RESUMO
Oxidation of lipoprotein-derived lipids is generally accepted to be important in atherogenesis, and lipophilic antioxidants have been suggested as potential antiatherosclerotic agents. The antiatherogenic effects observed by certain antioxidants, especially probucol, in different animal models support this suggestion. There are however also cases where other lipophilic antioxidants have not been able to support this hypothesis. This has raised the question whether the effects of probucol and similar compounds are mainly due to some other property, unrelated to their antioxidant efficacy. For example, probucol is shown to possess immunomodulatory properties. Immune reactions are known to occur during atherogenesis. We therefore tested the dimer of N-acetylcysteine, DiNAC, which is a disulfide with immunomodulating properties and enhances oxazolone-induced contact sensitivity (CS) reactions in mice, for effects on atherosclerosis. When given to male heritable hyperlipidemic rabbit (WHHL) rabbits from 10 to 22 weeks of age, this compound reduced by 50% thoracic aorta atherosclerosis (p < 0.05), without affecting plasma lipid levels. Here we also show that probucol and a close chemical analog, both known to prevent atherosclerosis in WHHL rabbits, enhance the CS reaction in mice, while two other related antioxidants did not affect the CS reaction. At least one of these is also without effect on atherosclerosis in WHHL rabbits. The results show that DiNAC might represent a new treatment modality for atherosclerosis-related disease, and suggest that some antioxidants may have antiatherosclerotic properties more related to "immunomodulatory" properties than to antioxidant properties in general.
Assuntos
Acetilcisteína/química , Adjuvantes Imunológicos/farmacologia , Arteriosclerose/prevenção & controle , Cistina/análogos & derivados , Cistina/farmacologia , Adjuvantes Imunológicos/química , Animais , Antioxidantes/química , Arteriosclerose/genética , Arteriosclerose/patologia , Colesterol/sangue , Cistina/química , Dermatite de Contato/imunologia , Dissulfetos/química , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Oxazolona/farmacologia , Probucol/farmacologia , Coelhos , Receptores de LDL/deficiência , Receptores de LDL/genética , Relação Estrutura-Atividade , Triglicerídeos/sangueRESUMO
Oral N-acetyl-L-cysteine (NAC) is used clinically for treatment of chronic obstructive pulmonary disease. NAC is easily oxidized to its disulfide. We show here that N,N'-diacetyl-L-cystine (DiNAC) is a potent modulator of contact sensitivity (CS)/delayed type hypersensitivity (DTH) reactions in rodents. Oral treatment of BALB/c mice with 0.003 to 30 micromol/kg DiNAC leads to enhancement of a CS reaction to oxazolone; DiNAC is 100 to 1000 times more potent than NAC in this respect, indicating that it does not act as a prodrug of NAC. Structure-activity studies suggest that a stereochemically-defined disulfide element is needed for activity. The DiNAC-induced enhancement of the CS reaction is counteracted by simultaneous NAC-treatment; in contrast, the CS reaction is even more enhanced in animals treated with DiNAC together with the glutathione-depleting agent buthionine sulfoximine. These data suggest that DiNAC acts via redox processes. Immunohistochemically, ear specimens from oxazolone-sensitized and -challenged BALB/c mice treated with DiNAC display increased numbers of CD8(+) cells. DiNAC treatment augments the CS reaction also when fluorescein isothiocyanate is used as a sensitizer in BALB/c mice; this is a purported TH2 type of response. However, when dinitrofluorobenzene is used as a sensitizer, inducing a purported TH1 type of response, DiNAC treatment reduces the reaction. Treatment with DiNAC also reduces a DTH footpad-swelling reaction to methylated BSA. Collectively, these data indicate that DiNAC in vivo acts as a potent and effective immunomodulator that can either enhance or reduce the CS or DTH response depending on the experimental conditions.
