RESUMO
Torsade de pointes is a rare but potentially fatal ventricular arrhythmia associated with drug-induced delayed repolarization and prolongation of the QT interval. To determine if the arrhythmogenic potential of noncardiac drugs can be assessed in vitro, we evaluated the effects of 12 drugs on the action potential duration (APD) of cardiac Purkinje fibers and compared results with clinical observations. APD changes in canine and porcine fibers were evaluated under physiologic conditions (37 degrees C, [K+]0 = 4 mM) using standard microelectrode techniques. Six of seven drugs associated with QT prolongation or torsade de pointes in man (cisapride, erythromycin, grepafloxacin, moxifloxacin, sertindole, and sotalol) affected concentration-dependent prolongation of the APD in canine fibers during slow stimulation (2-s basic cycle length), attaining greater than 15% prolongation at high concentrations (> or = 10-fold clinically encountered plasma levels). Each of five drugs not linked clinically to QT prolongation and torsade de pointes (azithromycin, enalaprilat, fluoxetine, indomethacin, and pinacidil) failed to attain 15% prolongation, with fluoxetine, indomethacin, and pinacidil abbreviating the APD. Drugs eliciting the greatest prolongation also demonstrated prominent reverse rate-dependent effects. The antihistamine terfenadine (linked to dose-dependent QT prolongation and torsade de pointes clinically) only minimally prolonged the APD in canine and porcine fibers (and exerted no effect on midmyocardial fibers from left ventricular free wall) at supratherapeutic concentrations. On the basis of concentration-dependent APD prolongation and reverse rate-dependent effects, this Purkinje fiber model detects six of seven drugs linked clinically to acquired long QT syndrome and torsade de pointes, and clears each of five drugs not associated with repolarization abnormalities (overall 92% accuracy), validating the utility of this Purkinje fiber model in the preclinical evaluation of QT prolongation and proarrhythmic risk by noncardiac drugs.
Assuntos
Potenciais de Ação/efeitos dos fármacos , Arritmias Cardíacas/induzido quimicamente , Compostos Aza , Fluoroquinolonas , Síndrome do QT Longo/fisiopatologia , Ramos Subendocárdicos/efeitos dos fármacos , Quinolinas , Potenciais de Ação/fisiologia , Animais , Anti-Infecciosos/farmacologia , Arritmias Cardíacas/fisiopatologia , Cisaprida/farmacologia , Cães , Eritromicina/farmacologia , Feminino , Imidazóis/farmacologia , Técnicas In Vitro , Indóis/farmacologia , Síndrome do QT Longo/induzido quimicamente , Masculino , Modelos Biológicos , Moxifloxacina , Perfusão , Piperazinas/farmacologia , Ramos Subendocárdicos/fisiologia , Sotalol/farmacologia , Suínos , Torsades de Pointes/induzido quimicamente , Torsades de Pointes/fisiopatologiaRESUMO
Sub-acute and chronic bronchitis (SACB) are among the least well studied major medical problems that exist today. While much research and novel drug discovery have focused on asthma, comparatively little has been performed on chronic bronchitis, the fourth or fifth most frequent disease (4 to 6% of the population over 45 years of age), or on sub-acute bronchitis, the persistent symptoms of a respiratory infection which is the major reason Americans visit a physician (20%). This lack of attention is largely due to difficulties associated with modeling the pathophysiology of SACB in vitro, in situ, in organs or in animals as well as the presumption that drugs developed for asthma would also be effective in SACB. Data with bronchodilators (anti-cholinergics versus beta2-adrenergic agonist) and corticosteroids have strongly dismissed that premise. In this review of potential novel mechanistic targets directed specifically at SACB, an emphasis is given to recent data, gathered most convincingly from tissues and patients with cystic fibrosis, that have led to an appreciation of the critical role respiratory epithelial dysfunction plays in the pathophysiology and symptoms of these diseases. Mechanistic targets that restore (normalize or accelerate) airway 'cleansing' (enhance host defense) by accelerating mucociliary clearance are described and given preference over anti-inflammatory mechanisms that could further impair host defense.
