RESUMO
BACKGROUND: Adrenaline is known to inhibit the immunological activation of human basophils. The same effect can be obtained with ultra-low doses of histamine. MATERIAL/METHODS: The present investigation was undertaken to verify the effects of ultra-low doses of adrenaline on the immunological activation of human basophils. For this purpose, purified human basophils were isolated from the blood of atopic donors and the response of the basophils to anti-IgE antiserum was evaluated by measuring the release of histamine and the expressions of the activation markers CD63 and CD203c on the basophil surface. RESULTS: The antigenic activation of basophils was significantly decreased by adrenaline at both pharmacological and ultra-low doses. The inhibiting effects of adrenaline were reversed by the beta blocker propranolol at either dose level. CONCLUSIONS: Although seemingly unexplainable by the currently accepted pharmacological principles, this phenomenon, already reported for histamine, deserves further investigation.
Assuntos
Basófilos/efeitos dos fármacos , Basófilos/imunologia , Epinefrina/administração & dosagem , Epinefrina/farmacologia , Anticorpos Anti-Idiotípicos/imunologia , Antígenos/imunologia , Antígenos CD/metabolismo , Basófilos/citologia , Relação Dose-Resposta a Droga , Citometria de Fluxo , Liberação de Histamina/efeitos dos fármacos , Humanos , Diester Fosfórico Hidrolases/metabolismo , Glicoproteínas da Membrana de Plaquetas/metabolismo , Propranolol/farmacologia , Pirofosfatases/metabolismo , Tetraspanina 30RESUMO
We hypothesized that circulating polymorphonuclear granulocytes (PMNs), vascular endothelial cells (ECs), and perivascular mast cells (MCs) may initiate and sustain the inflammatory response through the generation of the superoxide anion (O(2)(*-)) by PMNs primed by inflammatory stimuli, which in turn evoked the overexpression of adhesion molecules from ECs and release of histamine by MCs. To pin-point the role of carbon monoxide (CO) in curbing vascular inflammation, we studied the effect of a water-soluble CO-releasing molecule [tricarbonylchloro-glycinate-ruthenium (II); CORM-3] on an experimental model of vascular inflammation. The model consists of coincubating formyl-methionyl peptide (fMLP) -primed human PMNs with rat ECs or with rat MCs. The effects of CORM-3 were evaluated by measuring the generation of O(2)(*-) and the expression of CD11b in fMLP-primed PMNs; the expression of ICAM-1 and CD203c in ECs and MCs, respectively; and the release of histamine from MCs. Our results show that the chemotactic peptide fMLP primes PMNs to generate O(2)(*-) and overexpress CD11b, both events being central to the inflammatory process, while CORM-3 significantly decreases these events (IC(50)=1.66 microM for O(2)(*-) production; 1.20 microM for CD11b expression in human PMNs). The experiments also show that fMLP-primed PMNs increase the CD54 expression by coincubated ECs, and the expression of CD203c and the release of histamine by coincubated MCs. Once again, CORM-3 abolishes these events (IC(50)=6.78 microM for CD54 expression in ECs; 1.18 microM for CD203 expression; 1.15 microM for histamine release in MCs). Thus, CORM-3 exerts a powerful anti-inflammatory action by down-regulating the oxidative burst in PMNs, the overexpression of adhesion molecules in PMNs and ECs, the release of histamine, and the overexpression of an activation marker by MCs.
Assuntos
Células Endoteliais/fisiologia , Mastócitos/fisiologia , Neutrófilos/fisiologia , Compostos Organometálicos/farmacologia , Animais , Sobrevivência Celular , Endotélio Vascular/citologia , Masculino , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Ratos , Superóxido Dismutase/metabolismo , Superóxidos/metabolismoRESUMO
Splanchnic artery occlusion (SAO) followed by reperfusion causes endothelial injury and inflammation which contribute to the pathophysiology of shock. We investigated the effects of pea seedling (Latyrus cicera) histaminase, known to afford protection against the deleterious effects of cardiac ischemia/reperfusion, given to rats subjected to SAO/reperfusion-induced splanchnic injury. Histaminase (80 IU kg, 15 min before reperfusion) significantly reduced the drop of blood pressure and high mortality rate caused by SAO/reperfusion. Histaminase also reduced histopathological changes, leukocyte infiltration (myeloperoxidase), and expression of endothelial cell adhesion molecules in the ileum. Histaminase counteracted free radical-mediated tissue injury, as judged by a significant decrease in the plasma and tissue levels of peroxidation and nitration products (oxidized rhodamine, malondialdehyde, nitrotyrosine), DNA damage markers (8-hydroxy-2'-deoxyguanosine, poly-adenosine diphosphate-ribosylated DNA) and consumption of tissue antioxidant enzymes (superoxide dismutase). As a result, histaminase led to a reduction of ileal cell apoptosis (caspase 3, terminal deoxynucleotidyltransferase-mediated UTP end labeling-positive cells). These results show that histaminase exerts a clear-cut protective effect in SAO/reperfusion-induced splanchnic injury, likely caused by oxidative catabolism of proinflammatory histamine and antioxidant effects resulting in hindrance of free radical-mediated tissue injury, endothelial dysfunction, and leukocyte recruitment. Thus, histaminase could be used therapeutically in intestinal ischemia.