Assuntos
Adjuvantes Imunológicos/farmacologia , Cistina/análogos & derivados , Dermatite de Contato/imunologia , Hipersensibilidade Tardia/imunologia , Acetilcisteína/análogos & derivados , Animais , Linfócitos T CD8-Positivos , Cistina/farmacologia , Dermatite de Contato/etiologia , Dinitrofluorbenzeno , Orelha , Feminino , Fluoresceína-5-Isotiocianato , Pé , Granuloma/etiologia , Granuloma/imunologia , Hipersensibilidade Tardia/etiologia , Imuno-Histoquímica , Contagem de Linfócitos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos , Oxazolona , Coelhos , Soroalbumina BovinaRESUMO
Five stereochemically constrained analogs of the chemotactic tripeptide incorporating 1-aminocycloalkane-1-carboxylic acid (Ac(n)c) and alpha,alpha-dialkylglycines (Deg, diethylglycine; Dpg, n,n-dipropylglycine and Dbg, n,n-dibutylglycine) at position 2 have been synthesized. NMR studies of peptides For-Met-Xxx-Phe-OMe (Xxx=Ac(7)c, I; Ac(8)c, II; Deg, III; Dpg, IV and Dbg, V; For, formyl) establish that peptides with cycloalkyl residues, I and II, adopt folded beta-turn conformations in CDCl3 and (CD3)2SO. In contrast, analogs with linear alkyl sidechains, III-V, favour fully extended (C5) conformations in solution. Peptides I-V exhibit high activity in inducing beta-glucosaminidase release from rabbit neutrophils, with ED50 values ranging from 1.4-8.0 x 10(-11) M. In human neutrophils the Dxg peptides III-V have ED50 values ranging from 2.3 x 10(-8) to 5.9 x 10(-10) M, with the activity order being V > IV > III. While peptides I-IV are less active than the parent, For-Met-Leu-Phe-OH, in stimulating histamine release from human basophils, the Dbg peptide V is appreciably more potent, suggesting its potential utility as a probe for formyl peptide receptors.
Assuntos
Aminoácidos/química , Hexosaminidases/metabolismo , Liberação de Histamina/efeitos dos fármacos , Neutrófilos/metabolismo , Oligopeptídeos/química , Sequência de Aminoácidos , Animais , Relação Dose-Resposta a Droga , Hexosaminidases/efeitos dos fármacos , Humanos , Espectroscopia de Ressonância Magnética , Neutrófilos/efeitos dos fármacos , Oligopeptídeos/farmacologia , Coelhos , Estereoisomerismo , Relação Estrutura-AtividadeRESUMO
To assess the role of protein kinase C (PKC) in the respiratory burst of adherent human polymorphonuclear leukocytes (PMNL), reduction of ferricytochrome C by cells triggered with a phorbol ester (PMA), ionophore A23187, serum-treated zymosan (STZ) or three lipid derivatives, 3-decanoyl-sn-glycerol (G-3-OCOC9), (R,R)-1,4-diethyl-2-O-decyl-L-tartrate (Tt-2-OC10) and 3-decyloxy-5-hydroxymethylphenol (DHP) was examined in a microtiter plate procedure in the presence of inhibitors of PKC and, for comparison, inhibitors of calmodulin, diacylglycerol and myosin light chain kinases and the peptidyl-prolyl cis-trans isomerase activity of fujiphilin. 1) Of the protein kinase inhibitors examined, Ro 31-7549 and staurosporine reduced responses to all stimuli except possibly STZ; in contrast, K252a and the myosin light chain kinase inhibitors ML-7 and ML-9 blocked responses to A23187 and STZ better than those triggered by PMA. H-7 reduced responses to A23187, DHP and G-3-OCOC9, and calphostin, palmitoyl carnitine, sphingosine and the multifunctional drugs TMB-8 and W-7 reduced A23187; they also, when examined, reduced decane derivative-induced O2- production more effectively than PMA- and STZ-triggered responses. Polymyxin B, 4 alpha-PMA and retinal displayed no inhibitory capacity. 2) Of the selective calmodulin antagonists, CGS 9343B, Ro 22-4839 and calmidazolium did not inhibit the oxidative response irrespective of the stimulus used, whereas metofenazate reduced those evoked by A23187, DHP, G-3-OCOC9 and STZ.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Isomerases de Aminoácido/antagonistas & inibidores , Calmodulina/antagonistas & inibidores , Proteínas de Transporte/antagonistas & inibidores , Quinase de Cadeia Leve de Miosina/antagonistas & inibidores , Neutrófilos/metabolismo , Fosfotransferases/antagonistas & inibidores , Proteína Quinase C/antagonistas & inibidores , Superóxidos/metabolismo , Basófilos/efeitos dos fármacos , Basófilos/metabolismo , Carbazóis/farmacologia , Diacilglicerol Quinase , Liberação de Histamina/efeitos dos fármacos , Humanos , Alcaloides Indólicos , Neutrófilos/efeitos dos fármacos , Peptidilprolil Isomerase , Pirimidinonas/farmacologia , Tacrolimo/farmacologia , Acetato de Tetradecanoilforbol/farmacologia , Tiazóis/farmacologiaRESUMO
To assess the role of calmodulin in human basophil histamine release, we triggered leukocytes with different secretagogues in the presence of putative inhibitors of calmodulin. Calcium ionophore-induced histamine release was reduced or blocked by calmidazolium, CGS 9343B, felodipine, metofenazate, and Ro 22-4839. H 186/86, a felodipine-related dihydropyridine derivative, blocked A23187-but not ionomycin-triggered histamine release, suggesting a difference in the mode of action of these ionophores. In contrast, leukocyte histamine release triggered by the purported protein kinase C (PKC) activator, 1,2-isopropylidene-3-decanoyl-sn-glycerol (IpOCOC9), was enhanced by calmidazolium, CGS 9349B and metofenazate but not affected by felodipine or Ro 22-4839, whereas the response triggered by 4 beta-phorbol 12-myristate 13-acetate (PMA) was reduced by metofenazate and Ro 22-4839 but not consistently affected by calmidazolium, CGS 