Assuntos
Corticosteroides/uso terapêutico , Bronquite/tratamento farmacológico , Bronquite/fisiopatologia , Broncodilatadores/uso terapêutico , Mucosa Respiratória/fisiologia , Antioxidantes/metabolismo , Antioxidantes/uso terapêutico , Bronquite/microbiologia , Fibrose Cística/fisiopatologia , Humanos , Inflamação/tratamento farmacológico , Inflamação/fisiopatologia , Modelos Biológicos , Muco/metabolismo , Agonistas do Receptor Purinérgico P2 , Receptores Purinérgicos P2Y2 , Mucosa Respiratória/efeitos dos fármacos , Bloqueadores dos Canais de SódioAssuntos
Asma/tratamento farmacológico , Broncoconstritores/farmacologia , Modelos Animais de Doenças , Doenças dos Macacos/tratamento farmacológico , Pneumonia Viral/tratamento farmacológico , Infecções por Vírus Respiratório Sincicial/tratamento farmacológico , Doença Aguda , Alérgenos/farmacologia , Animais , Asma/induzido quimicamente , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Doença Crônica , Feminino , Macaca fascicularis , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pneumonia , Eosinofilia Pulmonar/tratamento farmacológico , Vírus Sinciciais RespiratóriosRESUMO
This study was designed to quantify the long-term contribution of endogenous endothelin-1 (ET-1) and ET(A) receptors to the regulation of arterial pressure under normal conditions in nonhuman primates. Therefore, mean arterial pressure (MAP) and heart rate were measured 24 h/day with the use of telemetry techniques in conscious cynomolgus monkeys under control conditions, during administration of an ET(A) selective receptor antagonist (ABT-627; 5 mg/kg, 2 times a day by mouth, 4 days), and a 6-day posttreatment period. Systemic ET(A) blockade reduced MAP (24 h) from 89 +/- 3 to 82 +/- 2 and 79 +/- 2 mmHg on days 1 and 4, respectively. Subsequently, MAP remained suppressed for 3 days posttreatment. Heart rate increased from 111 +/- 5 to 122 +/- 4 and 128 +/- 6 beats/min on days 1 and 4 of ABT-627, respectively, and remained above control for 3 days posttreatment. Plasma ET-1 concentration increased from 1.0 +/- 0.3 to 1.9 +/- 0.4 pg/ml in response to ABT-627 (day 4) but decreased to control values 4 days posttreatment. These data demonstrate a physiologically important role for endogenous ET-1 and ET(A) receptors in the long-term regulation of arterial pressure and plasma ET-1 levels in the conscious nonhuman primate.
Assuntos
Pressão Sanguínea/fisiologia , Endotelina-1/fisiologia , Homeostase , Receptores de Endotelina/fisiologia , Animais , Atrasentana , Antagonistas dos Receptores de Endotelina , Endotelina-1/sangue , Frequência Cardíaca , Hematócrito , Cinética , Macaca fascicularis , Masculino , Potássio/sangue , Pirrolidinas/farmacologia , Receptor de Endotelina A , Sódio/sangueRESUMO
A series of bis(trifluoromethyl)pyrazoles (BTPs) has been found to be a novel inhibitor of cytokine production. Identified initially as inhibitors of IL-2 synthesis, the BTPs have been optimized in this regard and even inhibit IL-2 production with a 10-fold enhancement over cyclosporine in an ex vivo assay. Additionally, the BTPs show inhibition of IL-4, IL-5, IL-8, and eotaxin production. Unlike the IL-2 inhibitors, cyclosporine and FK506, the BTPs do not directly inhibit the dephosphorylation of NFAT by calcineurin.
Assuntos
Quimiocinas CC , Proteínas de Ligação a DNA/metabolismo , Proteínas Nucleares , Inibidores da Síntese de Proteínas/síntese química , Pirazóis/síntese química , Fatores de Transcrição/metabolismo , Animais , Asma/tratamento farmacológico , Divisão Celular , Quimiocina CCL11 , Técnicas de Química Combinatória , Ciclosporina/farmacologia , Citocinas/antagonistas & inibidores , Citocinas/biossíntese , Genes Reporter , Haplorrinos , Humanos , Imunossupressores/síntese química , Imunossupressores/química , Imunossupressores/farmacologia , Técnicas In Vitro , Interleucina-2/antagonistas & inibidores , Interleucina-2/biossíntese , Interleucina-4/antagonistas & inibidores , Interleucina-4/biossíntese , Interleucina-5/antagonistas & inibidores , Interleucina-5/biossíntese , Interleucina-8/antagonistas & inibidores , Interleucina-8/biossíntese , Células Jurkat , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Luciferases/genética , Fatores de Transcrição NFATC , Inibidores da Síntese de Proteínas/química , Inibidores da Síntese de Proteínas/farmacologia , Pirazóis/química , Pirazóis/farmacologia , RatosRESUMO
