RESUMEN
Some differently substituted 3-aryl-4,5-dihydropyrazoles-1-carbothioamides have been synthesised with the aim to investigate their monoamine oxidase inhibitory activity. The chemical structures of the compounds have been characterized by means of their IR, (1)H NMR, (13)C NMR spectroscopic data and elemental analyses. All the active compounds showed a selective activity towards the B isoform of the enzyme, regardless of the substitution on the heterocyclic ring. The inhibition of the enzymatic activity was measured on human recombinant MAO isoforms, expressed in baculovirus infected BTI insect cells. Docking experiments were carried out with the aim to rationalize the mechanism of inhibition of the most active and selective compound.
Asunto(s)
Inhibidores de la Monoaminooxidasa/química , Inhibidores de la Monoaminooxidasa/farmacología , Monoaminooxidasa/metabolismo , Pirazoles/química , Pirazoles/farmacología , Tioamidas/química , Tioamidas/farmacología , Animales , Línea Celular , Humanos , Insectos , Modelos Moleculares , Inhibidores de la Monoaminooxidasa/síntesis química , Unión Proteica , Pirazoles/síntesis química , Proteínas Recombinantes/antagonistas & inhibidores , Proteínas Recombinantes/metabolismo , Relación Estructura-Actividad , Tioamidas/síntesis químicaRESUMEN
The mechanisms involved in antiparasitic activity of the natural nonflavonoid polyphenol resveratrol (RESV) on the turbot (Psetta maxima) scuticoliate parasite Philasterides dicentarchi were investigated. At concentrations higher than 50microM, RESV caused significant inhibition of the in vitro growth of the ciliates, which was apparent on the third day of culture and, at the same concentration, RESV caused significant inhibition of O(2) consumption. RESV, at a concentration of 100microM, produced a significant increase in the production of intracellular reactive oxygen species (ROS), which were inhibited by the addition of 1mM of L (+) ascorbic acid. RESV (100microM) also caused significant inhibition of peroxidase, catalase and superoxide dismutase activities, but stimulated the activity of the redox regulating enzyme glutathione S-transferase. Confocal microscopy with the mitochondria-sensitive dye MitoTracker Orange CMTMRos revealed that RESV at concentrations higher than 50microM significantly increased the levels of fluorescence inside mitochondria and, at the same concentration, also caused an increase in the vacuolization of the trophozoites. The results obtained in the present study suggest that the inhibitory activity of RESV on the ciliate P. dicentrarchi is related to the induction of oxidative stress and to the inability of the parasite to eliminate ROS as a result of modified activity of antioxidant enzymes.
Asunto(s)
Antiprotozoarios/farmacología , Cilióforos/efectos de los fármacos , Peces Planos/parasitología , Estilbenos/farmacología , Animales , Antioxidantes/metabolismo , Apoptosis , Relación Dosis-Respuesta a Droga , Mitocondrias/efectos de los fármacos , Mitocondrias/metabolismo , Consumo de Oxígeno/efectos de los fármacos , Especies Reactivas de Oxígeno , Resveratrol , Factores de TiempoRESUMEN
Resveratrol (RESV; trans-3,5,4'-trihydroxystilbene), a phytoalexin that is produced by some plants, among other effects has well-known antioxidant, anti-inflammatory and immunomodulatory activities in mammals. In the present study, the effects of RESV on several functions of turbot, Psetta maxima (L.), kidney leucocytes (KLs) related to the innate and inflammatory responses were investigated. RESV exerted a dose-dependent inhibitory effect on the migratory response and on the production of reactive oxygen species in KL, after stimulation of the respiratory burst activity with phorbol myristate acetate (PMA). RESV also significantly inhibited the generation of the pro-inflammatory mediator prostaglandin E(2) (PGE(2)) in the supernatant of KL cultures stimulated with acidic sulphated polysaccharides (ASPs) from the seaweed Ulva rigida. The effects of the polyphenol on enzymatic activity and on myeloperoxidase (MPO) gene expression in neutrophils were also tested. It was found that RESV strongly inhibited intracellular and extracellular MPO activity, behaving as a noncompetitive and reversible inhibitor, and also induced a decrease in MPO mRNA levels in turbot neutrophils. These findings indicate that RESV exerts important modulatory effects on inflammatory responses in fish, and considering the importance of innate immunity in these vertebrates and the similarities with mammals, it may be possible to use fish for analysis of the effects of different substances on inflammatory responses.