9343B or felodipine. The PMA-induced histamine release was enhanced by H 186/86. Anti-IgE- and FMLP-induced responses were either unaffected or slightly enhanced by the examined calmodulin antagonists. In comparison with the calmodulin antagonists, R 59022, an inhibitor of diacylglycerol kinase, failed to reduce calcium ionophore-triggered histamine release, whereas FK506, an inhibitor of peptidyl prolyl cis-trans isomerase (PPI), reduced both anti-IgE- and ionophore-triggered responses. These results indicate that calmodulin constitutes an obligate link in signal transduction pathways leading to human leukocyte histamine release if the trigger is a calcium ionophore but not when responses are induced by anti-IgE, FMLP or PMA; a calmodulin-dependent component may rather balance responses induced by IpOCOC9.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Isomerases de Aminoácido/antagonistas & inibidores , Basófilos/metabolismo , Calmodulina/antagonistas & inibidores , Proteínas de Transporte/antagonistas & inibidores , Liberação de Histamina/efeitos dos fármacos , Fosfotransferases/antagonistas & inibidores , Benzimidazóis/farmacologia , Calmodulina/fisiologia , Diacilglicerol Quinase , Felodipino/farmacologia , Humanos , Imidazóis/farmacologia , Isoquinolinas/farmacologia , Peptidilprolil Isomerase , Fenotiazinas/farmacologia , Pirimidinonas/farmacologia , Tacrolimo/farmacologia , Tiazóis/farmacologiaRESUMO
The effects of protein kinase inhibitors on regeneration in vitro of adult frog sciatic sensory axons were tested. Regeneration of crush-injured nerves for 8 days in serum-free medium was inhibited by staurosporine (100 nM) and H-7 (100 microM), which are both known to inhibit protein kinase C. With the use of a compartmented culture system it could be shown that H-7 exerted both local (outgrowth region) and central (ganglia) effects, the latter being more pronounced. The local effects could be due to reduction of Schwann cell proliferation by H-7. Immunohistochemistry demonstrated the presence of protein kinase C in neuronal cell bodies but not in axonal processes. Proliferation of Schwann cells was accompanied by increased protein kinase C immunoreactivity at the site of injury. H-7 caused a selective inhibition in the incorporation of radioactive phosphate into one 74 kDa protein of both ganglia and nerve but also a more general decrease in protein labelling. The results show that protein phosphorylations, possibly mediated by protein kinase C, are involved in regeneration-related mechanisms operating at both local and central levels in the adult frog sciatic sensory axons.
Assuntos
Axônios/enzimologia , Neurônios Aferentes/enzimologia , Inibidores de Proteínas Quinases , Nervo Isquiático/enzimologia , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina , Potenciais de Ação , Alcaloides/farmacologia , Animais , Axônios/efeitos dos fármacos , Divisão Celular , Imuno-Histoquímica , Técnicas In Vitro , Isoquinolinas/farmacologia , Regeneração Nervosa , Neurônios Aferentes/efeitos dos fármacos , Ésteres de Forbol/farmacologia , Fosforilação , Piperazinas/farmacologia , Rana temporaria , Células de Schwann/efeitos dos fármacos , Nervo Isquiático/fisiologia , EstaurosporinaRESUMO
To assess possible involvement of protein kinase C (PKC) in human basophil degranulation, the present work compared effects of various purported PKC inhibitors on leukocyte histamine release triggered by different stimuli. The effects recorded varied with the inhibitor and the secretagogue used; moreover, with a given secretagogue, different inhibitors often displayed different activities. Thus, histamine release triggered by the PKC activator 4 beta-phorbol 12-myristate 13-acetate was blocked by K252a, staurosporine and the purported specific PKC inhibitor Ro 31-7549, and reduced by calphostin C, H-7, TMB-8 and W-7 but not affected by polymyxin B; it was augmented by 2.1 microM palmitoyl carnitine. The leukocyte response induced by another putative activator of PKC, 1,2-isopropylidene-3-decanoyl-sn-glycerol, was also enhanced by 2.1 microM palmitoyl carnitine, slightly increased by staurosporine, TMB-8 and W-7 but not affected by calphostin C, H-7, K252a or Ro 31-7549, whereas the hyperosmolar mannitol-induced response was reduced by H-7, calphostin C, TMB-8 and W-7 and slightly augmented by staurosporine. Anti-IgE-induced histamine release was blocked by staurosporine and K252a and reduced by calphostin C, sphingosine, TMB-8 and W-7 but not affected by H-7, polymyxin B or retinal. It was enhanced by Ro 31-7549. In contrast, leukocyte histamine release induced by calcium ionophore A23187 or by ionomycin was blocked by retinal, TMB-8 and W-7 and reduced by calphostin C and palmitoyl carnitine but enhanced by H-7, staurosporine and polymyxin B; K252a and Ro 31-7549 did not affect such responses. Formyl-methionyl-leucyl-phenylalanine-triggered histamine release was barely affected by any agent used. Thus, the specific PKC inhibitor Ro 31-7549 selectively blocked 4 beta-phorbol 12-myristate 13-acetate-triggered leukocyte histamine release. These results imply that examined secretagogues trigger human leukocyte histamine release through partly separate pathways probably involving different kinase activities (PKC isozymes?). Moreover, the distinct effect patterns recorded for most purported PKC inhibitors imply a functional selectivity between these compounds.