Endothelin-1 (ET-1), a potent vasoactive and mitogenic peptide, has been implicated in a number of cardiovascular diseases, including congestive heart failure, neointimal hyperplasia associated with restenosis, and hypertension. The vasoconstriction induced by ET-1 is thought to be mediated mainly by its action on ET(A) receptors on vascular smooth muscle cells. Recent studies have indicated that vasoconstriction also may be mediated by stimulation of an ET(B)-receptor subtype. Increased use of the pig as a cardiovascular model prompted us to examine the receptor profile in this species using ABT-627, a potent, nonpeptide antagonist of the ET(A) receptor. The precursor to ET-1, big ET-1 (0.02 nmol/kg/min), was infused intravenously in domestic swine, resulting in a sustained increase in mean blood pressure of 38 +/- 3 mm Hg. After stabilization of the pressor response, ABT-627 (0.1-10 microg/kg/min) or vehicle was infused for 30 min. Whereas vehicle infusion had no appreciable effect, a dose-related reversal of the pressor response to big ET-1 (11-100%) was observed by the end of the ABT-627 infusion. Blood samples were assayed for plasma concentrations of ABT-627; peak levels ranged from 9 +/- 2 to 937 +/- 168 ng/ml. In a separate group of pigs, the highest dose of ABT-627 produced only a modest reversal of the hypertensive response to an infusion of angiotensin II (300 ng/kg/min). Additional results indicate that the vasoconstrictor effects produced by sarafotoxin 6C (0.03 and 0.3 nmol/kg), an agonist of the ET(B) receptor, are not blocked by treatment with ABT-627 (10 microg/kg/min). However, complete blockade of the S6C response could be achieved using the ET(B) antagonist, A-192621 (0.33 mg/kg/min). Our results define the dose-response relation for the ET(A)-receptor antagonist ABT-627 in the vasculature of the domestic pig and suggest the presence of an ET(B)-receptor subtype that mediates vasoconstriction in this species.
Assuntos
Receptores de Endotelina/fisiologia , Vasoconstrição/fisiologia , Angiotensina II/farmacologia , Animais , Animais Domésticos , Atrasentana , Pressão Sanguínea/efeitos dos fármacos , Relação Dose-Resposta a Droga , Antagonistas dos Receptores de Endotelina , Endotelina-1 , Endotelinas/sangue , Endotelinas/farmacologia , Feminino , Artéria Femoral/efeitos dos fármacos , Artéria Femoral/fisiologia , Precursores de Proteínas/sangue , Precursores de Proteínas/farmacologia , Pirrolidinas/sangue , Pirrolidinas/farmacologia , Receptor de Endotelina A , Receptor de Endotelina B , Suínos , Fatores de Tempo , Vasoconstritores/farmacologia , Venenos de Víboras/farmacologiaRESUMO
ABT-229 (8,9-anhydro-4"-deoxy-3'-N-desmethyl-3'-N-ethylerythromycin B-6,9-hemiacetal), a synthetic derivative of erythromycin (ERY) with no antibiotic activity, has been shown to bind to motilin receptors and stimulate contractile activity of the antrum and small intestine. The objective of this study was to determine the effect of ABT-229 on canine gastric emptying (GE) and contractile activity of the antrum and duodenum in response to a solid meal. Six beagles were used to determine GE of a solid meal and contractile activity in response to either vehicle, ABT-229 (0.17, 0.83, 2.5, or 5.0 microg/kg/min), ERY (33.3 microg/kg/min), or cisapride (CIS) (10 microg/kg/min). Lag (t(lag)), half-emptying (t(1/2)), and complete emptying (t(full)) times were determined. Contractile data were analyzed for motility index and gastroduodenal coordination. Compared with vehicle, ABT-229 dose dependently accelerated GE, t(lag) was decreased at the two highest doses, t(1/2) was decreased compared with vehicle at the three highest doses, and t(full) was decreased at all doses compared with vehicle. ERY also decreased t(1/2) and t(full), whereas CIS decreased all GE parameters. The slopes of the linear phase of GE curves for all drugs and doses were greater than those for vehicle. ABT-229 dose dependently increased the motility index as well as gastroduodenal coordination. ABT-229 (two highest doses) and CIS accelerated GE of a solid meal by decreasing the lag phase and increasing the rate of GE, whereas ERY only increased the rate of GE. The data suggest that ABT-229 is 7- to 40-fold more potent than ERY in accelerating GE.