Assuntos
Amina Oxidase (contendo Cobre)/uso terapêutico , Arteriopatias Oclusivas/tratamento farmacológico , Proteínas de Plantas/uso terapêutico , Traumatismo por Reperfusão/tratamento farmacológico , Circulação Esplâncnica/efeitos dos fármacos , Animais , Modelos Animais de Doenças , Enteropatias/tratamento farmacológico , Isquemia/tratamento farmacológico , Pisum sativum/enzimologia , Ratos , Ratos WistarRESUMO
INTRODUCTION: Fatalities due to mushroom poisonings are increasing worldwide, with more than 90% of deaths resulting from ingestion of amatoxin-containing species. METHODS: A retrospective evaluation of the history and clinical outcome of each patient treated from 1988 to 2002 in the Toxicological Unit of Careggi General Hospital (University of Florence, Italy) for amatoxin poisoning. Data included the biological parameters monitored, the treatment protocols used (intensive fluid and supportive therapy, restitution of the altered coagulation factors, multiple-dose activated charcoal, mannitol, dexamethasone, glutathione, and penicillin G), and outpatient follow-up evaluations. RESULTS: The clinical data of 111 patients were evaluated; their biological parameters were monitored every 12-24 hours until discharge. Two patients died; both were admitted to the hospital more than 60 hours after mushroom ingestion. Of all the laboratory parameters evaluated, the evolution of hepatic transaminases and prothrombin activity over four days were the most predictive indicators of recovery or death. Our follow-up evaluation of 105 patients demonstrated that our survivors recovered completely. CONCLUSIONS: Our experience indicates that the protocol used in our Toxicologicy Unit is effective for amatoxin poisoning, and that all patients treated within 36 hours after mushroom ingestion were cured without sequelae.
Assuntos
Amanitinas/intoxicação , Intoxicação Alimentar por Cogumelos/terapia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Alanina Transaminase/sangue , Amanitinas/urina , Aspartato Aminotransferases/sangue , Feminino , Seguimentos , Hospitalização , Humanos , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Intoxicação Alimentar por Cogumelos/sangue , Intoxicação Alimentar por Cogumelos/epidemiologia , Intoxicação Alimentar por Cogumelos/urina , Prognóstico , Tempo de Protrombina , Fatores de TempoRESUMO
Amine oxidases (AOs) are ubiquitous enzymes involved in the metabolism of biogenic amines. Copper AOs (Cu-AOs) catalyze the oxidative deamination of primary amine groups of several biogenic amines, such as putrescine, cadaverine, and histamine. In the present review, the effects of a plant amine oxidase (Cu-AO, histaminase, EC1.4.3.6) purified from pea seedlings in the modulation of IgE-mediated allergic reactions, and in the prevention of cardiac and splachnic postischemic reperfusion damage are reported.
Assuntos
Amina Oxidase (contendo Cobre)/uso terapêutico , Anafilaxia/tratamento farmacológico , Asma/tratamento farmacológico , Isquemia Encefálica/tratamento farmacológico , Hipersensibilidade/tratamento farmacológico , Inflamação/tratamento farmacológico , Pisum sativum/enzimologia , Isquemia Encefálica/complicações , Humanos , Fitoterapia/métodos , Fitoterapia/tendências , Extratos Vegetais/uso terapêutico , Guias de Prática Clínica como Assunto , Padrões de Prática Médica , Sementes/enzimologia , Choque/etiologia , Choque/prevenção & controleRESUMO
Reactive oxygen species have been implicated in the pathogenesis of asthma and, in atopic asthmatics, endogenous superoxide dismutase (SOD) enzyme levels are known to decrease. This suggests that replacing a failed endogenous SOD enzyme system with a mimetic of the endogenous enzyme would be beneficial and protective. In this study we demonstrate that removal of superoxide by the SOD mimetic (SODm) M40403 reduces the respiratory and histopathological lung abnormalities due to ovalbumin (OA) aerosol in a model of allergic asthma-like reaction in sensitized guinea pigs. Both respiratory abnormalities and bronchoconstriction in response to OA challenge are nearly absent in naïve animals, while they sharply became severe in sensitized animals. In addition, OA aerosol induced a reduction of MnSOD activity which was paralleled with bronchiolar lumen reduction, pulmonary air space hyperinflation, mast cell degranulation, eosinophil infiltration, bronchial epithelial cell apoptosis, increase in myeloperoxidase activity, malonyldialdehyde production and 8-hydroxy-2'-deoxyguanosine formation in the lung tissue, as well as elevation of PGD2 in the bronchoalveolar lavage fluid. Treatment with M40403 restored the levels of MnSOD activity and significantly reduced all the above parameters. In summary, our findings support the potential therapeutic use of SOD mimetics in asthma and anaphylactic reactions and account for a critical role for superoxide in acute allergic asthma-like reaction in actively sensitized guinea pig.