Asunto(s)
Inmunidad Innata/efectos de los fármacos , Inflamación/metabolismo , Leucocitos/efectos de los fármacos , Estilbenos/farmacología , Animales , Movimiento Celular/efectos de los fármacos , Células Cultivadas , Dinoprostona/metabolismo , Peces Planos , Regulación de la Expresión Génica , Riñón/citología , Leucocitos/inmunología , Leucocitos/metabolismo , Peroxidasa/genética , Peroxidasa/metabolismo , Estallido Respiratorio/efectos de los fármacos , Resveratrol , Acetato de Tetradecanoilforbol/farmacologíaRESUMEN
BACKGROUND AND PURPOSE: We previously reported that agonist-induced rises in cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) in human umbilical vein endothelial cells (HUVEC) were inhibited after a short-term (2 min) pre-treatment with cAMP-elevating agents. The aim of this work was to study the effects of longer term (8 h) pre-treatment with dibutyryl-cAMP (db-cAMP) or rolipram, a specific inhibitor of phosphodiesterase-4 (PDE4), on [Ca(2+)](i), cAMP levels and PDE activity and expression in HUVEC. EXPERIMENTAL APPROACH: [Ca(2+)](i) changes were measured in isolated HUVEC by Fura-2 imaging. Intracellular cAMP levels and PDE4 activity were assessed by enzyme-immunoassay and radio-enzymatic assay, respectively. PDE expression was measured by northern and western blot analysis. KEY RESULTS: Long-term pre-treatment of HUVEC with rolipram or db-cAMP significantly increased ATP-, histamine- and thrombin-induced [Ca(2+)](i) rises. Short-term pre-treatment with rolipram was associated with an increase in cAMP, whereas long-term pre-treatment was associated with a decrease in cAMP. Long-term pre-treatment with rolipram or db-cAMP induced a significant increase in PDE4 activity and the expression of 74 kDa-PDE4A and 73 kDa-PDE4B was specifically enhanced. All these effects were suppressed by cycloheximide. CONCLUSIONS AND IMPLICATIONS: Our data suggest that sustained inhibition of PDE4 by rolipram induced an increase in PDE4 activity, possibly as a compensatory mechanism to accelerate cAMP degradation and that PDE4A and PDE4B were implicated in the regulation of [Ca(2+)](i). Thus, isozyme-specific PDE4 inhibitors might be useful as therapeutic agents in diseases where [Ca(2+)](i) handling is altered, such as atherosclerosis, hypertension and tolerance to beta-adrenoceptor agonists.
Asunto(s)
Señalización del Calcio/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Inhibidores de Fosfodiesterasa 4 , Inhibidores de Fosfodiesterasa/farmacología , 3',5'-AMP Cíclico Fosfodiesterasas/metabolismo , Northern Blotting , Western Blotting , Bucladesina/farmacología , Calcio/metabolismo , Calcio/fisiología , Línea Celular , AMP Cíclico/metabolismo , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 4/biosíntesis , Fosfodiesterasas de Nucleótidos Cíclicos Tipo 4/genética , Cicloheximida/farmacología , Interpretación Estadística de Datos , Colorantes Fluorescentes , Fura-2 , Histamina/farmacología , Humanos , Inhibidores de la Síntesis de la Proteína/farmacología , Rolipram/farmacologíaRESUMEN
Resveratrol (3,4',5-trihydroxystilbene, RESV) is a natural phenolic compound that exists as cis and trans isomers [c-RESV or (Z)-RESV and t-RESV or (E)-RESV, respectively]. t-RESV is a natural component of Vitis vinifera L. (Vitaceae), abundant in the skin of grapes (but not in the flesh) and in the leaf epidermis, and present in wines, especially red wines. In in vitro, ex vivo and in vivo experiments t-RESV exhibits a number of biological activities, including anti-inflammatory and anticarcinogenic properties. RESV also exists in wines as a cis isomer, which (unlike t-RESV) is not currently available commercially; as a result, little is known about this isomer's pharmacological activity. In this review, I will focus on the few comparative studies of the antioxidant effects of the two RESV isomers in different experimental models.
Asunto(s)
Antioxidantes/química , Antioxidantes/farmacología , Estilbenos/química , Estilbenos/farmacología , Animales , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Interferón gamma/farmacología , Kluyveromyces/patogenicidad , Peroxidación de Lípido/efectos de los fármacos , Lipopolisacáridos/farmacología , Macrófagos Peritoneales/efectos de los fármacos , Macrófagos Peritoneales/metabolismo , Ratones , Ratas , Especies Reactivas de Oxígeno/metabolismo , Resveratrol , EstereoisomerismoRESUMEN
This study investigated for the first time the effects of the cis isomer of RESV (c-RESV), a polyphenol present in red wine, on an array of genes whose expression is controlled by nuclear factor kappa B (NF-kappaB) and whose transcriptional activation is critical in a number of pathologies (including some cardiovascular diseases). In inflammatory peritoneal macrophages stimulated with lipopolysaccharide (LPS) and gamma interferon (IFN-gamma), c-RESV significantly blocked the expression of genes related to the REL/NF-kappaB/IkappaB family, adhesion molecules and acute-phase proteins; however, the greatest modulatory effect was obtained on the expression of genes related to the pro-inflammatory cytokines. c-RESV down-regulated the nuclear factor of kappa light chain gene enhancer in B-cells 1 (NFkappaBL1) gene product p105 and up-regulated the nuclear factor of kappa light chain gene enhancer in B-cells inhibitor alpha (IkappaBalpha) gene. c-RESV also significantly inhibited intercellular adhesion molecule-1 (ICAM-1) gene expression and the transmembrane receptors RIP (receptor TNFRSF) and TLR3 (toll-like receptor 7). At 100 muM, c-RESV significantly inhibited transcription of Scya2 (chemokine MCP-1), the chemokine RANTES (regulated on activation, normal T cell expressed and secreted), pro-inflammatory cytokines that attract monocyte-granulocyte cells such as M-CSF (colony-stimulating factor 1), GM-CSF (colony-stimulating factor 2) and G-CSF (colony-stimulating factor 3), the cytokine tumor growth factor beta (TGF-beta) and the extracellular ligand IL-1alpha. In contrast, c-RESV stimulated transcription of the pro-inflammatory cytokines IL-6 and tumor necrosis factor alpha (TNF-alpha), the extracellular ligand IL-1beta, and the IFN regulatory factor (IRF)-1. In conclusion, c-RESV has a significant modulatory effect on the NF-kappaB signaling pathway and, consequently, an important antioxidant role that may partially explain the cardioprotective effects attributed to long-term moderate red wine consumption.