Assuntos
Basófilos/metabolismo , Liberação de Histamina/efeitos dos fármacos , Proteína Quinase C/fisiologia , 1-(5-Isoquinolinasulfonil)-2-Metilpiperazina , Alcaloides/farmacologia , Calcimicina/farmacologia , Éteres de Glicerila/farmacologia , Humanos , Indóis/farmacologia , Isoquinolinas/farmacologia , Maleimidas/farmacologia , Manitol/farmacologia , Palmitoilcarnitina/farmacologia , Piperazinas/farmacologia , Polimixina B/farmacologia , Proteína Quinase C/antagonistas & inibidores , Esfingosina/farmacologia , Estaurosporina , Sulfonamidas/farmacologia , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
1. Galanin concentration-dependently blocked human leukocyte histamine release triggered by the calcium ionophores A23187 and ionomycin. Almost complete inhibition of release was recorded at 410 nM whereas 41 nM galanin mediated close to 50% inhibition of responses. 2. Pretreatment of the cells with pertussis toxin did not influence the inhibitory effects of galanin. 3. Leukocyte responses triggered by anti-IgE, the chemotactic peptide formyl-methionyl-leucyl-phenylalanine (FMLP) or 4 beta-phorbol 12-myristate 13-acetate (PMA) were not affected by galanin. 4. Ionophore-induced basophil histamine release, but not responses triggered by anti-IgE, FMLP or PMA, was inhibited when cells were challenged in medium containing high potassium (120mM). 5. We conclude that galanin and a depolarizing medium selectively inhibit ionophore-induced basophil histamine release.
Assuntos
Calcimicina/farmacologia , Liberação de Histamina/efeitos dos fármacos , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Peptídeos/farmacologia , Ésteres de Forbol/farmacologia , Potássio/farmacologia , Basófilos/efeitos dos fármacos , Basófilos/metabolismo , Galanina , Humanos , Imunoglobulina E/imunologia , Técnicas In Vitro , Toxina Pertussis , Fatores de Virulência de Bordetella/farmacologiaRESUMO
Auranofin, an antiarthritic gold compound, modulates a number of chemotactic factor-induced inflammatory responses in human neutrophils. In order to unravel the mechanism involved, the present study investigated the effects of auranofin on early signal transduction events in these cells. Auranofin did not affect the chemotactic peptide (fMetLeuPhe)-induced formation of inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), neither in the presence nor in the absence of extracellular calcium ions. In contrast, there was a progressive inhibition by auranofin on the fMet-Leu-Phe-induced mobilization of intracellular calcium. This demonstrates that auranofin can dissociate the generation of Ins(1,4,5)P3 from the subsequent release of intracellular calcium, perhaps by interfering with the intracellular binding of Ins(1,4,5)P3 to its receptor. In experiments performed in electro-permeabilized cells, however, a relatively high concentration of the drug failed to abolish the specific binding of Ins(1,4,5)P3. In addition, in the same system, auranofin also failed to abolish the Ins(1,4,5)P3-induced release of Ca2+. Consequently, auranofin-mediated dissociation of fMLP-induced Ins(1,4,5)P3 formation and intracellular calcium release can not be explained merely by an antagonistic effect of auranofin on the Ins(1,4,5)P3 receptor. Instead the interaction between auranofin and the plasma membrane seems to be an initial and important part of the mechanism by which this drug interferes with the transduction signalling system.