Assuntos
Asma/tratamento farmacológico , Compostos Organometálicos/uso terapêutico , 8-Hidroxi-2'-Desoxiguanosina , Alérgenos , Animais , Apoptose/efeitos dos fármacos , Asma/induzido quimicamente , Líquido da Lavagem Broncoalveolar/química , Broncoconstrição/efeitos dos fármacos , Desoxiguanosina/análogos & derivados , Desoxiguanosina/antagonistas & inibidores , Cobaias , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Malondialdeído/metabolismo , Manganês , Ovalbumina , Peroxidase/metabolismo , Prostaglandina D2/análise , Superóxido Dismutase/metabolismo , Superóxido Dismutase/uso terapêuticoRESUMO
1. Myocardial injury caused by ischaemia and reperfusion comes from multiple pathogenic events, including endothelial damage, neutrophil extravasation into tissue, mast cell activation, and peroxidation of cell membrane lipids. These events are followed by myocardial cell alterations resulting eventually in cell necrosis. An enhanced formation of reactive oxygen species is widely accepted as a stimulus for tissue destruction and cardiac failure. 2. In this study, we have investigated the cardioprotective effects of M40403 in myocardial ischaemia-reperfusion injury. M40403 is a low molecular weight, synthetic manganese containing superoxide dismutase mimetic (SODm) that selectively removes superoxide anion. Ischaemia was induced in rat hearts in vivo by ligating the left anterior descending coronary artery. Thirty minutes after the induction of ischaemia, the ligature was removed and reperfusion allowed to occur for at least 60 min. M40403 (0.1-1 mg kg(-1)) was given intravenously 15 min before ischaemia. 3. The results obtained in this study showed that M40403 significantly reduced the extent of myocardial damage, mast cell degranulation and the incidence of ventricular arrhythmias. Furthermore, M40403 significantly attenuated, in a dose-dependent manner, neutrophil infiltration in the myocardium as well as the associated induction of lipid peroxidation. Calcium overload seen post-reperfusion of the ischaemic myocardium was also reduced by M40403. 4. Immunohistochemical analysis for nitrotyrosine revealed a positive staining in cardiac tissue taken after reperfusion: this was attenuated by M40403. Moreover reperfused cardiac tissue sections showed positive staining for P-selectin and for anti-intercellular adhesion molecule (ICAM-1) in the vascular endothelial cells. M40403 treatment markedly reduced the intensity and degree of P-selectin and ICAM-1 in these tissues. No staining for nitrotyrosine, P-selectin or ICAM-1 was found in cardiac tissue taken at the end of the ischaemic period. 5. Overall, M40403 treatment reduced the morphological signs of myocardial cell injury and significantly improved survival. 6. Taken together, these results clearly indicate that M40403 treatment exerts a protective effect against ischaemia-reperfusion-induced myocardial injury, supporting a key role for superoxide anion in reperfusion injuries. This suggests that synthetic enzymes of SOD such as M40403, offer a novel therapeutic approach for the treatment of ischaemic heart disease where superoxide anion plays a dominant role.