Assuntos
Cisaprida/farmacologia , Eritromicina/análogos & derivados , Eritromicina/farmacologia , Esvaziamento Gástrico/efeitos dos fármacos , Fármacos Gastrointestinais/farmacologia , Animais , Cães , Relação Dose-Resposta a Droga , Motilidade Gastrointestinal/efeitos dos fármacos , Contração Muscular/efeitos dos fármacosRESUMO
To test the hypothesis that neutrophil influx is important for the removal of necrotic airway epithelial cells, rhesus monkeys were treated with a function-blocking monoclonal antibody (MAb) against CD18 followed by exposure to ozone or filtered air. CD18 MAb-treated, ozone-exposed monkeys showed a significant inhibition of neutrophil emigration and an accumulation of necrotic airway epithelial cells. In a subsequent experiment, monkeys were given CD18 MAb or an isotype control immunoglobulin before ozone or filtered-air exposure. Complement 5a was instilled into lobes of the right lung at the end of the exposure. Lavage neutrophils were significantly elevated in the right lobes compared with those in the contralateral left lobes; consequently, there were significantly fewer necrotic cells in the airways of the right lung, whereas large aggregations of necrotic cells were observed in the contralateral airways of the left lung. These data indicate that neutrophil influx in ozone-induced injury in primates is CD18 dependent and that neutrophils contribute to the repair of airway epithelium by removal of injured epithelial cells.
Assuntos
Células Epiteliais/patologia , Pneumopatias/patologia , Neutrófilos/imunologia , Oxidantes Fotoquímicos/efeitos adversos , Ozônio/efeitos adversos , Animais , Anticorpos Monoclonais/farmacologia , Bromodesoxiuridina/análise , Brônquios/imunologia , Brônquios/patologia , Líquido da Lavagem Broncoalveolar/imunologia , Antígenos CD18/imunologia , Complemento C5a/imunologia , Células Epiteliais/química , Células Epiteliais/imunologia , Pneumopatias/induzido quimicamente , Macaca mulatta , Masculino , Microscopia Confocal , NecroseRESUMO
Rapamycin, a macrolide antibiotic known to prevent allograft rejection, is a potent inhibitor of cell proliferation. Therefore we studied the effects of orally administered rapamycin in a pig model of balloon injury in an attempt to reduce the cellular proliferation and neointimal formation thought to play a role in restenosis. Twenty Yucatan minipigs, divided into groups of 10 animals each, were subjected to balloon inflation of the carotid arteries. One group received the methylcellulose vehicle for rapamycin, whereas the second group was treated for a total of 31 days with 2.0 mg/kg of rapamycin administered daily by oral gavage. This dose and treatment regimen produced significant (p < 0.05) reductions in neointimal area (59%) and in the maximal thickness of the neointima (59%) when comparisons were made with vehicle-treated animals. These effects were accompanied by a significant increase in the lumen area in animals that received rapamycin (33%). Medial area was decreased by 18% in these animals. Blood samples from rapamycin-treated pigs indicated peak concentrations of 1.87 +/- 0.45 and 1.70 +/- 0.24 ng/ml at 2 and 4 weeks after balloon angioplasty, respectively. Significant increases in blood pressure of 21 mm Hg and decreases in heart rate of 25 beats/min also were observed in rapamycin-treated animals relative to those that received vehicle. These results indicate that the antiproliferative effect of rapamycin can be demonstrated after oral dosing in a pig vascular injury model, suggesting a possible therapeutic utility for rapamycin or its analogs in patients undergoing balloon angioplasty.
Assuntos
Angioplastia com Balão/efeitos adversos , Antibacterianos/uso terapêutico , Estenose das Carótidas/prevenção & controle , Sirolimo/uso terapêutico , Túnica Íntima/metabolismo , Animais , Pressão Sanguínea/efeitos dos fármacos , Artérias Carótidas/patologia , Artérias Carótidas/cirurgia , Lesões das Artérias Carótidas , Divisão Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Frequência Cardíaca/efeitos dos fármacos , Linfócitos/metabolismo , Masculino , Sirolimo/farmacocinética , Suínos , Porco Miniatura , Túnica Íntima/efeitos dos fármacosRESUMO
Catheter-directed thrombolysis has gained increasing acceptance for the treatment of patients who present with vascular occlusion; however, intravenous injection may be preferable in selected patients. Recombinant prourokinase (r-proUK) is a recently-developed fibrin-selective thrombolytic agent with specificity for clot-bound plasminogen. To compare the effects of r-proUK on clot lysis and restoration of blood flow when injected by either intraarterial or intravenous routes of administration, we utilized a dog model of arterial thrombosis in which a radiolabelled clot is formed in the femoral artery. The r-proUK was given by intravenous infusion to one group of 18 animals in doses ranging from 10,000 IU/kg to 100,000 IU/kg; a second group of 27 dogs was treated with r-proUK administered by the intra-arterial route in a dose range from 300 IU to 10,000 IU. Clot lysis was measured by monitoring the loss of counts from the radiolabelled clot over time; blood flow was also monitored throughout the experimental period. Animals which received intravenous treatment showed dose-related clot lysis ranging from 14% to 70% at 2 h, while those which received intra-arterial infusions showed lysis ranging from 22% to 79% over the same period. For similar degrees of clot lysis attained at the highest dose levels of 100,000 IU/kg and 10,000 IU, blood flow was restored to 77% and 35% of control levels in dogs which received intravenous and intraarterial treatment, respectively. The hemostatic protein fibrinogen was not reduced in any of the treatment groups. The results indicate that 100 times more intravenous than intra-arterial r-proUK is required to produce similar clot lysis in this canine model, and that the agent can be administered at this level without induction of a systemic lytic state.
