis induced by ischaemia/reperfusion injury. The efficacy of PP-34 was found to be greater then atenolol. In conclusion, PP-34 is a cardioselective beta-adrenoceptor antagonist, possessing potent anti-arrhythmic and cardioprotective effects against ischaemia/reperfusion injury in rats.

Red wine polyphenolic compounds inhibit tracheal smooth muscle contraction during allergen-induced hyperreactivity of the airways

The aims of the study were to investigate the short and long-term effects of Provinol (red wine polyphenolic compounds) on tracheal smooth muscle reactivity using an in-vitro model of ovalbumin-induced airway inflammation in guinea-pig trachea, and to evaluate the role of nitric oxide (NO) in the bronchodilatory effect of Provinol. The amplitude of tracheal smooth muscle contraction in response to mediators of bronchoconstriction - histamine (10 nM-1 mM), acetylcholine (10 nM-1 mM) and to allergen (ovalbumin 10(-5)-10(-3) g mL(-1)) was used as a parameter of tracheal smooth muscle reactivity. To test the short-term effects of Provinol, isolated tracheal strips were pre-treated for 30 min with Provinol (10(-4) mg mL(-1)) alone or in combination with Nomega-nitro-L-arginine methyl ester (L-NAME; 10(-6) mol L(-1)). To test the long-term effects of Provinol, isolated tracheal strips were prepared from guinea pigs that had been treated for 14 days with Provinol (20 mg kg(-1) per day) alone or in combination with L-NAME (40 mg kg(-1) per day). Incubation of tracheal smooth muscle with Provinol decreased the amplitude of contraction in response to ovalbumin, histamine and acetylcholine. The non-selective NO synthase inhibitor L-NAME partially abolished the effect of Provinol on acetylcholine and ovalbumin-induced but not histamine-induced bronchoconstriction. A similar profile was observed after 14 days' oral administration of Provinol. In conclusion, Provinol inhibited the allergen- and spasmogen-induced contraction of tracheal smooth muscle in ovalbumin-sensitized guinea pigs via a mechanism that was mediated at least partially through the metabolism of NO.

Design, synthesis and biological evaluation of a sulfonylcyanoguanidine as thromboxane A2 receptor antagonist and thromboxane synthase inhibitor

The synthesis and the structure of N-isopropyl-N'-[2-(3'-methylphenylamino)-5-nitrobenzenesulfonyl] urea (14) was drawn from two thromboxane A2 receptor antagonists structurally related to torasemide. Compound 14 showed an IC50 value of 22 nM for the thromboxane A2 (TXA2) receptor of human washed platelets. Compound 14 prevented platelet aggregation induced by arachidonic acid (0.6 mM) and U-46619 (1 microM) with an IC50 value of 0.45 and 0.15 microM, respectively. Moreover, 14 relaxed the rat isolated aorta and guinea-pig trachea precontracted by U-46619, a TXA2 agonist. Its efficacy (IC50) was 20.4 and 5.47 nM, respectively. Finally, 14 (1 microM) completely inhibited TXA2 synthase of human platelets. The pKa value and the crystallographic data of 14 were determined and used to propose an interaction model between the TXA2 antagonists related to torasemide and their receptor.

Synthesis and pharmacological evaluation of 5-dialkylaminomethyl-2-amino-2-oxazolines as H1-antagonists.

New 5-dialkylaminomethyl-2-amino-2-oxazolines have been synthezised in two steps from the corresponding dialkylamines. They were evaluated in-vitro as H1-antagonists. Compounds 1c, 1d and 1j significantly antagonized histamine-induced contraction of guinea-pig trachea with a rightward shift of the concentration-response curve to histamine. Compound 1f, 5-[(4-benzyl-1-piperidinyl)methyl]-2-amino-2-oxazoline, induced an increase in acetylcholine Emax (the maximal response to acetylcholine 10(-3) M) and a shift to the left of the concentration-response curve. The lack of effect of this compound on histamine-induced contraction rules out a non-selective potentiation of the contraction mechanisms. Preliminary structure-activity results were reported partly based on physicochemical results.

Cardioprotective and β-adrenoceptor antagonistic activity of a newly synthesized aryloxypropanolamine derivative PP-36.

The present study was performed to evaluate the cardioprotective effects and pharmacological characterization of newly synthesized β-adrenoreceptor antagonists 3-(3-tert-butylamino-2-hydroxypropoxy)-4-methoxybenzaldehyde (PP-36) in the rat model of coronary artery occlusion and reperfusion. Pre-ischemic administration (20 minutes before coronary occlusion) of PP-36 showed cardioprotective effects against ischemia/reperfusion injury in rats. PP-36 (6 mg kg(-1)) significantly reduced arrhythmia score (6.33 ± 0.55, P < 0.05), infarct size/left ventricle size (38.9 ± 3.2, P < 0.05) and no mortality compared to vehicle-treated control group (14.17 ± 1.83, 44.9 ± 4.6 and 17% respectively). In-vitro studies in rat isolated right atria, guinea-pig trachea and rat distal colon preparations, were carried out to investigate the potency of PP-36 towards different β-adrenoceptor subtypes. pA2/pKB values of PP-36 for β1-β2-and β3-adrenoceptors were 6.904 ± 0.190, 6.44 ± 0.129 and 5.773 ± 0.129, respectively. In conclusion, PP-36 is a β-adrenoceptor antagonist possessing potent anti-arrhythmic and cardioprotective effects against ischemia/reperfusion injury in rats.

Pharmacological and phytochemical study on a Sisymbrium officinale Scop. extract

Ethnopharmacological relevance The aerial parts of Sisymbrium officinale Scop. are commonly used to treat airway ailments, moreover in antiquity the herbal drug was reputed to possess anticancer properties. The results obtained in present work support the traditional use and the properties ascribed to Sisymbrium officinale. Aim of the study In order to give a scientific basis to the traditional uses of Sisymbrium officinale, this study was aimed to evaluate in vitro the myorelaxant activity, the antimicrobial properties and the antimutagenic effect of an aqueous dry extract of the aerial parts of the plant. A phytochemical characterization of the extract was also performed. Materials and methods The myorelaxant activity was studied against the contractions induced by carbachol, histamine and leukotriene C 4, in isolated guinea-pig trachea. The antimicrobial activity was tested against six bacteria and one yeast. The Ames test, performed by the preincubation method, was used to study the antimutagenic activity of the extract by its capability to inhibit the mutagenic effect of 2-nitrofluorene, sodium azide, methyl methanesulfonate and 2-aminoanthracene, in Salmonella typhimurium TA98, Salmonella typhimurium TA100 and Escherichia coli WP2 uvrA strains. The chemical composition of the extract was analyzed by TLC and HPLC. Results Sisymbrium officinale showed to reduce the chemically-induced contractions of isolated guinea-pig trachea with major potency against leukotriene C 4 and histamine. The extract did not show any antibacterial activity. The Ames test showed a strong antimutagenic activity against 2-aminoanthracene, in Escherichia coli WP2 uvrA and in Salmonella typhimurium TA98 strains. The phytochemical study highlighted the presence of putranjivine, the glucosinolate marker of Sisymbrium officinale, and of proline. Conclusions The myorelaxant activity of Sisymbrium officinale offers a scientific basis to its use in traditional medicine. The strong antimutagenic effect suggests further studies to evaluate its possible chemopreventive activity.