Assuntos
Cardiotônicos/farmacologia , Manganês , Isquemia Miocárdica/prevenção & controle , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Compostos Organometálicos/farmacologia , Superóxido Dismutase/química , Tirosina/análogos & derivados , Animais , Cardiotônicos/uso terapêutico , Imuno-Histoquímica , Molécula 1 de Adesão Intercelular/metabolismo , Masculino , Mimetismo Molecular , Isquemia Miocárdica/metabolismo , Isquemia Miocárdica/patologia , Traumatismo por Reperfusão Miocárdica/metabolismo , Traumatismo por Reperfusão Miocárdica/patologia , Compostos Organometálicos/química , Compostos Organometálicos/uso terapêutico , Selectina-P/metabolismo , Peroxidase/metabolismo , Ratos , Ratos Wistar , Tirosina/metabolismoRESUMO
We report the effects of exogenous and endogenous carbon monoxide (CO) on the immunological activation of human basophils. Hemin (1-100 microM), a heme oxygenase substrate analogue, significantly increased the formation of bilirubin from partially purified human basophils, thus indicating that these cells express heme oxygenase. This effect was reversed by preincubating the cells for 30 min with Zn-protoporphyrin IX (100 microM), a heme oxygenase inhibitor. Hemin (100 microM) also decreased immunoglobulin G anti-Fcepsilon (anti-IgE)-induced activation of basophils, measured by the expression of a membrane granule-associated protein, identified as cluster differentiation protein 63 (CD63), and by histamine release. These effects were reversed by Zn-protoporphyrin IX (100 microM), by oxyhemoglobin (HbO(2)), a CO scavenger (100 microM), and by 1H-[1,2,4]oxadiazolo[4,3-a] quinoxalin-1-one (ODQ), an inhibitor of the soluble guanylyl cyclase (100 microM). Exposure of basophils to exogenous CO (10 microM for 30 min) also decreased their activation, while nitrogen (N(2)) was ineffective. HbO(2) and ODQ reversed the inhibition, reversing both membrane protein CD63 expression and histamine release to basal values. Both hemin and exogenous CO significantly raised cGMP levels in basophils and blunted the rise of calcium levels caused by immunological activation. This study suggests that CO increases cGMP formation, which in turn induces a fall in intracellular Ca(2+) concentration, thereby resulting in the inhibition of human basophil activation.
Assuntos
Basófilos/metabolismo , Monóxido de Carbono/farmacologia , Heme Oxigenase (Desciclizante)/metabolismo , Receptores de IgE/metabolismo , Anticorpos Anti-Idiotípicos/farmacologia , Antígenos CD/biossíntese , Basófilos/imunologia , Cálcio/metabolismo , Monóxido de Carbono/metabolismo , GMP Cíclico/metabolismo , Citometria de Fluxo , Guanilato Ciclase , Heme Oxigenase (Desciclizante)/antagonistas & inibidores , Heme Oxigenase (Desciclizante)/biossíntese , Hemina/farmacologia , Liberação de Histamina , Humanos , Técnicas In Vitro , Microscopia Eletrônica , Oxidiazóis/farmacologia , Glicoproteínas da Membrana de Plaquetas/biossíntese , Protoporfirinas/farmacologia , Receptores Citoplasmáticos e Nucleares/antagonistas & inibidores , Receptores de IgE/biossíntese , Receptores de IgE/imunologia , Guanilil Ciclase Solúvel , Tetraspanina 30RESUMO
This study evaluates the effects of a copper amine oxidase (histaminase) purified from the pea seedling as a free or immobilized enzyme on asthmalike reactions to inhaled antigen in actively sensitized guinea pig in vivo. Male albino guinea pigs, sensitized with ovalbumin, were challenged with the antigen given by aerosol; free histaminase or CNBr-Sepharose immobilized histaminase was given intraperitoneally (20 microg, 3 or 24 h before antigen challenge) or by aerosol (4 microg, 30 min before or during ovalbumin aerosol). The following parameters were examined: latency time for the onset of respiratory abnormalities, cough severity score, and occurrence and duration of dyspnea. We also evaluated lung histopathology, mast cell degranulation, and lung myeloperoxidase and malonydialdehyde levels. Histaminase significantly reduced the severity of cough and the occurrence of dyspnea and delayed the onset of respiratory abnormalities. Both enzymes prevented bronchial constriction, pulmonary air space inflation, leukocyte infiltration (evaluated as myeloperoxidase activity), and lipoperoxidation of cell membranes (evaluated as malonyldialdehyde production). No relevant differences in pharmacological potency were noted between free or immobilized enzyme. This study provides evidence that histaminase counteracts acute allergic asthmalike reaction in actively sensitized guinea pigs, raising the possibility of new therapeutic strategies for allergic asthma in humans.