Asunto(s)
Expresión Génica/efectos de los fármacos , Macrófagos Peritoneales/efectos de los fármacos , FN-kappa B/genética , Transducción de Señal/efectos de los fármacos , Estilbenos/farmacología , Animales , Células Cultivadas , Citocinas/biosíntesis , ADN/análisis , Ensayo de Inmunoadsorción Enzimática , Femenino , Macrófagos Peritoneales/metabolismo , Ratones , Ratones Endogámicos BALB C , Análisis de Secuencia por Matrices de Oligonucleótidos , Resveratrol , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transducción de Señal/genética , EstereoisomerismoRESUMEN
This study investigated the effects of an aqueous extract of the stem bark of Mangifera indica L. (Anacardiaceae; Vimang), which contains a defined mixture of components including polyphenols (principally mangiferin, MA), triterpenes, phytosteroids, fatty acids and microelements, on expression of inflammation mediators in inflammatory murine macrophages after stimulation in vitro with lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma). In vitro treatment with Vimang at 4 microg/ml reduced levels of NOS-2 mRNA and NOS-2, while treatment at 40 microg/ml also reduced levels of COX-2 mRNA, COX-2, and prostaglandin E2 (PGE2). Results suggested that MA is involved in these effects. In vitro treatment with Vimang at 40 microg/ml also inhibited mRNA levels of the proinflammatory cytokines interleukin 1beta (IL-1beta), tumor necrosis factor alpha (TNF-alpha) and colony-stimulating factor (GM-CSF), but did not affect mRNA levels of IL-6 or tumor growth factor-beta (TGF-beta). Extracellular release of TNF-alpha by inflammatory macrophages was inhibited by in vitro treatment with Vimang at the same concentrations that showed inhibition of TNF-alpha mRNA levels. The inhibition of TNF-alpha production appears to be at least partially attributable to MA. Vimang at 4 microg/ml decreased mRNA levels of nuclear factor-kappaB (NF-kappaB) but did not affect expression of the NF-kappaB inhibitor (IkappaB). These data indicate that the potent anti-inflammatory effects of Vimang are due to selective modulation of the expression of inflammation-related genes, leading to attenuation of macrophage activation.
Asunto(s)
Mediadores de Inflamación/antagonistas & inhibidores , Macrófagos Peritoneales/efectos de los fármacos , Mangifera , Extractos Vegetales/farmacología , Animales , Antiinflamatorios/farmacología , Supervivencia Celular/efectos de los fármacos , Citocinas/antagonistas & inhibidores , Citocinas/biosíntesis , Dexametasona/farmacología , Expresión Génica , Técnicas In Vitro , Mediadores de Inflamación/metabolismo , Interferón gamma/farmacología , Lipopolisacáridos/farmacología , Macrófagos Peritoneales/metabolismo , Ratones , Ratones Endogámicos BALB C , Corteza de la Planta/química , ARN Mensajero/antagonistas & inhibidores , ARN Mensajero/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Xantonas/farmacologíaRESUMEN
This study investigated the effects of trans-resveratrol (trans-3,4',5-trihydroxystilbene, RESV), a natural polyphenol from grapes with known antioxidant activity, on the respiratory-burst responses and phagocytic activity of rat macrophages. RESV at concentrations of 1-10 microM significantly and dose-dependently inhibited (a) the extracellular production of reactive oxygen intermediates (ROls) by resident peritoneal macrophages stimulated with phorbol 12-myristate 13-acetate (PMA) (a potent activator of protein kinase C, PKC) and (b) intracellular production of ROIs after opsonin-independent phagocytosis of Kluyveromyces lactis cells. Over the 10-100 microM concentration ranges, RESV likewise inhibited the production of reactive nitrogen intermediates (RNIs) by macrophages stimulated with thioglycollate. RESV concentrations above 10 microM also dose-dependently inhibited the phagocytosis of K. lactis cells. The results obtained demonstrate that RESV is a potent inhibitor of the antipathogen responses of rat macrophages and, thus, suggest that this agent may have applications in the treatment of diseases involving macrophage hyperresponsiveness.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Macrófagos/efectos de los fármacos , Estilbenos/farmacología , Animales , Relación Dosis-Respuesta a Droga , Kluyveromyces/efectos de los fármacos , Kluyveromyces/inmunología , Macrófagos/inmunología , Macrófagos/microbiología , Masculino , Nitritos/inmunología , Nitritos/metabolismo , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/inmunología , Especies Reactivas de Oxígeno/metabolismo , ResveratrolRESUMEN
