RESUMO
Anti-islet immunity was studied in six spontaneously insulin-dependent diabetic (IDD) dogs, using mouse islets of Langerhans cells as targets, in vitro. Insulinopenia was demonstrated in all dogs by an i.v. glucose tolerance test. A significant lymphocytopenia was detected in the peripheral blood of this diabetic group. Pancreatic tissue from one of these animals was obtained shortly after death and the islets displayed a marked loss in beta cells without significant changes in the other types of islet cells. No insulitis was observed. Circulating mononuclear cells from the diabetic dogs induced an increased basal insulin (IRI) release from islet cells and a suppressed stimulated IRI release. Damage to or depth of beta cells may account for these findings. The stimulated IRI release was also suppressed when islets were incubated with the diabetic sera + complement, while the D-cell response to arginine was not altered, and the A-cell response was reduced but not abolished. A lysis of islet cells in the presence of IDD sera + complement was demonstrated by an increased release of 51Cr from labeled cells. These anomalies were observed neither when complement was heat-inactivated nor in the presence of control sera + complement. Canine IDD may be a new animal model for the study of anti-islet cellular and humoral immunities.
Assuntos
Diabetes Mellitus Tipo 1/imunologia , Ilhotas Pancreáticas/imunologia , Animais , Cromo/metabolismo , Cães , Teste de Tolerância a Glucose , Humanos , Insulina/sangue , Contagem de Leucócitos , Linfócitos/imunologia , Camundongos , RatosRESUMO
Isolated mouse vas deferens preparations were used to study the effect of temperature on noradrenaline-induced contractions. Preparations were suspended in the organ bath containing Krebs-Henseleit solution for isometric tension recording. Contractile responses to noradrenaline were investigated in the mouse vas deferens after moderate cooling from 37 to 26 or 22 degrees C. A significant increase of the phasic contractions to noradrenaline was observed at 26 or 22 degrees C compared with responses obtained at 37 degrees C (about 12.3 and 35.6% increase at 26 and 22 degrees C, respectively). The secondary noradrenaline-induced sustained contraction was also significantly enhanced after moderate cooling to 26 degrees C. The potentiation of noradrenaline-induced contraction at 26 degrees C remained in a Ca(2+)-free EGTA (1 mM)-containing solution. However, sustained contraction was suppressed after removal of the calcium from the medium at 37 and 26 degrees C. Contraction to caffeine was significantly enhanced at 22 degrees C compared with 37 degrees C. By contrast, barium chloride-induced contraction of the vas deferens was markedly decreased after moderate cooling to 22 degrees C. In the presence of ouabain (0.1 mM), the noradrenaline-induced peak contraction was significantly increased at 37 degrees C. However, potentiation of the noradrenaline response at 22 degrees C was unaffected by the Na+/K+ pump inhibitor. Noradrenaline-induced peak contractions were depressed in the presence of vanadate (1 mM) and cyclopiazonic acid (10 microM), two Ca(2+)-ATPase inhibitors, at 37 degrees C and also at 22 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Cálcio/metabolismo , Temperatura Baixa , Inibidores Enzimáticos/farmacologia , Músculo Liso/fisiologia , Norepinefrina/farmacologia , Ducto Deferente/fisiologia , Animais , Compostos de Bário/farmacologia , Cafeína/farmacologia , ATPases Transportadoras de Cálcio/antagonistas & inibidores , Cloretos/farmacologia , Indóis/farmacologia , Contração Isométrica/efeitos dos fármacos , Masculino , Camundongos , Músculo Liso/efeitos dos fármacos , Ouabaína/farmacologia , ATPase Trocadora de Sódio-Potássio/metabolismo , Vanadatos/farmacologia , Ducto Deferente/efeitos dos fármacos , Ducto Deferente/metabolismoRESUMO
In the present study we have investigated the involvement of sensitized mice immunoglobulins and some electrophysiological alterations that participate to the antigenic sensitization-induced hyperreactivity of isolated mouse vas deferens. Active sensitization was performed by subcutaneous injection of egg albumen. Contractile responses to noradrenaline were isometrically recorded in the isolated vas deferens. Low external Na(+)-induced contractions and rapid cooling contractures were evaluated. Resting membrane potential (Er) and intracellular Na activity were measured in control and actively sensitized vas deferens by using conventional KCl-filled and Na(+)-sensitive microelectrodes respectively. Active sensitization-induced hyperreactivity to noradrenaline was reproduced by in vitro passive sensitization of control vas deferens with sensitized mice immunoglobulins. The inhibition of the nitric oxide synthesis by N-nitro-L-arginine methyl ester (L-NAME) did not change control vas deferens reactivity in vitro to noradrenaline and acetylcholine. Rapid cooling contractures, performed after lowering external Na+ concentration, were not altered by active sensitization. However, sensitization increased significantly the strength of the low external Na+-induced contractions. In control vas deferens Er was a mean of -49.2+/-0.3 mV (mean+/-SEM). Sensitization resulted in reduction of Er by 14 mV. In sensitized preparations, relative insensitivity of Er to ouabain, external K+ removal and cooling were observed. The intracellular Na+ activity was increased by about 40% in sensitized vas deferens. It is concluded that sensitization-induced hyperreactivity is mediated by immunoglobulins and produced smooth muscle cells depolarisation. The low Er of sensitized muscle may be partly the result of an increase in membrane permeability to Na+ which could interfere with intracellular Ca2+ homeostasis.