Assuntos
Arteriopatias Oclusivas/tratamento farmacológico , Artéria Femoral , Fibrinolíticos/uso terapêutico , Terapia Trombolítica , Trombose/tratamento farmacológico , Ativador de Plasminogênio Tipo Uroquinase/uso terapêutico , Animais , Cães , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos , Fibrinolíticos/administração & dosagem , Membro Posterior/irrigação sanguínea , Infusões Intra-Arteriais , Infusões Intravenosas , Injeções Intravenosas , Masculino , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/uso terapêutico , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/genética , Proteínas Recombinantes/uso terapêutico , Fluxo Sanguíneo Regional/efeitos dos fármacos , Ativador de Plasminogênio Tipo Uroquinase/administração & dosagem , Ativador de Plasminogênio Tipo Uroquinase/genéticaRESUMO
Platelet-activating factor (PAF) may be an important mediator of allergic rhinitis. In the present study we evaluated the effectiveness of a recently described PAF antagonist (ABT-491) in rat and guinea pig models of allergic rhinitis. PAF, when perfused through the nasal passages of anesthetized Brown Norway rats, provoked an acute increase, measured as dye leakage, in nasal vascular permeability evident within 15 min after exposure to PAF. ABT-491, given orally 1 hr before PAF challenge, inhibited the response in a dose-related manner (ED50 = 0.3 mg/kg). Intranasal perfusion with ovalbumin in rats sensitized to the antigen 18 to 21 days before challenge also induced an increase in vascular permeability. The antigen-induced leakage was inhibited a maximum of 74% (P < or = .001) by pretreatment with ABT-491 (3 mg/kg p.o.). An antihistamine (mepyramine, 10 mg/kg i.p.), a serotonin antagonist (methysergide) and a 5-lipoxygenase inhibitor (A-79175) also exhibited efficacy in this model (56%, 87% and 65% inhibition, respectively). Nearly complete inhibition (93%, P < or = .001) of the response was achieved by coadministration of ABT-491 and methysergide. In guinea pigs intranasal administration of PAF resulted in increased airway resistance that was inhibited in a dose-dependent manner by oral administration of ABT-491 (ED50 = 1 mg/kg). Antigen-induced nasal airway resistance, triggered by exposure of sensitized animals to aerosolized ovalbumin, was also inhibited by ABT-491 (maximum inhibition 64%, P < or = .05, 10 mg/kg p.o.). The effectiveness of the antagonist was increased to 80% protection by coadministration with either an antihistamine or a 5-lipoxygenase inhibitor, agents which were separately insignificant in blocking the response to antigen. These results suggest a therapeutic utility for ABT-491, perhaps in combination with other anti-inflammatory agents, in the treatment of allergic rhinitis.
Assuntos
Hipersensibilidade/tratamento farmacológico , Imidazóis/farmacologia , Indóis/farmacologia , Fator de Ativação de Plaquetas/antagonistas & inibidores , Rinite/tratamento farmacológico , Resistência das Vias Respiratórias/efeitos dos fármacos , Animais , Permeabilidade Capilar/efeitos dos fármacos , Cobaias , Masculino , Fator de Ativação de Plaquetas/fisiologia , Ratos , Ratos Endogâmicos BNRESUMO
To investigate the cellular immune events contributing to airway hyperreactivity (AHR), we studied an in vivo mouse model induced by the hapten picryl (trinitrophenyl) chloride (PCl). Mice were immunized by cutaneous contact sensitization with PCl and airway challenged subsequently with picryl sulfonic acid (PSA) antigen (Ag). Increased airway resistance was produced late (24 h) after Ag challenge, disappeared by 48 h, and was associated with no decrease in diffusion capacity. AHR could be produced in PCl immune/ PSA challenged mice on day 7 or even, with challenge, as early as 1 d after contact sensitization, after adoptive transfer of immune cells lacking CD3(+) contact sensitivity effector T cells, or after transfer of Ag-specific lymphoid cells depleted of conventional T lymphocytes with surface determinants for CD3, CD4, CD8, TCR-beta, or TCR-delta molecules. Further experiments showed that development of AHR depended upon transfer of immune cells expressing surface membrane Thy-1 and B220 (CD45RA) determinants. We concluded that a novel population of Ag-specific lymphoid cells with a defined surface phenotype (Thy-1(+), CD3(-), CD4(-), CD8(-), TCR-alphabeta-, TCR-gammadelta-, and CD45RA+) is required in a mouse model for the development of AHR.