The toxic effects derived from overproduction of oxygen radicals [reactive oxygen species (ROS)] by immune cells can be partially abolished by the antioxidant activities of plant polyphenols. In the present study, we investigated the antioxidant action of a catechin, (-)-epigallocatechin-3-gallate (EGCG), on the respiratory-burst responses of rat peritoneal macrophages. EGCG at concentrations of 50-200 microM blocked the production of nitric oxide by macrophages stimulated in vivo with sodium thioglycollate then 5 days later in vitro with lipopolysaccharide and gamma-interferon. At 1-100 microM, EGCG also inhibited the extracellular liberation of oxygen radicals by resident peritoneal macrophages stimulated with the protein kinase C activator phorbol 12-myristate 13-acetate (PMA). At low concentrations (1-5 microM), EGCG increased the reduction of nitro blue tetrazolium (NBT) by the superoxide anions generated in the non-enzymatic system NADH/PMS, acting as a pro-oxidant agent, while at concentrations above 10 microM, EGCG acts as a scavenger of superoxide anions. These results show that EGCG is capable of modulating ROS production during the respiratory burst of rat peritoneal macrophages by acting as a superoxide anion scavenger. EGCG may therefore be useful in the prevention and treatment of diseases due to increased free radical production.
Asunto(s)
Antioxidantes/farmacología , Catequina/análogos & derivados , Catequina/farmacología , Macrófagos Peritoneales/efectos de los fármacos , Estallido Respiratorio/efectos de los fármacos , Animales , Células Cultivadas , Macrófagos Peritoneales/metabolismo , Masculino , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Estallido Respiratorio/fisiologíaRESUMEN
In this work, the potential vasorelaxant activity of (+)-nantenine, an alkaloid isolated from Platycapnos spicata, was studied for the first time in rat aorta. (+)-Nantenine (3 - 30 microM) totally relaxed, in a concentration-dependent manner and with almost equal effectiveness, the contractions induced by noradrenaline (NA) or by a high KCl concentration (60 mM) in intact rat aortic rings. Mechanical removal of endothelium and/or pretreatment of aorta rings with glibenclamide (10 microM) or tetraethylammonium (TEA, 2 mM) did not significantly modify the vasorelaxant effects of this aporphine alkaloid. In the experiments in Ca(2+)-free medium, (+)-nantenine (10 microM) had no effect on caffeine-induced contractions. Furthermore, in the studies with radiolabelled Ca(2+), (+)-nantenine (3 - 30 microM) did not modify the basal uptake of (45)Ca(2+) but decreased, in a concentration-dependent fashion, the influx of (45)Ca(2+) induced by NA and KCl in endothelium-containing and endothelium-denuded rat aortic rings. In addition, (+)-nantenine (3 - 30 microM) was ineffective to scavenge superoxide anion (O(2) (-)) radicals generated by the hypoxanthine (HX)-xanthine oxidase (XO) system and/or to inhibit XO activity. These results indicate that: a) the vasorelaxant effects of (+)-nantenine in rat aorta are due, at least in part, to a blockage of Ca(2+) influx through transmembrane calcium channels, b) the activation of ATP-sensitive K(+) channels (K(ATP)) and large conductance Ca(2+)-activated K(+) channels (K(Ca)) present in smooth muscle cells, the presence (integrity) of endothelial system, an inhibitory action on XO enzymatic activity and/or O(2)(-) radicals scavenging properties are not involved in the vascular effects of (+)-nantenine in rat aorta described above.
Asunto(s)
Aorta/efectos de los fármacos , Aporfinas/farmacología , Papaveraceae , Vasodilatadores/farmacología , Animales , Aorta/fisiología , Aporfinas/química , Cafeína/farmacología , Calcio/metabolismo , Bloqueadores de los Canales de Calcio/farmacología , Relación Dosis-Respuesta a Droga , Técnicas In Vitro , Masculino , Extractos Vegetales/química , Extractos Vegetales/farmacología , Ratas , Ratas Wistar , Superóxidos/metabolismo , Vasoconstricción/efectos de los fármacos , Vasodilatación/efectos de los fármacos , Xantina Oxidasa/metabolismoRESUMEN
The synthesis of a novel series of 2,3-dihydro-3-oxo-4H-thieno[3, 4-e][1,2,4]thiadiazine 1,1-dioxides and their pharmacological evaluation as drugs with effects on the rat cardiovascular system are described. The compounds under study were synthesized via Curtius rearrangement of appropriate sulfamoylacylazides which, in turn, were prepared from known starting materials. In isolated rat portal vein, these thienothiadiazines, like verapamil and diazoxide, inhibited the spontaneous motility produced by KCl (20 mM). In addition, the new compounds, like verapamil and unlike diazoxide, also exhibited inhibitory effects in the same preparation when the cell membrane was depolarized by an increased extracellular KCl concentration (80 mM) and, consequently, the membrane potential approached a level close to the K(+) equilibrium potential. Further characterization of this inhibitory activity led to the identification of a selective inhibitory effect of the new compounds on KCl (80 mM)-induced 45Ca(2+) uptake in the same vascular tissue. When tested in vivo (anaesthetized normotensive rats), acute administration of verapamil, diazoxide and some of the most in vitro potent compounds in 45Ca(2+) uptake experiments produced a gradual, dose-dependent and sustained decrease in diastolic arterial blood pressure, devoid of cardiac effects. These results suggest that, like verapamil, the cardiovascular effects produced by the new thienothiadiazines seem to be due, at least in part, to a blockade of transmembrane voltage-dependent calcium channels present in vascular smooth muscle cells and not to an activation of ATP-sensitive K(+) channels. Compounds 5b, 5e and 5i have been selected for further studies as antihypertensive agents.