Assuntos
Imunoglobulinas/imunologia , Contração Muscular/efeitos dos fármacos , Sódio/metabolismo , Ducto Deferente/fisiologia , Animais , Cálcio/metabolismo , Homeostase , Imunização , Masculino , Potenciais da Membrana , Camundongos , NG-Nitroarginina Metil Éster/farmacologia , Norepinefrina/farmacologia , ATPase Trocadora de Sódio-Potássio/fisiologia , Ducto Deferente/efeitos dos fármacosAssuntos
Animais Domésticos , Anti-Infecciosos/efeitos adversos , Doença Hepática Induzida por Substâncias e Drogas/veterinária , Tratamento Farmacológico/veterinária , Tolerância a Medicamentos , Medicina Veterinária , Anestésicos/efeitos adversos , Animais , Doenças do Gato/tratamento farmacológico , Gatos , Doenças do Cão/tratamento farmacológico , Cães , Fígado/metabolismoAssuntos
Antitussígenos/farmacologia , Prometazina , Animais , Estimulação Elétrica , Cobaias , MasculinoAssuntos
Depressores do Apetite , Morfolinas/farmacologia , Noscapina/farmacologia , Animais , Antitussígenos , Depressores do Apetite/administração & dosagem , Preparações de Ação Retardada , Cães , Cobaias , Camundongos , Morfolinas/administração & dosagem , Noscapina/administração & dosagem , Ratos , Fatores de TempoRESUMO
Changes in the reactivity of the ileum (to histamine and barium chloride) and vas deferens (to acetylcholine and barium chloride), isolated from actively egg albumen-sensitized guinea pigs, have been investigated. The study was performed on 2 guinea pig strains: the Dunkin-Hartley strain, usually used as an airway allergic model, and the BFA strain. In actively sensitized guinea pigs of both strains, concentration-response curves exhibited a significant dose-dependent upward shift compared to those obtained in control guinea pigs. The maximal contraction strength calculated from these curves was significantly enhanced in both sensitized guinea pig strains, without a change in EC50 values. This study showed that the active antigen sensitization procedure involved several smooth muscle functions, and not exclusively the trachea.
Assuntos
Músculo Liso/efeitos dos fármacos , Músculo Liso/imunologia , Acetilcolina/fisiologia , Animais , Compostos de Bário/farmacologia , Cloretos/farmacologia , Cobaias , Histamina/fisiologia , Íleo/efeitos dos fármacos , Íleo/imunologia , Imunização , Técnicas In Vitro , Masculino , Contração Muscular/efeitos dos fármacos , Contração Muscular/imunologia , Ducto Deferente/efeitos dos fármacos , Ducto Deferente/imunologiaRESUMO
1. Dose-response curves for noradrenaline, phenylephrine and clonidine were determined isometrically on the mouse vas deferens at 26 degrees C, 15 degrees C and compared to the one obtained at 37 degrees C. 2. In the presence of noradrenaline, reducing temperature induced an increase of both maximal developed tension and sensitivity to the drug. Reduction by 50% of the extracellular calcium concentration abolished the maximal contraction potentiation. 3. When reducing temperature to 26 degrees C, the maximal contraction was increased and depressed in the presence of phenylephrine and clonidine respectively. 4. The results suggest (a) that cooling increases the reactivity of mouse vas deferens by activation of alpha 1 adrenoceptors and depresses it by activation of alpha 2 adrenoceptors (b) that calcium ions could play an important role in the potentiation of the maximal contraction.