Assuntos
Amina Oxidase (contendo Cobre)/uso terapêutico , Antígenos/administração & dosagem , Pisum sativum , Hipersensibilidade Respiratória/tratamento farmacológico , Hipersensibilidade Respiratória/enzimologia , Administração por Inalação , Aerossóis , Amina Oxidase (contendo Cobre)/isolamento & purificação , Amina Oxidase (contendo Cobre)/farmacologia , Animais , Asma/tratamento farmacológico , Asma/enzimologia , Asma/imunologia , Tosse/tratamento farmacológico , Tosse/enzimologia , Tosse/imunologia , Enzimas Imobilizadas/sangue , Cobaias , Imunização/métodos , Injeções Intraperitoneais , Pulmão/efeitos dos fármacos , Pulmão/enzimologia , Masculino , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Testes de Função Respiratória , Hipersensibilidade Respiratória/imunologia , PlântulaRESUMO
This study investigates whether relaxin (RLX), a hormone previously shown to inhibit mast cell function and to stimulate endogenous nitric oxide (NO) biosynthesis, counteracts the activation of isolated human basophils stimulated with anti-IgE or phorbol ester, and, if so, whether NO is involved. RLX reduced dose-dependently the expression of the activation marker CD63, the release of histamine and the rise of intracellular Ca2+ levels which triggers granule release by stimulated basophils. RLX also blunts the ultrastructural signs of anaphylactic granule release. The effects of RLX appear to depend upon activation of Ca2+/calmodulin-dependent NO synthase and endogenous NO production. They were reproduced by the NO donor sodium nitroprusside (SNP) and were reverted by the NO synthase inhibitor N(omega)-monomethyl-L-arginine, or by the NO scavenger oxyhemoglobin, or by blocking the NO physiological target guanylate cyclase with ODQ. In conclusion, RLX appears to play a role in down-regulating basophil function upon immunologic and nonimmunologic activation.
Assuntos
Basófilos/efeitos dos fármacos , Basófilos/metabolismo , Óxido Nítrico/fisiologia , Receptores de IgE/biossíntese , Relaxina/farmacologia , Basófilos/imunologia , Humanos , Óxido Nítrico/imunologia , Receptores de IgE/imunologiaRESUMO
Cardiac anaphylaxis refers to the functional and metabolic changes in the heart caused by the anaphylactic release of histamine and vasoactive products of arachidonic acid cascade by mast cells and basophils. As in most type I hypersensitivity-based diseases, histamine plays a key role in the pathophysiology of cardiac anaphylaxis. In the heart, mast cell activation and histamine release are controlled by multiple endogenous mechanisms, including adrenergic neural control, histamine-dependent negative feedback operated through H2 receptors, and the endogenous generation of nitric oxide (NO) and carbon monoxide (CO). All these mechanisms can be targeted by substances that have revealed a clear-cut effect in blunting cardiac anaphylaxis in experimental animal models, and could be developed as potential, novel anti-anaphylactic drugs. In this article, we discuss new findings and significant trends related to this topic.
Assuntos
Anafilaxia/imunologia , Cardiopatias/imunologia , Histamina/imunologia , Amina Oxidase (contendo Cobre)/uso terapêutico , Anafilaxia/terapia , Animais , Monóxido de Carbono/fisiologia , Cardiopatias/terapia , Antagonistas dos Receptores Histamínicos/uso terapêutico , Liberação de Histamina/fisiologia , Humanos , Mastócitos/efeitos dos fármacos , Mastócitos/fisiologia , Óxido Nítrico/fisiologia , Relaxina/uso terapêuticoRESUMO
Cannabinoids exert complex actions on neurotransmitter systems involved in cognition, locomotion, appetite, but no information was available so far on the interactions between the endocannabinoid system and histaminergic neurons that command several, similar behavioural states and memory. In this study, we investigated the effect of cannabimimetic compounds on histamine release using the microdialysis technique in the brain of freely moving rats. We found that systemic administration of the cannabinoid receptors 1 (CB1-r) agonist arachidonyl-2'chloroethylamide/N-(2chloroethyl)-5Z,8Z,11Z,14Z-eicosatetraenamide (ACEA; 3 mg/kg) increased histamine release from the posterior hypothalamus, where the histaminergic tuberomamillary nuclei (TMN) are located. Local infusions of ACEA (150 nm) or R(+)-methanandamide (mAEA; 1 microm), another CB1-r agonist, in the TMN augmented histamine release from the TMN, as well as from two histaminergic projection areas, the nucleus basalis magnocellularis and the dorsal striatum. When the endocannabinoid uptake inhibitor AM404 was infused into the TMN, however, increased histamine release was observed only in the TMN. The cannabinoid-induced effects on histamine release were blocked by co-administrations with the CB1-r antagonist AM251. Using double-immunofluorescence labelling and confocal laser-scanning microscopy, CB1-r immunostaining was found in the hypothalamus, but was not localized onto histaminergic cells. The modulatory effect of cannabimimetic compounds on histamine release apparently did not involve inhibition of gamma-aminobutyric acid (GABA)ergic neurotransmission, which provides the main inhibitory input to the histaminergic neurons in the hypothalamus, as local infusions of ACEA did not modify GABA release from the TMN. These profound effects of cannabinoids on histaminergic neurotransmission may partially underlie some of the behavioural changes observed following exposure to cannabinoid-based drugs.