Asunto(s)
Bloqueadores de los Canales de Calcio/síntesis química , Bloqueadores de los Canales de Calcio/farmacología , Tiazinas/síntesis química , Tiazinas/farmacología , Animales , Bloqueadores de los Canales de Calcio/química , Evaluación de Medicamentos , Técnicas In Vitro , Activación del Canal Iónico , Dosificación Letal Mediana , Masculino , Estructura Molecular , Ratas , Ratas Endogámicas WKY , Análisis Espectral , Tiazinas/química , Venas/efectos de los fármacosRESUMEN
The in vivo and in vitro cardiovascular effects of the novel 5-HT2A/alpha1/H1 antagonist ketanserin analogues QF 0303B, QF 0307B, QF 0311B, QF 0313B were studied in anaesthetized normotensive rats (ANR) and in isolated rubbed rat aorta (IRRA). In ANR, 0.2 mg x kg(-1) i.v. of each compound produced a rapid, remarkable but short-lasting fall in mean arterial blood pressure (MAP) accompanied by bradycardia. All compounds significantly modified the pressor effects induced by 5-hydroxytryptamine (5-HT) and noradrenaline (NA). In IRRA, the compounds inhibited NA- and 5-HT-induced contractions in a competitive fashion. Furthermore, the analogues displayed lower H1-antagonist activity than ketanserin. Compounds tested showed low 5-HT2B affinity and no activity at muscarinic, nicotinic, or 5-HT3 receptors, nor any marked ability to produce smooth muscle relaxation via calcium entry blockade. There is a significant correlation between hypotension reached and inhibition of the 5-HT-induced pressor responses (but not for NA). A certain degree of correlation was observed between hypotensive effect endurance vs. alpha1-adrenoceptor blockade (but not for serotonin). These results indicate that in this series the brief hypotensive activity in ANR is attributed to a 5-HT1A receptor blockade and the duration of the effect is better attributed to an alpha1 adrenoceptor blockade.
Asunto(s)
Antihipertensivos/farmacología , Ketanserina/farmacología , Receptores Adrenérgicos alfa 1/metabolismo , Receptores de Serotonina/metabolismo , Antagonistas Adrenérgicos alfa/farmacología , Animales , Antihipertensivos/química , Aorta , Cobayas , Íleon/efectos de los fármacos , Íleon/metabolismo , Ketanserina/análogos & derivados , Ketanserina/química , Masculino , Músculos/efectos de los fármacos , Músculos/metabolismo , Cloruro de Potasio/farmacología , Ratas , Ratas Sprague-Dawley , Receptor de Serotonina 5-HT2A , Receptor de Serotonina 5-HT2B , Receptores Adrenérgicos alfa 1/efectos de los fármacos , Receptores Histamínicos H1/efectos de los fármacos , Receptores Histamínicos H1/metabolismo , Receptores de Serotonina/efectos de los fármacos , Receptores de Serotonina 5-HT3 , Antagonistas de la Serotonina/farmacología , Vasoconstricción/efectos de los fármacosRESUMEN
In this work, we report for first time that: (1) low doses of ketanserin (0.2 mg/kg) produce a transient hypotensive response in anaesthetized rats, which is basically due to the blockade of 5-hydroxytryptamine (2A) (5-HT)2A receptors, whereas high doses (1 mg/kg) of ketanserin cause a sustained hypotension also mediated by the blockage of alpha1-adrenergic receptors; (2) the in vitro vasorelaxant action of high concentrations of ketanserin (>10 microM) involves Ca2+ antagonism, which may also be responsible, at least in part, for the inhibition of high-K+-induced 45Ca2+ uptake, the inhibition of Ca2+-induced contractions in initially Ca2+-free high-K+ medium, and the negative chronotropic effects on isolated atria. This Ca2+ antagonistic activity does not seem to contribute to the in vivo cardiovascular effects of ketanserin at therapeutic doses.