Assuntos
Auranofina/farmacologia , Canais de Cálcio , Cálcio/sangue , Inositol 1,4,5-Trifosfato/sangue , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Neutrófilos/metabolismo , Receptores Citoplasmáticos e Nucleares , Aminoquinolinas , Ácido Egtázico/farmacologia , Corantes Fluorescentes , Humanos , Técnicas In Vitro , Inositol 1,4,5-Trifosfato/biossíntese , Receptores de Inositol 1,4,5-Trifosfato , Cinética , Neutrófilos/efeitos dos fármacos , Receptores de Superfície Celular/efeitos dos fármacos , Receptores de Superfície Celular/metabolismoRESUMO
Selected functions of alveolar macrophages obtained by bronchoalveolar lavage of 12 healthy smokers were examined before and after eight weeks' treatment with an inhaled glucocorticosteroid, budesonide (400 micrograms twice daily). After budesonide treatment spontaneous as well as opsonised zymosan triggered prostaglandin E2 (PGE2) secretion from harvested cells was reduced; no such reduction in opsonised zymosan triggered leukotriene B4 (LTB4) production was observed. Neither the capacity to phagocytose opsonised yeast particles nor the superoxide radical generation triggered by the calcium ionophore A23187, 4 beta-phorbol 12-myristate 13-acetate (PMA), or opsonised zymosan ex vivo were more than marginally affected by the glucocorticosteroid treatment in vivo. Lavage fluid concentrations of angiotensin converting enzyme (ACE), however, after treatment were twice those before treatment and concentrations of fibronectin were reduced to half. Albumin concentrations in lavage fluid were not affected by the glucocorticosteroid treatment. In separate experiments treatment of alveolar macrophages with 10(-7) or 10(-6) M budesonide overnight in vitro did not affect their superoxide radical or PGE2 generation but significantly blocked LTB4 release. These data indicate that inhaled gluco-corticosteroid treatment may affect synthesis or release (or both) of ACE and fibronectin by alveolar macrophages from healthy smokers whereas other functions of these cells, such as the generation of reactive oxygen derived products ex vivo, are only marginally affected.
Assuntos
Glucocorticoides/administração & dosagem , Macrófagos/efeitos dos fármacos , Pregnenodionas/administração & dosagem , Alvéolos Pulmonares/efeitos dos fármacos , Fumar/metabolismo , Administração por Inalação , Adulto , Líquido da Lavagem Broncoalveolar/análise , Budesonida , Dinoprostona/biossíntese , Feminino , Fibronectinas/análise , Radicais Livres , Humanos , Leucotrieno B4/biossíntese , Macrófagos/fisiologia , Masculino , Peptidil Dipeptidase A/análise , Fagocitose/efeitos dos fármacos , SuperóxidosRESUMO
The generation of reactive oxygen-derived species is one main constituent of the microbicidal activity of professional phagocytes. This process is known as the respiratory or the oxidative burst. It is initiated by a cyanide- and azide-insensitive increase in O2-consumption and the concomitant generation of superoxide radicals catalyzed by a membrane-localized NADPH oxidase which is triggered by an appropriate stimulation of the cells. The generated O2 is converted to hydrogen peroxide, hydroxyl radicals and other reactive products of oxygen which, if released extracellularly (for example in connection with frustrated phagocytosis), are potentially harmful to the tissue. The oxidative burst is not necessarily dependent on phagocytosis, nor is it necessarily associated with degranulation. Therefore, the process constitutes an important independent variable of phagocyte activity, and researches aiming to characterize various forms of airway inflammation may derive valuable information from an examination of the oxidative burst.
Assuntos
Oxigênio/sangue , Fagócitos/metabolismo , HumanosRESUMO
1. Diadenosine tetraphosphate (Ap4A) levels were determined in HL-60 cells differentiating into granulocytes or monocytes after treatment for 0-7 days with retinoic acid (RA) or 4-beta-phorbol-12-myristate-13-acetate (PMA) respectively. 2. The levels increased significantly compared to untreated control cells within 2 days and then declined again. 3. In RA treated cells the levels finally decreased far below those of untreated HL-60 cells and became equal to those found in human granulocytes. 4. PMA treatment had no effect on Ap4A levels in human granulocytes. 5. A possible interaction between Ap4A and ADP-ribosyl transferase is discussed.