Assuntos
Temperatura Baixa , Simpatomiméticos/farmacologia , Ducto Deferente/efeitos dos fármacos , Animais , Cálcio/fisiologia , Clonidina/farmacologia , Relação Dose-Resposta a Droga , Espaço Extracelular/metabolismo , Técnicas In Vitro , Isoproterenol/farmacologia , Masculino , Camundongos , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Músculo Liso/fisiologia , Norepinefrina/farmacologia , Fenilefrina/farmacologia , Ducto Deferente/fisiologiaRESUMO
BACKGROUND: Vascular hyperresponsiveness can be reproduced by in vitro passive sensitization of isolated aorta with immunoglobulin G1 (IgG1) taken from ovalbumen-sensitized BFA guinea-pig. OBJECTIVE: The aim of the present work was to investigate the role of nitric oxide in the sensitization-induced alteration of the contractile and relaxant responses of guinea-pig aorta to noradrenaline (NA) and acetylcholine (ACh), respectively. METHODS: Cumulative concentration-response curves to NA or ACh were established before and after IgG1 sensitization and then after successive treatments. RESULTS: IgG1 in vitro passive sensitization of aorta caused a significant hyperreactivity to NA and completely inhibited the relaxation to ACh. After sensitization, the addition of an intact aortic ring (with endothelium) in the organ chamber restored the maximal response to NA and ACh close to control but was ineffective in the presence of hemoglobin. The restoration of the control reactivity to NA was also inhibited in the presence of L-NAME or when the added aortic ring was endothelium-denuded. Moreover, L-arginine, a nitric oxide (NO) precursor, was able to restore the control reactivity to NA. CONCLUSION: The present results show that IgG1 in vitro sensitization induced a loss of NO release from the vascular endothelium. This loss of NO probably plays a great role in vascular hyperreactivity by increasing the contractile response and decreasing the relaxant response to mediators and would be a component of allergic diseases pathogenesis.
Assuntos
Endotélio Vascular/metabolismo , Imunoglobulina G/farmacologia , Músculo Liso Vascular/metabolismo , Óxido Nítrico/metabolismo , Acetilcolina/farmacologia , Animais , Aorta/efeitos dos fármacos , Aorta/metabolismo , Endotélio Vascular/efeitos dos fármacos , Cobaias , Técnicas In Vitro , Masculino , Músculo Liso Vascular/efeitos dos fármacos , Norepinefrina/farmacologiaRESUMO
1. The present study was designed to investigate the effect of active sensitization on the responsiveness of mouse vas deferens before and after moderate cooling. Contractile responses to noradrenaline (NA) were isometrically recorded in the vas deferens of control and ovalbumen-sensitized mice at 37 degrees C and 22 degrees C. 2. Enhancement of the vas deferens reactivity to NA was observed in the sensitized vs control mice at 37 degrees C and 22 degrees C (P < 0.01). In sensitized mice, maximal contraction was significantly increased compared with controls, and sensitization-induced hyperresponsiveness was greater at 22 degrees C compared with 37 degrees C. At 37 degrees C, contractile responses to barium chloride were significantly enhanced in the sensitized mice compared with controls, whereas the reduction of the temperature to 22 degrees C produced a marked inhibition of vas deferens contractions in both groups. Caffeine-induced contractions of the vas deferens were similar in control and sensitized mice at 37 degrees C. After moderate cooling to 22 degrees C, vas deferens from sensitized mice became hyperresponsive compared with controls. 3. Ouabain (0.1 mM) produced an increase of NA-induced contraction in control and sensitized vas deferens at 37 degrees C (P < 0.01). It had no significant effect in the control at 22 degrees C but produced a marked inhibition of NA-induced contraction in the sensitized vas deferens at 22 degrees C. Contractions to NA in the presence of vanadate (1 mM) were depressed in control and sensitized mice at both temperatures. 4. These results suggest that sensitization-induced hyperresponsiveness of the mouse vas deferens is mediated by an increased mobilization of intracellular calcium. The involvement of an unknown ouabain-sensitive pathway in sensitization-induced alterations is also discussed.
Assuntos
Músculo Liso/efeitos dos fármacos , Ovalbumina/administração & dosagem , Ducto Deferente/efeitos dos fármacos , Análise de Variância , Animais , Compostos de Bário/farmacologia , Cafeína/farmacologia , Cálcio/metabolismo , Estimulantes do Sistema Nervoso Central/farmacologia , Cloretos/farmacologia , Temperatura Baixa , Inibidores Enzimáticos/farmacologia , Contração Isométrica/efeitos dos fármacos , Masculino , Camundongos , Músculo Liso/fisiologia , Norepinefrina/farmacologia , Ouabaína/farmacologia , Distribuição Aleatória , Ducto Deferente/fisiologiaRESUMO
BACKGROUND: Smooth muscles hyperresponsiveness is a common feature in anaphylaxis and allergic diseases. OBJECTIVE: The aim of the present work was to investigate the effect of in vitro passive sensitization with highly purified immunoglobulin G1 (IgG1) on the responsiveness of tracheal, aortic, vas deferens and ileum smooth muscles. METHODS: Firstly, IgG1, obtained from actively sensitized BFA guinea-pigs, was purified by Protein A-Sepharose column and characterized by enzyme-linked immunosorbent assay (ELISA) and immunoelectrophoresis analysis. Concentration-response curves to spasmogens (acetylcholine for trachea and vas deferens, noradrenaline for aorta and histamine for ileum) were established before and after in vitro passive sensitization with IgG1. RESULTS: Contractile responses and maximal contractions were significantly enhanced after passive sensitization for all the organs. Maximal contractions were significantly increased in the trachea (+46.7%), aorta (+51%), vas deferens (+114.2%) and ileum (+117.2%). At the end of the experiments, the application of the sensitizing antigen induced a significant Schultz-Dale reaction of the smooth muscles. CONCLUSION: The present results show that the in vitro application of purified IgG1 can produce non-specific smooth muscle hyperreactivity and hypersensitivity. So, IgG1 can be considered as the main factor involved in the genesis of sensitization-induced hyperresponsiveness, and probably play a great role in hyperreactivity observed during allergic diseases and anaphylaxis.