Asunto(s)
Hemodinámica/efectos de los fármacos , Ketanserina/farmacología , Antagonistas de la Serotonina/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , Calcio/metabolismo , Relación Dosis-Respuesta a Droga , Frecuencia Cardíaca/efectos de los fármacos , Técnicas In Vitro , Masculino , Norepinefrina/farmacología , Ratas , Ratas Sprague-Dawley , Serotonina/farmacologíaRESUMEN
PURPOSE: The syntheses and evaluation for cardiovascular activity in the rat of both enantiomers of a verapamil analog in which the cyano group has been replaced by hydroxyl. METHODS: (+)- and (-)-alpha-[3-[[2-(3,4-Dimethoxyphenyl)ethyl]methylamino]propyl]- 3,4-dimethoxy-alpha-(1-methyl ethyl)benzyl alcohol were prepared from chiral sulfoxides produced by microbial biotransformations using Mortierella isabellina ATCC 42613 or Helminthsporium species NRRL 4671, and were examined for hypotensive and calcium antagonist activity using anaesthetized normotensive rats and isolated rat aorta and atria. RESULTS: The analogs showed a pharmacological profile similar to that exhibited by verapamil, possessing a remarkable hypotensive activity, accompanied by a significant bradycardia, in anaesthetized normotensive rats. In vitro, these analogs displayed clear inhibitory effects: in isolated rat aorta they inhibited, in a concentration-dependent fashion, the contractions and 45Ca2+ uptake induced by norepinephrine and high KCl, and in isolated rat atria the analogs considerably decreased the rate of contraction (negative chronotropic effects). No significant differences between the quantitative cardiovascular effects produced by the two enantiomers of the verapamil analogs were observed. CONCLUSIONS: The results suggest that, like that of verapamil, the cardiovascular activity exhibited by the new compounds seems to be due, at least in part, to a blockage of transmembrane calcium channels present in vascular smooth muscle cells.
Asunto(s)
Bloqueadores de los Canales de Calcio/farmacología , Hipotensión/inducido químicamente , Vasoconstricción/efectos de los fármacos , Verapamilo/farmacología , Animales , Aorta/efectos de los fármacos , Aorta/fisiología , Función Atrial/efectos de los fármacos , Biotransformación , Bloqueadores de los Canales de Calcio/síntesis química , Relación Dosis-Respuesta a Droga , Helminthosporium/metabolismo , Técnicas In Vitro , Masculino , Mortierella/metabolismo , Ratas , Ratas Endogámicas WKY , Estereoisomerismo , Verapamilo/análogos & derivados , Verapamilo/síntesis químicaRESUMEN
1. For several years we have been working on the synthesis of modified hydrazinopyridazines which have proved to possess remarkable vasorelaxant and antihypertensive activity. We now report the vasodilator effects of three novel 6-aryl-5-piperidino-3-hydrazinopyridazines (1a, 1b and 1c), structurally related to the well-known antihypertensive drug hydralazine. 2. Hydralazine and the new hydrazinopyridazines relaxed, in a concentration-dependent and nonspecific way, the contractions elicited by noradrenaline or a high K+ concentration in rat aortic rings with or without endothelium. According to the IC50 (50% inhibitory concentrations) values obtained, the vasorelaxant potency of the new compounds was greater than that of hydralazine. 3. In a Ca2+-free medium, the contractions provoked by noradrenaline or caffeine were significantly inhibited by the new hydrazinopyridazines and by hydralazine. 4. Hydralazine and the novel molecules did not significantly modify basal, noradrenaline- or K+-induced 45Ca2+ uptake. 5. These results suggest that 1a, 1b and 1c have an endothelium-independent vasorelaxant activity greater than that of hydralazine in isolated rat aortic rings, which seems not to be mediated by a blockade of transmembrane Ca2+ movements through specific channels. This effect could be due, at least in part, to an intracellular mechanism of action.
Asunto(s)
Aorta Torácica/efectos de los fármacos , Hidralazina/farmacología , Piridazinas/farmacología , Vasodilatadores/farmacología , Animales , Aorta Torácica/metabolismo , Aorta Torácica/fisiología , Cafeína/farmacología , Calcio/metabolismo , Técnicas In Vitro , Masculino , Contracción Muscular , Norepinefrina/farmacología , Piridazinas/química , Ratas , Ratas Endogámicas WKY , Relación Estructura-Actividad , Vasodilatadores/químicaRESUMEN
In this work, the potential vasorelaxant activity of centaurein and centaureidin, two flavonoids from Centaurea corcubionensis, were studied for the first time in rat aorta. Centaureidin (10 microM-0.1 mM) totally relaxed, in a concentration-dependent manner and with almost equal effectiveness, the contractions induced by NA (IC50 = 16.7 +/- 1.9 microM) or by a high K+ concentration (IC50 = 16.1 +/- 3.1 microM) in intact rat aortic rings. Mechanical removal of endothelium did not significantly modify the vasoralexant effects of this flavone (IC50 = 20.8 +/- 2.4 microM for NA; IC50 = 21 +/- 2.9 microM for K+). On the other hand, centaurein (1 microM-0.1 mM) had no effect on NA- and high K(+)-induced contractions in rubbed and intact rat aortic rings. These results indicate that substitution by glucose in the chemical structure of centaureidin leads to the loss of its vasodilator activity.