Assuntos
Ácidos Araquidônicos/farmacologia , Moduladores de Receptores de Canabinoides/farmacologia , Canabinoides/agonistas , Diferenciação Celular/efeitos dos fármacos , Endocanabinoides , Histamina/metabolismo , Hipotálamo/efeitos dos fármacos , Análise de Variância , Animais , Bicuculina/farmacologia , Diferenciação Celular/fisiologia , Cromatografia Líquida de Alta Pressão/métodos , Antagonistas GABAérgicos/farmacologia , Imuno-Histoquímica/métodos , Masculino , Microdiálise/métodos , Piperidinas/farmacologia , Pirazóis/farmacologia , Ratos , Ratos Sprague-Dawley , Receptor CB1 de Canabinoide/metabolismo , Fatores de Tempo , Vigília , Ácido gama-Aminobutírico/metabolismoRESUMO
The basolateral amygdala (BLA) is involved in learning that certain environmental cues predict threatening events. Several studies have shown that manipulation of neurotransmission within the BLA affects the expression of memory after fear conditioning. We previously demonstrated that blockade of histaminergic H3 receptors decreased spontaneous release of acetylcholine (ACh) from the BLA of freely moving rats, and impaired retention of fear memory. In the present study, we examined the effect of activating H3 receptors within the BLA on both ACh release and expression of fear memory. Using the microdialysis technique in freely moving rats, we found that the histaminergic H3 agonists R-alpha-methylhistamine (RAMH) and immepip, directly administered into the BLA, augmented spontaneous release of ACh in a similar manner. Levels of ACh returned to baseline on perfusion with control medium. Rats receiving intra-BLA, bilateral injections of the H3 agonists at doses similar to those enhancing ACh spontaneous release, immediately after contextual fear conditioning, showed stronger memory for the context-footshock association, as demonstrated by longer freezing assessed at retention testing performed 72 h later. Post-training, bilateral injections of 15 ng oxotremorine also had a similar effect on memory retention, supporting the involvement of the cholinergic system. Thus, our results further support a physiological role for synaptically released histamine, that in addition to affecting cholinergic transmission in the amygdala, modulates consolidation of fear memories
Assuntos
Acetilcolina/metabolismo , Tonsila do Cerebelo/metabolismo , Medo/fisiologia , Histamina/metabolismo , Memória/fisiologia , Neurônios/metabolismo , Oxotremorina/análogos & derivados , Receptores Histamínicos H3/metabolismo , Tonsila do Cerebelo/efeitos dos fármacos , Animais , Inibidores da Colinesterase/farmacologia , Condicionamento Psicológico/efeitos dos fármacos , Condicionamento Psicológico/fisiologia , Relação Dose-Resposta a Droga , Medo/efeitos dos fármacos , Agonistas dos Receptores Histamínicos/farmacologia , Imidazóis/farmacologia , Masculino , Memória/efeitos dos fármacos , Metilistaminas/farmacologia , Agonistas Muscarínicos/farmacologia , Neurônios/efeitos dos fármacos , Oxotremorina/farmacologia , Piperidinas/farmacologia , Ratos , Ratos Wistar , Receptores Histamínicos H3/efeitos dos fármacos , Transmissão Sináptica/efeitos dos fármacos , Transmissão Sináptica/fisiologiaRESUMO
The effects of histaminergic ligands on both ACh spontaneous release from the hippocampus and the expression of c-fos in the medial septum-diagonal band (MSA-DB) of freely moving rats were investigated. Because the majority of cholinergic innervation to the hippocampus is provided by MSA-DB neurons, we used the dual-probe microdialysis technique to apply drugs to the MSA-DB and record the induced effects in the projection area. Perfusion of MSA-DB with high-KCl medium strongly stimulated hippocampal ACh release which, conversely, was significantly reduced by intra-MSA-DB administration of tetrodotoxin. Histamine or the H2 receptor agonist dimaprit, applied directly to the hippocampus, failed to alter ACh release. Conversely, perfusion of MSA-DB with these two compounds increased ACh release from the hippocampus. Also, thioperamide and ciproxifan, two H3 receptor antagonists, administered into MSA-DB, increased the release of hippocampal ACh, whereas R-alpha-methylhistamine, an H3 receptor agonist, produced the opposite effect. The blockade of MSA-DB H2 receptors, caused by local perfusion with the H2 receptor antagonist cimetidine, moderated the spontaneous release of hippocampal ACh and antagonized the facilitation produced by H3 receptor antagonists. Triprolidine, an H1 receptor antagonist, was without effect. Moreover, cells expressing c-fos immunoreactivity were significantly more numerous in ciproxifan- or thioperamide-treated rats than in controls, although no colocalization of anti-c-fos and anti-ChAT immunoreactivity was observed. These results indicate a role for endogenous histamine in modulating the cholinergic tone in the hippocampus.