Assuntos
Imunoglobulina G/farmacologia , Contração Muscular/imunologia , Músculo Liso/imunologia , Animais , Antígenos/imunologia , Aorta/efeitos dos fármacos , Aorta/imunologia , Cromatografia de Afinidade , Eletroforese em Gel de Poliacrilamida , Cobaias , Íleo/efeitos dos fármacos , Íleo/imunologia , Imunização , Imunoglobulina G/isolamento & purificação , Masculino , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Traqueia/efeitos dos fármacos , Traqueia/imunologia , Ducto Deferente/efeitos dos fármacos , Ducto Deferente/imunologiaRESUMO
The purpose of this study was to evaluate the strain-related differences in tracheal hyperresponsiveness in control and egg albumen-sensitized guinea pigs. Concentration-response curves to acetylcholine and barium chloride were established from tracheal rings of Dunkin-Hartley and BFA strain guinea pigs. In the Dunkin-Hartley strain, sensitization did not significantly increase the tracheal responsiveness to acetylcholine and barium chloride. By contrast, in the BFA strain, significant sensitization-induced hyperreactivity was achieved as the maximal contractions induced by acetylcholine and barium chloride, were enhanced from 6.5 +/- 1.2 and 3.2 +/- 0.4 mN in control to 10.0 +/- 1.4 and 5.6 +/- 0.8 mN, respectively, in sensitized animals. However, antigen challenge, performed in vitro, exhibited a similar amplitude of contraction in tracheal rings from both strains (Dunkin-Hartley 5.1 +/- 0.8 mN; BFA 5.9 +/- 0.5 mN). Finally, while the two guinea-pig strains developed specific sensitization to allergen, only tracheal rings from the BFA strain developed hyperresponsiveness to acetylcholine and barium chloride. The strain-related difference appears to be partly explained by a lower basal reactivity in the BFA strain both acetylcholine (Em 7.3 +/- 1.7 and 6.5 +/- 1.2 mN for Dunkin-Hartley and BFA, respectively) and barium chloride (Em 9.4 +/- 2.6 and 3.2 +/- 0.4 mN for Dunkin-Hartley and BFA, respectively). As the same procedure of sensitization provides different results in the genesis of hyperreactivity between the two guinea-pig strains used for asthma models, the BFA guinea-pig strain seems to be a better model because sensitized non-challenged animals could easily be dissociated from control ones, similar to that which occurs in asthmatic patients during provocation tests with cholinergic drugs.
Assuntos
Imunização , Traqueia/fisiopatologia , Acetilcolina/farmacologia , Albuminas/administração & dosagem , Animais , Compostos de Bário/farmacologia , Cloretos/farmacologia , Relação Dose-Resposta a Droga , Cobaias , Masculino , Contração Muscular/efeitos dos fármacos , Contração Muscular/imunologia , Músculo Liso/efeitos dos fármacos , Especificidade da Espécie , Traqueia/efeitos dos fármacos , Traqueia/imunologiaRESUMO
Smooth muscles hyperresponsiveness is a common feature in anaphylaxis and allergic diseases. The aim of the present work was to investigate whether the enhanced reactivity of sensitized guinea-pig vas deferens was associated with changes in the resting membrane potential (Er) of the smooth muscle cells. Active sensitization was performed by subcutaneous injection of egg albumen. Er was measured in vitro in isolated vas deferens with conventional KCl-filled microelectrodes. Quantification of [3H]ouabain binding sites, measurements of 86Rb efflux, and measurements of Na and K contents were also performed. In normal physiological solution, at 35 degrees C, Er was a mean of -54.1+/-0.3 mV (mean +/- SEM) in control vas deferens. Sensitization resulted in depolarizing Er by about 7 mV. In control and sensitized preparations, the 3H-ouabain binding site concentration, the efflux of 86Rb, and the K content were similar. In guinea-pig vas deferens, active sensitization induced a partial depolarization of the resting membrane potential of the smooth muscle cells, which did not result from a downregulation of Na+ -K+ pump sites.