Asunto(s)
Aorta/fisiología , Flavonoides/aislamiento & purificación , Glucósidos/aislamiento & purificación , Músculo Liso Vascular/fisiología , Plantas Medicinales , Vasodilatación/fisiología , Vasodilatadores/aislamiento & purificación , Animales , Aorta/efectos de los fármacos , Femenino , Flavonoides/química , Flavonoides/farmacología , Glucósidos/química , Glucósidos/farmacología , Técnicas In Vitro , Estructura Molecular , Contracción Muscular/efectos de los fármacos , Músculo Liso Vascular/efectos de los fármacos , Ratas , Ratas Wistar , Vasodilatación/efectos de los fármacos , Vasodilatadores/química , Vasodilatadores/farmacologíaRESUMEN
1. In this work, the cardiovascular effects of HPS-10, a new vasodilator agent, were studied in rats. 2. In conscious normotensive rats, oral administration of HPS-10 (4-9 mg kg-1) produced a dose-related and long-lasting fall in systolic arterial blood pressure (ED30 of 5.32 mg kg-1), accompanied by an increase in heart rate (ED30 of 8.43 mg kg-1). This tachycardia was totally inhibited by pretreatment with (+/-)-propranolol (10 mg kg-1, p.o.). 3. In anaesthetized normotensive rats, HPS-10 (0.3-0.6 mg kg-1, i.v.) produced a gradual, dose-dependent and sustained decrease in systolic, diastolic and mean arterial pressure (MAP) (ED30 for MAP of 0.41 mg kg-1, i.v.), accompanied by a significant bradycardia at high doses (> 0.4 mg kg-1; ED20 of 0.61 mg kg-1, i.v.). HPS-10 (0.5 mg kg-1, i.v.) did not modify the positive chronotropic effects induced by intravenous administration of noradrenaline (NA; 5 micrograms kg-1), angiotensin II (AII; 0.2 microgram kg-1) and nicotine (200 micrograms kg-1) but markedly inhibited the hypertensive response produced by these agents. 4. In rat isolated rubbed aorta, HPS-10 (0.1-1 mM) non-competitively and with almost equal effectiveness antagonized the contractions induced by NA, AII (in normal Krebs solution) and Ca2+ (in depolarizing Ca(2+)-free high-K+ 50 mM solution). In the experiments in Ca(2+)-free medium, HPS-10 (1 mM) considerably inhibited the contractions induced by NA, AII and caffeine in rat aorta. 5. Furthermore, in the studies with radioactive Ca2+, HPS-10 (1 mM) did not modify the basal uptake of 45Ca2+ but strongly decreased the influx of 45Ca2+ induced by NA, AII and K+ in rat aortic rings. 6. In rat isolated atria, HPS-10 (1 mM) produced a positive inotropic/negative chronotropic effect. 7. HPS-10 (0.3 mM) significantly inhibited the sustained and transient Ba2+ inward current (IBa) recorded in whole-cell clamped rat aortic myocytes. 8. These results indicate that the non-selective vasorelaxant effects of HPS-10 in rat aortic rings can be attributed to transmembrane Ca(2+)-antagonist activity and an intracellular action on smooth muscle cells. The direct vasodilator action of HPS-10 observed in rat isolated aorta may be responsible for the HPS-10 hypotensive activity in anaesthetized normotensive rats.
Asunto(s)
Presión Sanguínea/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Hidrazinas/farmacología , Piridazinas/farmacología , Vasodilatadores/farmacología , Angiotensina II/farmacología , Animales , Calcio/metabolismo , Técnicas In Vitro , Masculino , Ratones , Contracción Miocárdica/efectos de los fármacos , Norepinefrina/farmacología , Ratas , Ratas Wistar , Vasoconstricción/efectos de los fármacosRESUMEN
The effects of NG-nitro-L-arginine (L-NNA) on mean arterial pressure and the effects of both L-NNA and methylene blue on isolated aorta tone, were studied in order to elucidate potential alterations in vasodilator resting nitric oxide (NO) tone in genetic hypertension. L-NNA produced a significantly greater increase of mean arterial pressure in spontaneously hypertensive rats (SHR) than in Wistar Kyoto (WKY) rats; in both cases, L-arginine completely inhibited the L-NNA hypertensive effect. Neither ganglion blockade with hexamethonium nor cyclooxygenase inhibition with indomethacin significantly modified the effect of L-NNA in both rat strains. In intact aorta rings, after submaximally contraction with KCI (25 mM), both L-NNA and methylene blue induced strong dose-dependent contractions. The maximum contractions were, however, significantly greater in WKY rats than in SHR. The mechanical elimination of endothelium markedly inhibited both L-NNA and methylene blue maximum contractions. In intact rings, L-arginine completely inhibited the L-NNA effects in both rat strains; in rubbed rings, the L-arginine inhibitory effects were strong in WKY rats but not important and erratic in SHR. L-Arginine had no effect on the contractions induced only by KCI in any of the preparations. In WKY rat-rubbed rings, sodium nitroprusside was significantly more effective in relaxing the contractions in response to 25 mM KCI than the contractions in response to methylene blue. These results indicate that contractions induced by L-NNA and methylene blue in isolated aorta are principally due to the inhibition of an important endothelial resting vasodilator NO tone. They also show that hypertension reduces the resting vasodilator NO tone in isolated rat aorta, in spite of enhancing the total vasodilator NO tone in anaesthetized rat.