Assuntos
Fosfatos de Dinucleosídeos/metabolismo , Granulócitos/citologia , Leucemia Promielocítica Aguda/metabolismo , Monócitos/citologia , Diferenciação Celular , Granulócitos/efeitos dos fármacos , Granulócitos/metabolismo , Humanos , Leucemia Promielocítica Aguda/patologia , Acetato de Tetradecanoilforbol/farmacologia , Tretinoína/farmacologia , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/metabolismoRESUMO
The mechanical responses of circular segments of uterine arteries to different combinations of vasoactive agents and putative inhibitors of calcium fluxes were examined using a sensitive in vitro method. Exposure to high potassium (127 mM) or to noradrenaline (NA; 0.1 mM) resulted in a rapid, initial increase in tension of the vascular preparation to reach a sustained level of contraction (approximatively 12 mN) which lasted for at least 15 min. The protein kinase C activator, 4-beta-phorbol-12,13-esther-dibutyrate (10 microM), induced a slowly developing but sustained contractile response with a maximum (PDBumax) of only 4 mN. Addition of the calcium ionophore, A23187, to PDBu-contracted vessels did not increase tension, while the maximum tension evoked by A23187 per se (3 microM) was 5 mN. Ionomycin had only a small contractile effect on the uterine artery. The contraction evoked by depolarization (potassium) or alpha 1-adrenoceptor stimulation (NA) was decreased in nominally calcium-free medium containing EDTA (0-100 microM) or EGTA, while uterine arteries loaded intracellularly with the calcium chelator, quin-2, responded to the vasoconstrictors almost as well as the control preparations. Blockade of calcium influx with Cd2+ (greater than 0.01 mM), nifedipine (greater than 3 microM), verapamil (greater than 1 microM) and TMB-8 (greater than 10 microM) reduced the tension evoked by potassium somewhat more than it reduced the contractions induced by NA, while the opposite was seen in the presence of Ni2+ (greater than 0.1 mM). Inhibition of calmodulin-dependent enzymes by W7 (greater than 10 microM) reduced the maximum tension evoked by potassium and NA to a similar extent.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Cálcio/farmacologia , Proteína Quinase C/metabolismo , Útero/irrigação sanguínea , Vasoconstrição/efeitos dos fármacos , Animais , Bloqueadores dos Canais de Cálcio/farmacologia , Ativação Enzimática/efeitos dos fármacos , Feminino , Cobaias , Técnicas In Vitro , Contração Muscular/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Norepinefrina/farmacologia , Fluxo Sanguíneo Regional/efeitos dos fármacos , Frações Subcelulares/efeitos dos fármacosRESUMO
Compound 1,2-isopropylidene-3-decanoyl-sn-glycerol (IpOCOC9) augments the phosphorylation in vitro of histone III-S and myelin basic protein (MBP) by a partially purified Ca2(+)- and phospholipid-dependent protein kinase activity (protein kinase C) from human polymorphonuclear leukocytes. IpOCOC9 can substitute for either Ca2+ and phosphatidylserine or for phorbol ester. The related compound decanoid acid cyclopentyl methylester (DACPME) is less effective than IpOCOC9 in this respect. These data lend support to the notion that the secretagogue activity of IpOCOC9 with respect to human basophil histamine release and neutrophil superoxide radical generation is due to protein kinase C activation.
Assuntos
Éteres de Glicerila/farmacologia , Neutrófilos/enzimologia , Proteína Quinase C/metabolismo , Cálcio/metabolismo , Ciclopentanos/farmacologia , Ácidos Decanoicos/farmacologia , Ativação Enzimática , Histamina/metabolismo , Histonas/metabolismo , Humanos , Hidroxiapatitas , Proteína Básica da Mielina/metabolismo , Ésteres de Forbol/metabolismo , Fosfatidilserinas/metabolismo , FosforilaçãoRESUMO
Previous studies have shown that the glyceride derivative, sn-1,2-isopropylidene-3-decanoyl-glycerol (IpOCOC9), can trigger human leukocyte histamine release. Approximately 25% of the total cellular histamine content is extruded in the presence of 206 microM of IpOCOC9; at 69 microM, however, the secretagogue action of the compound is marginal. The characteristics of the release induced by IpOCOC9 are closely similar to those reportedly recorded at hyperosmolar triggering of basophils with mannitol, and in many respects they also mimic those observed at phorbol ester-induced histamine release. The compound decanoic acid cyclopentyl methylester (DACPME), a structural analogue of IpOCOC9, fails to induce histamine release. IpOCOC9, but not DACPME, stimulates human polymorphonuclear leukocyte cytosolic Ca2+- and phospholipid-dependent histone III-S kinase activity (unpublished observations). The secretagogue action of IpOCOC9 has therefore tentatively, at least partly, been attributed to a direct protein kinase C activation. In the present studies, we examined the influence of IpOCOC9 and DACPME on histamine release triggered by an ensuing exposure to anti-IgE, the calcium ionophore A23187, formyl-methionyl-leucyl-phenylalanine (FMLP), or 4 beta-phorbol 12-myristate 13-acetate (PMA). It is shown that IpOCOC9-treatment of cells results in either enhancement or reduction of the release induced by anti-IgE or by A23187, whereas FMLP-induced release is consistently reduced and PMA-induced release consistently enhanced by such a treatment. Treatment of cells with DACPME enhances but does not reduce anti-IgE-triggered release, whereas FMLP-induced release is not affected. Pretreatment of the cells with other putative protein kinase C activators like PMA, sn-1-oleoyl-2-acetyl-glycerol (OAG), 1,2-dioctanoyl-glycerol (DiC8) or the glycerol derivative sn-1,2-diacetyl-3-decanoyl-glycerol (DiC2OCOC9) affects secretagogue-induced basophil histamine release according to specific patterns similar to but not identical with those recorded for IpOCOC9 and DACPME. Thus, e.g., DiC2OCOC9 consistently reduces but does not enhance anti-IgE-triggered release. These data show that limited structural changes of IpOCOC9 may qualitatively affect its modulating properties in the human basophil histamine release system.