Assuntos
Acetilcolina/metabolismo , Hipocampo/metabolismo , Histamina/fisiologia , Núcleos Septais/metabolismo , Animais , Agonistas dos Receptores Histamínicos/administração & dosagem , Agonistas dos Receptores Histamínicos/farmacologia , Antagonistas dos Receptores Histamínicos/administração & dosagem , Antagonistas dos Receptores Histamínicos/farmacologia , Antagonistas dos Receptores Histamínicos H1/farmacologia , Imidazóis/farmacologia , Masculino , Microdiálise , Neurônios/metabolismo , Perfusão , Piperidinas/farmacologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Wistar , Receptores Histamínicos H3/efeitos dos fármacos , Núcleos Septais/citologia , Tetrodotoxina/administração & dosagem , Tetrodotoxina/farmacologia , Triprolidina/farmacologiaRESUMO
The effect of a copper amine oxidase (histaminase) purified from the pea seedling as free or immobilized enzyme on the response to specific antigen was studied in isolated hearts from actively sensitized guinea pigs. In vitro challenge with the specific antigen of hearts from actively sensitized animals evokes a positive inotropic and chronotropic effect, a coronary constriction, followed by dilation and an increase in the amount of histamine and nitrites, the oxidation product of nitric oxide, in the perfusates. In the presence of both forms of histaminases, the positive inotropic and chronotropic responses as well as the coronary constriction and the release of histamine were fully blocked. The amount of nitrites, appearing in the perfusates when anaphylaxis is elicited in the presence of both forms of histaminases, is significantly increased, as well as nitric oxide synthase activity and cyclic GMP content in cardiac tissue, while cardiac calcium overload was significantly prevented. These observations demonstrate that the decrease in the anaphylactic release of histamine and the subsequent abatement of the cardiac response to antigen can be accounted for by the inactivation by histaminase of the released histamine and by a stimulation of endogenous nitric oxide production.
Assuntos
Amina Oxidase (contendo Cobre)/metabolismo , Amina Oxidase (contendo Cobre)/farmacologia , Anafilaxia/enzimologia , Animais , Cálcio/metabolismo , Computadores , GMP Cíclico/metabolismo , Densitometria , Feminino , Cobaias , Histamina/metabolismo , Masculino , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase/metabolismo , Pisum sativum/enzimologia , Extratos Vegetais/farmacologia , Fatores de TempoRESUMO
Grass pea seedling histaminase (a copper-diamine oxidase) was found to exert a significant cardioprotection against post-ischaemic reperfusion damage. Electrocardiogram (ECG) recordings from the rats subjected in vivo to ischaemia and reperfusion showed ventricular tachycardia (VT) and ventricular fibrillations (VF) occurring in 9 out of 12 untreated rats whereas no ventricular arrhythmias were found under histaminase (80U/kg body weight) treatment (n=16 rats). Computer-assisted morphometry of the ischaemic reperfused hearts stained with nitroblue tetrazolium showed the extension of damaged myocardium (area at risk and infarct size) significantly reduced in rats treated with histaminase, in comparison with the non-treated rats, whereas no protection was found with the semicarbazide inactivated histaminase. Biochemical markers of ischaemia-reperfusion myocardial tissue damage: malonyldialdehyde (MDA), tissue calcium concentration, myeloperoxidase (MPO), and apoptosis indicator caspase-3 were significantly elevated in untreated post-ischaemic reperfused rats, but significantly reduced under histaminase protection. In conclusion, plant histaminase appears to protect hearts from ischaemia-reperfusion injury by more than one mechanism, essentially involving histamine oxidation, and possibly as reactive oxygen species scavenger, presenting good perspectives for a novel therapeutic approach in treatment of ischaemic heart pathology.