Assuntos
Transferência Adotiva , Asma/imunologia , Complexo CD3/imunologia , Imunidade Celular , Antígenos Comuns de Leucócito/imunologia , Linfócitos T/imunologia , Antígenos Thy-1/imunologia , Animais , Asma/fisiopatologia , Feminino , Haptenos/administração & dosagem , Haptenos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Cloreto de Picrila/administração & dosagem , Cloreto de Picrila/imunologiaRESUMO
Balloon angioplasty has become an important intervention in clinical cardiology; however, the technique is associated with a high incidence of restenosis, requiring repeated procedures. Endothelin-1 (ET-1), specifically through its action on ET(A) receptors, has been implicated in the cell proliferation and subsequent neointimal formation that leads to restenosis. Therefore we examined a potent antagonist of the ET(A) receptor, A127722.5, in a pig model of balloon angioplasty in iliac and carotid arteries. Ten pigs received A-127722.5 (7.5 mg/kg b.i.d.) orally, starting 3 days before angioplasty and continuing for 4 weeks; 10 additional pigs were treated with the same dosing regimen of the angiotensin-converting enzyme (ACE) inhibitor captopril (3.0 mg/kg b.i.d.), while a third group of 10 animals received placebo. At 2 and 4 weeks after the start of treatment, these doses of the ET(A) receptor antagonist and ACE inhibitor blocked the presser responses induced by big ET-1 and angiotensin I, respectively. In the iliac arteries, neointimal formation, neointimal/medial ratio, and maximal neointimal thickness were all significantly reduced, and the residual lumen area was significantly increased in pigs treated with the ET(A) receptor antagonist compared with placebo and captopril-treated groups. Medial collagen content, collagen deposition, and medial growth also were significantly reduced relative to the placebo group. Beneficial effects also were observed in the carotid arteries, although the results were less striking. Captopril was ineffective in protecting against the effects of balloon angioplasty in both vessels. Our results indicate that an orally active and potent antagonist of the ET(A) receptor inhibits cell proliferation and synthesis of extracellular matrix in pigs and may provide an important therapeutic approach to the prevention of restenosis.
Assuntos
Angioplastia com Balão/efeitos adversos , Antagonistas dos Receptores de Endotelina , Músculo Liso Vascular/efeitos dos fármacos , Pirrolidinas/farmacologia , Angiotensina I/farmacologia , Inibidores da Enzima Conversora de Angiotensina/farmacologia , Animais , Atrasentana , Pressão Sanguínea/efeitos dos fármacos , Captopril/farmacologia , Lesões das Artérias Carótidas , Artéria Carótida Primitiva/efeitos dos fármacos , Artéria Carótida Primitiva/metabolismo , Artéria Carótida Primitiva/patologia , Colágeno/biossíntese , Endotelina-1/sangue , Endotelina-1/farmacologia , Hiperplasia , Artéria Ilíaca/efeitos dos fármacos , Artéria Ilíaca/lesões , Artéria Ilíaca/metabolismo , Artéria Ilíaca/patologia , Masculino , Músculo Liso Vascular/lesões , Músculo Liso Vascular/metabolismo , Músculo Liso Vascular/patologia , Pirrolidinas/sangue , Receptor de Endotelina A , Suínos , Porco Miniatura , Túnica Íntima/efeitos dos fármacos , Túnica Íntima/patologiaRESUMO
Endothelin has been implicated in the pathogenesis of acute renal failure associated with radiocontrast media. The current study was designed to determine the effect of endothelin receptor blockade in a model of radiocontrast-induced nephropathy. Inhibition of prostanoids and nitric oxide with indomethacin and L-nitroarginine methyl ester (L-NAME) predisposes rats to the nephrotoxic effects of radiocontrast media and, therefore, has been proposed as a clinically relevant model. Separate groups of conscious rats were given the ETA receptor antagonist, A-127722 (3, 10, or 30 mg/kg), or water (1 mL/kg) by oral gavage. All rats were then given indomethacin (10 mg/kg), L-NAME (10 mg/kg), and the contrast agent, diatrizoate (6 mL/kg), by intravenous injection at 15-min intervals. Urine was collected for the subsequent 24 h by placing rats in metabolism cages. When indomethacin, L-NAME, and diatrizoate were administered without A-127722, rats displayed typical signs of renal failure; protein excretion was increased from a baseline of 13 +/- 2 to 33 +/- 8 mg/day (p < 0.05) and plasma creatinine increased from 0.52 +/- 0.01 to 0.62 +/- 0.04 mg/dL (p < 0.05). ETA receptor blockade prevented the rise in protein excretion and plasma creatinine in a dose-dependent manner. In separate series of clearance experiments, A-127722 completely inhibited the renal effects of exogenous ET-1. These results suggest that endothelin plays a role in the hemodynamic events in this model and that ETA receptor antagonists should be investigated as potential therapeutic agents for radiocontrast-induced nephropathy.