Assuntos
Ciclopentanos/farmacologia , Ácidos Decanoicos/farmacologia , Éteres de Glicerila/farmacologia , Liberação de Histamina/efeitos dos fármacos , Leucócitos/metabolismo , Calcimicina/farmacologia , Diglicerídeos/farmacologia , Humanos , Imunoglobulina E/antagonistas & inibidores , Leucócitos/efeitos dos fármacos , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Acetato de Tetradecanoilforbol/análogos & derivados , Acetato de Tetradecanoilforbol/farmacologia , Triglicerídeos/farmacologiaRESUMO
Human leukocytes were found to release histamine at exposure for the synthetic glyceride derivative sn-1,2-isopropylidene-3-decanoyl-glycerol (IpOCOC9). The following characteristics for the IpOCOC9-induced basophil histamine release were recorded. A. In the order of 25% of the cellular histamine content was extruded at 206 mumol/l and 45% at 690 mumol/l of the compound, respectively. B. Removal of extracellular Ca2+ variably affected IpOCOC9-triggered release. C. The presence of N-ethylmaleimide (10 mumol/l) or p-bromophenacylbromide (10 mumol/l) markedly reduced IpOCOC9-induced histamine release. D. The time course of the release triggered by IpOCOC9 was intermediate to those characterizing the release triggered by 4 beta-phorbol 12-myristate 13-acetate (PMA) and by formyl-methionyl-leucyl-phenylalanine (FMLP). E. Cells desensitized to IgE-receptor-mediated stimulation were hyperresponsive to stimulation with IpOCOC9. F. Cells treated with a low concentration of 2-deoxyglucose were not hyperresponsive to IpOCOC9. These data show that IpOCOC9, a PMN/leukocyte protein kinase C stimulator, acts as a non-cytotoxic secretagogue for human basophils with a mode of action which in some, but not all respects, mimics that of PMA. In particular, IpOCOC9-triggered release resembles that reported by other authors for hyperosmolar triggering of release by mannitol.
Assuntos
Basófilos/efeitos dos fármacos , Ácidos Decanoicos/farmacologia , Dioxóis/farmacologia , Liberação de Histamina/efeitos dos fármacos , Proteína Quinase C/metabolismo , Triglicerídeos/farmacologia , Acetofenonas/farmacologia , Antígenos de Diferenciação de Linfócitos B/fisiologia , Basófilos/metabolismo , Cálcio/metabolismo , Ciclopentanos/farmacologia , Ácidos Decanoicos/antagonistas & inibidores , Desoxiglucose/farmacologia , Ativação Enzimática/efeitos dos fármacos , Etilmaleimida/farmacologia , Temperatura Alta , Humanos , N-Formilmetionina Leucil-Fenilalanina/farmacologia , Receptores Fc/fisiologia , Receptores de IgE , Taxa Secretória/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Estimulação Química , Acetato de Tetradecanoilforbol/farmacologia , Triglicerídeos/antagonistas & inibidoresRESUMO
Bronchoalveolar lavage (BAL) was performed on eleven healthy smokers before and after eight weeks of oral treatment with N-acetylcysteine (NAC) 200 mg t.i.d. The concentrations of selected eosinophil and neutrophil granule constituents and of selected proteases and protease inhibitors, albumin and endotoxin were determined in the recovered BAL fluid and in plasma or serum samples. In addition, in vitro chemotactic activities for neutrophils and eosinophils were assessed in the BAL fluid. Significant reductions in BAL fluid content of lactoferrin (LF), eosinophil cationic protein (ECP), antichymotrypsin (ACT) and chemotactic activity for neutrophils were recorded after NAC treatment. The levels of other examined markers tended to be reduced or were not affected. In serum/plasma, the concentrations of myeloperoxidase (MPO) and elastase were reduced after NAC treatment whereas concentrations of other constituents examined were unaltered. These data, together with previously reported findings, suggest that oral NAC may influence the activity of "inflammatory" cells in the bronchoalveolar space of smokers.