Asunto(s)
Óxido Nítrico/fisiología , Resistencia Vascular/fisiología , Animales , Aorta Torácica/efectos de los fármacos , Aorta Torácica/fisiología , Presión Sanguínea/efectos de los fármacos , Hexametonio/farmacología , Técnicas In Vitro , Indometacina/farmacología , Masculino , Azul de Metileno/farmacología , Fenilefrina/farmacología , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Vasoconstrictores/farmacología , Vasodilatadores/farmacología , omega-N-Metilarginina/farmacologíaRESUMEN
Ca2+ plays an important role in the contraction of skeletal, cardiac, and smooth muscle, as well as in a number of important processes, such as secretion and neuronal activity. In this review, I focus on the various mechanisms by which cytosolic Ca2+ concentration is regulated in vascular smooth muscle, in the resting state and during activation. Particular attention is paid to the calcium pumps of the plasmalemma and the sarcoplasmic reticulum, to the inositol 1,4,5-trisphosphate- and ryanodine-sensitive calcium channels of the sarcoplasmic reticulum, and to voltage-dependent and voltage-independent calcium channels of the plasmalemma.
Asunto(s)
Canales de Calcio/metabolismo , Calcio/metabolismo , Músculo Liso Vascular/metabolismo , Retículo Sarcoplasmático/metabolismo , Canales de Calcio/fisiología , Humanos , Contracción Muscular/efectos de los fármacos , Músculo Liso Vascular/efectos de los fármacosRESUMEN
1. The cardiovascular and vasorelaxant effects of (+)-glaucine and of a semisynthetic derivative (N-carbethoxysecoglaucine) were studied in rats. 2. N-carbethoxysecoglaucine did not modify either systolic arterial pressure or heart rate values in conscious (25 mg kg-1, p.o.) and anaesthetized normotensive rats (5 mg kg-1, i.v.). Furthermore, this compound showed no activity in the experiments carried out on rat isolated aorta [contractility and 45Ca2+ influx assays (5 microM)] and did not modify the rate and force of contraction in rat isolated atria (5 microM). 3. In conscious normotensive rats, oral administration of (+)-glaucine (25 mg kg-1) did not modify either systolic arterial pressure or heart rate. 4. In anaesthetized normotensive rats, (+)-glaucine (5 mg kg-1, i.v.) produced a remarkable fall in mean arterial pressure (MAP) accompanied by a significant decrease in heart rate. In the same preparation, (+)-glaucine (5 mg kg-1, i.v.) did not modify the cardiovascular effects induced by noradrenaline (NA) (5 micrograms kg-1) and 5-hydroxytryptamine (5-HT) (300 micrograms kg-1) but markedly inhibited those induced by nicotine (200 micrograms kg-1). 5. In isolated intact aorta of rat, (+)-glaucine (0.15-5 microM) competitively inhibited the contractions induced by NA (with a pA2 value of 7.14) and non-competitively those induced by 5-HT (in normal Krebs solution) and Ca2+ (in depolarizing Ca(2+)-free high-K+ 50 mM solution), with depression of the maximal response and with pD2 values of 5.56 and 5.26, respectively. 6. In experiments in Ca2+-free medium, (+)-glaucine (3 microM) inhibited the contractions induced by NA and had no effect on either 5-HT- or caffeine-induced contractions.7. Furthermore, in the experiments with radioactive Ca2+, (+)-glaucine (3 microM) did not modify the basal uptake of 45Ca2+ but strongly inhibited the influx of 45Ca2+ induced by NA, 5-HT and K+.8. (+)-Glaucine (5microM) had no effect on rate and force of contraction in rat isolated atria.9. These results indicate that: (a) the cardiovascular effects (hypotension and bradycardia) of (+)-glaucine in anaesthetized normotensive rats (5 mg kg-1) may be due, at least in part, to a ganglioplexic effect; (b) the vasorelaxant action of ( + )-glaucine (0.15-5 microM) in rat isolated aorta can be attributed to an alpha1-adrenoceptor blocking property (which may explain its inhibition of noradrenaline-induced 45Ca2+influx and contractions in normal Krebs solution and noradrenaline-induced contractions in Ca2+-free medium) and to a Ca2+-antagonist activity (which may be responsible, at least in part, for the inhibition of 45Ca2+ uptake induced by NA, 5-HT and K+ and the contractions induced by both NA and 5-HT in normal Krebs solution and by Ca2+ in Ca2+-free high-K+ medium) and (c) there is no correlation between the mechanisms of action observed for (+ )-glaucine in vivo and in vitro, which suggests that the vasorelaxant activity of this alkaloid does not contribute to its hypotensive activity.