Assuntos
Amina Oxidase (contendo Cobre)/uso terapêutico , Isquemia Miocárdica/diagnóstico , Isquemia Miocárdica/tratamento farmacológico , Traumatismo por Reperfusão Miocárdica/diagnóstico , Traumatismo por Reperfusão Miocárdica/tratamento farmacológico , Amina Oxidase (contendo Cobre)/isolamento & purificação , Amina Oxidase (contendo Cobre)/metabolismo , Animais , Cardiotônicos/farmacologia , Masculino , Isquemia Miocárdica/complicações , Traumatismo por Reperfusão Miocárdica/etiologia , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Pisum sativum/metabolismo , Ratos , Ratos Wistar , Sementes/metabolismo , Resultado do TratamentoRESUMO
The antigen-induced release of histamine from sensitized guinea pig mast cells was dose-dependently reduced by endogenous (2-arachidonylglycerol; 2AG) and exogenous [(1R,3R,4R)-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-4-(3-hydroxypropyl)cyclohexan-1-ol (CP55,940)] cannabinoids. The inhibitory action afforded by 2AG and CP55,940 was reversed by N-[(1S)-endo-1,3,3-trimethylbicyclo[2.2.1]heptan-2-yl]5-(4-chloro-3-methylphenyl)-1-(4-methylbenzyl)pyrazole-3-carboxamide (SR144528), a selective cannabinoid 2 (CB(2)) receptor antagonist, and left unchanged by the selective CB(1) antagonist N-(piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (AM251). The inhibitory action of 2AG and CP55,940 was reduced by the unselective nitric-oxide synthase (NOS) inhibitor N-monomethyl-L-arginine methylester (l-NAME) and reinstated by L-arginine, the physiological substrate. The inhibitory action of 2AG and CP55,940 was also reduced by the unselective cyclooxygenase (COX) inhibitor indomethacin and the selective COX-2 blocker rofecoxib. Both 2AG and CP55,940 significantly increased the production of nitrite from mast cells, which was abrogated by L-NAME and N-(3-(aminomethyl)benzyl)acetamidine (1400W), a selective inducible NOS (iNOS) inhibitor. Nitrite production consistently paralleled a CP55,940-induced increase in the expression of iNOS protein in mast cells. Both 2AG and CP55,940 increased the generation of prostaglandin E(2) from mast cells, which was abrogated by indomethacin and rofecoxib and parallel to the CP55,940-induced expression of COX-2 protein. Mast cell challenge with antigen was accompanied by a net increase in intracellular calcium levels. Both cannabinoid receptor ligands decreased the intracellular calcium levels, which were reversed by SR144528 and l-NAME. In unstimulated mast cells, both ligands increased cGMP levels. The increase was abrogated by SR144528, l-NAME, indomethacin, and rofecoxib. Our results suggest that 2AG and CP55,940 decreased mast cell activation in a manner that is susceptible to a CB(2) receptor antagonist and to inhibition of nitric oxide and prostanoid pathways.
Assuntos
Ácidos Araquidônicos/farmacologia , Moduladores de Receptores de Canabinoides/farmacologia , Eicosanoides/fisiologia , Endocanabinoides , Glicerídeos/farmacologia , Imunidade/efeitos dos fármacos , Mastócitos/efeitos dos fármacos , Óxido Nítrico/fisiologia , Animais , Western Blotting , Cicloexanóis/farmacologia , Inibidores Enzimáticos/farmacologia , Cobaias , Imunossupressores/farmacologia , Masculino , Mastócitos/imunologia , NG-Nitroarginina Metil Éster/farmacologia , Receptor CB2 de Canabinoide/metabolismoRESUMO
In a previous study, it was found that the decrease in the total plasma bilirubin level (Btot) in preterm infants was associated with the decrease in oxidative stress. We hypothesized that this occurs as a result of a pro-oxidant effect of heme oxygenase (HO), which outcompetes with the antioxidant properties of bilirubin. In this study we studied 12 preterm infants in whom the plasma levels of Btot, total hydroperoxide (TH), protein SH groups, HO activity, non-transferrin-bound iron (NTBI), and erythrocyte CuZn superoxide dismutase (CuZn SOD) activity were concurrently measured when the Btot was >220 microM and after a Btot drop of >34 microM. The Btot decrease was concurrent with the TH decrease, protein SH groups increase, and the HO and CuZn SOD activity increase and was not associated with an NTBI increase. We concluded that 1) Btot does not exert a meaningful antioxidant effect in vivo; 2) HO does not exert a pro-oxidant effect involving an NTBI increase and that, on the contrary, it could exert an antioxidant effect; and 3) the concurrent HO and CuZn SOD activity increase could indicate a synergic antioxidant effect of the two enzymes.