Assuntos
Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/prevenção & controle , Meios de Contraste/efeitos adversos , Diatrizoato/efeitos adversos , Antagonistas dos Receptores de Endotelina , Pirrolidinas/uso terapêutico , Animais , Atrasentana , Creatinina/sangue , Modelos Animais de Doenças , Endotelinas/fisiologia , Indometacina/farmacologia , Masculino , NG-Nitroarginina Metil Éster/farmacologia , Óxido Nítrico/antagonistas & inibidores , Antagonistas de Prostaglandina/farmacologia , Proteinúria/urina , Ratos , Ratos Sprague-Dawley , EstereoisomerismoRESUMO
Using various animal models of toxic or antigenic-induced airway inflammation, we have demonstrated that adhesion molecules play an important role in the recruitment, retention, and site-specific activation of inflammatory cells within the airways. Furthermore, we have shown that cytokines may contribute to inflammatory responses in the airways by enhancing the expression of adhesion molecules on respiratory epithelial cells.
Assuntos
Bronquite/metabolismo , Moléculas de Adesão Celular/metabolismo , Pneumonia/metabolismo , Alvéolos Pulmonares/metabolismo , Animais , Brônquios/metabolismo , Linhagem Celular , Células Cultivadas/metabolismo , Doença Crônica , Modelos Animais de Doenças , Epitélio/metabolismo , Imuno-Histoquímica , Molécula 1 de Adesão Intercelular , Macaca fascicularis , Masculino , Camundongos , Oxigênio/toxicidadeRESUMO
The distribution of intercellular adhesion molecule-1 (ICAM-1) on alveolar epithelial cells and the effects of exposure to 100% O2 on ICAM-1 expression in mouse lungs were studied by EM immunocytochemistry and immunoblot analysis. Cryoultrathin sections from mouse lungs exposed to air or 100% O2 for 84 h were labeled with a monoclonal rat anti-mouse ICAM-1 antibody. In the normal lung, abundant ICAM-1 expression was found on the alveolar surface of type I epithelial cells. Furthermore, ICAM-1 is highly concentrated on the surfaces near cell junctions. ICAM-1 was also found on the capillary surface of endothelial cells and alveolar surface of type II cells at densities considerably lower than that found on type I epithelial cells. After exposure to O2, the labeling density of ICAM-1 on the central surface of type I epithelial cells was not changed significantly. However, the gradient of ICAM-1 on the surfaces near cell junctions was nearly abolished. ICAM-1 labeling on the capillary surface of endothelial cells remained low. ICAM-1 was also markedly induced on the alveolar surface of type II epithelial cells after hyperoxic exposure. These results show that ICAM-1 is expressed primarily on type I epithelial cell surfaces near cell junctions. Exposure to hyperoxia causes a dramatic change in the distribution pattern of ICAM-1 on alveolar type I epithelial cells and induces expression of ICAM-1 on alveolar type II epithelial cells. These hyperoxia-induced changes may influence the associated neutrophil invasion/retention in the alveolar air spaces or alveolar walls.
Assuntos
Moléculas de Adesão Celular/biossíntese , Oxigênio/farmacologia , Alvéolos Pulmonares/metabolismo , Análise de Variância , Animais , Epitélio/metabolismo , Molécula 1 de Adesão Intercelular , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia ImunoeletrônicaRESUMO
BI-L-226, a 2,6-disubstituted 4-(2-arylethenyl)phenol, is a potent and selective 5-lipoxygenase inhibitor which shows excellent inhibition of antigen-induced leukotriene generation in the lung of cynomolgus monkeys by aerosol administration, although little activity has been observed by the p.o. route. The facile synthesis of the succinate ester BI-L-357, however, results in a prodrug which has p.o. activity between 10 to 30 mg/kg in an ex vivo whole blood model of leukotriene B4 generation in both squirrel and cynomolgus monkeys. In addition, the prodrug is effective in inhibiting pulmonary leukotriene C4 production in antigen-challenged cynomolgus monkeys in the same dose range. Plasma levels of the parent compound in the monkey after p.o. administration of 30 mg/kg are 25-fold higher than the IC50 needed for in vitro inhibition of leukotriene B4 in whole blood. Absolute bioavailability of the parent compound was 50%. The prodrug concept therefore extends the potential of this class of compounds to inflammation sites mediated by 5-lipoxygenase not readily treated by topical administration.