Assuntos
Acetilcisteína/administração & dosagem , Biomarcadores/análise , Líquido da Lavagem Broncoalveolar/citologia , Ribonucleases , Fumar/metabolismo , Administração Oral , Adulto , Proteínas Sanguíneas/análise , Endotoxinas/análise , Proteínas Granulares de Eosinófilos , Humanos , Lactoferrina/análise , Masculino , Muramidase/análise , Elastase Pancreática/análise , Peroxidase/análise , Inibidores de Proteases/análise , Albumina Sérica/análiseRESUMO
IgE-receptor dependent, but not A23187-induced, histamine release from passively sensitized chopped human lung tissue, or from a dispersed lung cell population obtained from the tissue after enzymatic digestion, is reduced after incubation overnight of cells/tissue with the glucocorticosteroid budesonide (10(-7) M). Since the inhibitory effect of budesonide is reduced in the continuous presence of diluted reagin-rich serum used for passive sensitization, but not in the presence of heat-inactivated serum, it is suggested that the glucocorticosteroid acts by reducing the binding of IgE-antibody to the target cell(s).
Assuntos
Liberação de Histamina/efeitos dos fármacos , Pulmão/metabolismo , Pregnenodionas/farmacologia , Anticorpos Anti-Idiotípicos/imunologia , Budesonida , Calcimicina/farmacologia , Células Cultivadas , Glucocorticoides/farmacologia , Humanos , Imunoglobulina E/imunologia , Pulmão/citologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Pólen/imunologia , Reaginas/imunologiaRESUMO
We examined the effects of glucocorticosteroids (GCS) on antigen-induced bronchial anaphylactic reactions (BAR) in SD rats immunized with ovalbumin (OA) and alum. The animals were treated with vehicle, budesonide (BUD), dexamethasone (DEX), or hydrocortisone (HC) at various times before intravenous (i.v.) antigen challenge. The drugs were administered either intraperitoneally (i.p.) or intratracheally (i.t.); the BAR was elicited by a low or by a high challenge dose of antigen. A BAR elicited by a low challenge dose of antigen was reduced in a dose-dependent way by all GCS after i.p. administration; at 1 mg/kg, BUD and DEX significantly reduced BAR and at 50 mg/kg all three of the examined compounds inhibited the BAR by 50% or more. For BUD, maximum effect was recorded when it was given 12 h before test. There was only a slight variation in the inhibitory effects of the GCS with immunization conditions of test animals. I.t. instillation of the drugs did not markedly increase their inhibitory capacity as compared to i.p. administration. BAR elicited by a high antigen dose was at best marginally affected by the GCS when given either i.p. or i.t. Thus, antigen-induced airway reactivity in rats can be reduced by GCS treatment provided that this is performed sufficiently long before the test and that the challenge dose of antigen is not too high.
Assuntos
Compostos de Alúmen , Anafilaxia/imunologia , Antígenos/farmacologia , Broncopatias/imunologia , Glucocorticoides/farmacologia , Respiração/efeitos dos fármacos , Acetofenonas/farmacologia , Administração Tópica , Alumínio/imunologia , Anafilaxia/sangue , Anafilaxia/tratamento farmacológico , Animais , Budesonida , Cromolina Sódica/farmacologia , Dexametasona/administração & dosagem , Dexametasona/farmacologia , Relação Dose-Resposta a Droga , Interações Medicamentosas , Glucocorticoides/administração & dosagem , Imunização , Injeções Intraperitoneais , Injeções Intravenosas , Masculino , Ovalbumina/imunologia , Pregnenodionas/administração & dosagem , Pregnenodionas/farmacologia , Quinacrina/farmacologia , Ratos , Ratos Endogâmicos/imunologia , Sulfatos/imunologia , Fatores de TempoRESUMO
Bronchoalveolar lavage (BAL) was performed on nine healthy nonsmoking subjects and on 11 healthy smokers; in the last mentioned group lavage was performed before and after eight weeks treatment with N-acetylcysteine (NAC; 200 mg t.i.d.). The BAL cells were cultured for 2 h or overnight. Adherent cells were examined for their capacity to generate superoxide radicals (determined by superoxide dismutase (SOD)-inhibitable cytochrome C-reduction) at stimulation with phorbol 12-myristate 13-acetate (PMA), serum-treated zymosan (STZ), the calcium ionophore A23187, or the chemotactic tripeptide formyl-methionylleucylphenylalanine (FMLP). Cells from nonsmokers responded with a very low degree of O(2)-generation to any of the stimuli employed whether cultured for 2 h or overnight. Cells from smokers also responded with low O(2)-generation after 2 h of culture. However, cells from smokers cultured overnight responded with marked O(2)-generation to PMA and STZ but the responses to FMLP and A23187 were low. NAC-treatment of the smokers resulted in a reduced degree of both PMA- and STZ-induced O(2)-generation in five individuals. In two other subjects, PMA-induced (but not STZ-induced) O(2)-generation was reduced. Two individuals showed increased O(2)-generation to PMA- and to STZ-stimulation after NAC-treatment. Mean values of O(2)-generation induced by A23187 or by FMLP were significantly reduced for cells harvested after NAC-treatment. Mean values for PMA-induced O(2)-generation also tended to be reduced by the treatment.(ABSTRACT TRUNCATED AT 250 WORDS)