Assuntos
Inibidores de Lipoxigenase/farmacologia , Fenóis/farmacologia , Pró-Fármacos/farmacologia , Tiofenos/farmacologia , Animais , Antígenos , Disponibilidade Biológica , Calcimicina/farmacologia , Feminino , Humanos , Leucotrieno B4/biossíntese , Leucotrieno B4/sangue , Inibidores de Lipoxigenase/farmacocinética , Pulmão/metabolismo , Macaca fascicularis , Masculino , Fenóis/sangue , Fenóis/farmacocinética , Pró-Fármacos/farmacocinética , SRS-A/antagonistas & inibidores , SRS-A/biossíntese , Saimiri , Tiofenos/sangue , Tiofenos/farmacocinética , Tromboxano B2/biossíntese , Tromboxano B2/sangueRESUMO
BACKGROUND: The action of 5-lipoxygenase on arachidonic acid generates potent inflammatory mediators that may contribute to the pathophysiology of asthma. METHODS: Using the potent and selective 5-lipoxygenase inhibitor BI-L-239, we have examined the role of 5-lipoxygenase products in three animal models of asthma. RESULTS: In vitro BI-L-239 inhibited 5-lipoxygenase product generation from human lung mast cells, alveolar macrophages, and peripheral blood leukocytes with a concentration that would provide 50% inhibition values of 28 to 340 nmol/L. A 36-fold selectivity for immunoreactive leukotriene C4 versus immunoreactive prostaglandin D2 inhibition was demonstrated in mast cells. In anesthetized cynomolgus monkeys, inhaled BI-L-239 provided dose-dependent inhibition of the inhaled Ascaris-induced immunoreactive leukotriene C4 release (maximum, 73%; bronchoalveolar lavage [BAL], 20 minutes), late-phase bronchoconstriction (maximum, 41%; +6 to 8 hours), and neutrophil infiltration (maximum, 63%; BAL, +8 hours). In conscious sheep, inhaled BI-L-239 provided dose-dependent inhibition of the inhaled Ascaris-induced late-phase bronchoconstriction (maximum, 66%; +6 to 8 hours) and increase in airway responsiveness (maximum, 82%; carbachol, +24 hours). The acute bronchoconstriction was shortened, and neutrophil infiltration diminished (maximum, 61%; BAL, +8 hours) in this model. Finally in conscious actively sensitized guinea pigs pretreated with pyrilamine and indomethacin, inhaled BI-L-239 attenuated acute bronchoconstriction (maximum, 80%; +5 to 15 minutes), leukocyte infiltration (58%; BAL, +3 days) and increase in airway responsiveness (100%; methacholine, +3 days) induced by three alternate-day ovalbumin inhalations. CONCLUSIONS: In conclusion, results in these three animal models indicate that 5-lipoxygenase products may be major contributors to the bronchoconstriction (especially late phase), leukocyte infiltration, and airway hyperresponsiveness that characterize asthma.
Assuntos
Araquidonato 5-Lipoxigenase/fisiologia , Asma/fisiopatologia , Broncoconstrição , Inibidores de Lipoxigenase/farmacologia , Fenóis/farmacologia , Animais , Relação Dose-Resposta a Droga , Cobaias , Humanos , Macaca fascicularis , Masculino , Prostaglandina D2/metabolismo , SRS-A/metabolismo , OvinosAssuntos
Antígenos/administração & dosagem , Ativação Linfocitária/imunologia , Macaca fascicularis/imunologia , Subpopulações de Linfócitos T/imunologia , Animais , Hiper-Reatividade Brônquica/imunologia , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Masculino , Fatores de TempoRESUMO
Cell surface adhesive glycoproteins are principal regulators of nearly all aspects of immune/inflammatory responses. Using monoclonal antibodies to individual adhesion molecules, the expression and contribution of specific molecules in the pathogenesis of allergen-induced airway hyperresponsiveness in monkeys has been studied. Results confirm the importance of cell adhesion and demonstrate that antagonism of a single adhesion molecule may provide a novel therapeutic approach.