RESUMO
Studies were made on the ability of the leukemic cell line, HL-60, to substitute for normal human granulocytes in research concerned with the bioactivation of arylamines. The arylamine carcinogen, 2-aminofluorene (2-AF), was used as the model substrate in the form of 2-[9-14C]AF, and was incubated with HL-60 cell cultures, both in the presence and absence of phorbol myristate acetate (PMA) which induces the respiratory burst. The HL-60 cultures were generally employed after having been induced to undergo differentiation to neutrophils by the action of dimethyl sulfoxide (DMSO). Comparisons of the amounts of DNA and RNA binding by 2-AF between HL-60 and normal human granulocyte cultures demonstrated close similarities in the amount and nature of nucleic acid binding by this arylamine substrate. HL-60 cells that had been induced to differentiate to neutrophils to the extent of about 80% showed high levels of the respiratory burst along with extensive covalent binding of 2-[9-14C]AF to cellular nucleic acids. Although normal human granulocytes tended to metabolize 2-AF slightly faster than did highly differentiated HL-60 cells, the extent of nucleic acid binding relative to the amount of 2-AF metabolized was similar. A major difference in the metabolic fate of 2-AF in these cell cultures was the unique ability of HL-60 cultures at all stages of differentiation to effect the slow N-acetylation of 2-AF to give 2-acetylaminofluorene (2-AAF). Extensive analyses of incubation extracts showed that the major differences in apparent metabolites were quantitative. With few exceptions, both activated HL-60 and granulocyte cell cultures produced the same metabolites, most of which remain unidentified. Studies with inhibitors such as catalase, superoxide dismutase and azide ion further suggest that these two related cell cultures metabolize 2-AF in similar manner. The DMSO-differentiated HL-60 culture is proposed as a convenient model with which to investigate the metabolism and bioactivation of arylamines by human granulocytes or pure neutrophils.
Assuntos
Aminas/metabolismo , Carcinógenos/metabolismo , DNA/metabolismo , Fluorenos/metabolismo , Granulócitos/metabolismo , Leucemia Experimental/metabolismo , Leucemia Mieloide/metabolismo , RNA/metabolismo , Xenobióticos/metabolismo , Aminas/sangue , Diferenciação Celular , Cromatografia Líquida de Alta Pressão/métodos , Dimetil Sulfóxido/farmacologia , Humanos , Leucemia Experimental/patologia , Leucemia Mieloide/patologia , Neutrófilos/citologia , Neutrófilos/metabolismo , Oxigênio/metabolismo , Consumo de Oxigênio , Ligação Proteica , Acetato de Tetradecanoilforbol/farmacologia , Células Tumorais Cultivadas , Xenobióticos/sangueRESUMO
The aim of this study was to investigate the oxidative effects of Fe(2)O(3), benzo(a)pyrene (B(a)P) and pyrene, alone or in association (B(a)P or pyrene coated onto Fe(2)O(3) particles), in normal human embryonic lung epithelial cells (L132) in culture. We evaluated: (i) membrane integrity, through fatty acid release (stearic acid, oleic acid, linoleic and linolenic acids, homolinolenic acid, arachidonic acid) and malondialdehyde (MDA) production; and (ii) antioxidant status, through enzymatic and non-enzymatic antioxidant defenses (superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), glutathione status, beta-carotene). Fe(2)O(3) did not induce any change in L132 cells. In pyrene-treated cells, SOD induction (P<0. 05), glutathione oxidation (P<0.05) and beta-carotene consumption (P<0.001) may counteract free radicals (FR)-induced damage. However, in B(a)P-incubated cells, SOD inactivation (P<0.05), GR increases (P<0.05), glutathione oxidation (P<0.05) and beta-carotene decreases (P<0.001) showed high disruption of antioxidants, thereby allowing FR-induced damage (i.e. arachidonic acid release, P<0.01; MDA production, P<0.01). Our main finding was that both associations caused higher FR-induced damage (i.e. MDA production, P<0.001; SOD inactivation, P<0.01) than either chemical alone. Several mechanisms could account for this result: enhanced uptake of Fe(2)O(3) particles and/or greater availability of polycyclic aromatic hydrocarbons (PAHs). We hypothesized also that Fe(2)O(3) and polycyclic aromatic hydrocarbons are more deleterious by virtue of their associations being able to produce higher oxidative effects than either chemical alone.
Assuntos
Compostos Férricos/farmacologia , Hidrocarbonetos Policíclicos Aromáticos/farmacologia , Antioxidantes/metabolismo , Benzo(a)pireno/farmacologia , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Células Epiteliais/citologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Ácidos Graxos/metabolismo , Glutationa/efeitos dos fármacos , Glutationa/metabolismo , Glutationa Peroxidase/efeitos dos fármacos , Glutationa Peroxidase/metabolismo , Glutationa Redutase/efeitos dos fármacos , Glutationa Redutase/metabolismo , Humanos , Pulmão/citologia , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Malondialdeído/metabolismo , Tamanho da Partícula , Pirenos/farmacologia , Superóxido Dismutase/efeitos dos fármacos , Superóxido Dismutase/metabolismoRESUMO
Epidemiological evidence firmly implicated an interactive effect between Fe2O3 and benzo(a)pyrene (B(a)P) in causing lung cancer. However, despite intensive investigation, the mechanism involved is not precisely established. Since the accumulation of reactive oxygen intermediates (ROI)-mediated damage and/or immune-induced injury might be a possible cause of lung cancer, we studied the oxidative and the inflammatory effects of Fe2O3 (3 mg), B(a)P (3 mg) or B(a)P (3 mg)-coated onto Fe2O3 (3 mg) particles on this relevant organ target in Sprague-Dawley rats. We investigated lipid peroxidation (malondialdehyde; MDA) and secretion of some inflammatory mediators (tumor necrosis factor-alpha, TNF-alpha; interleukin-1 beta, IL-1beta; nitric oxide, NO) in lungs. In addition, mRNA expressions of TNF-alpha, IL-1beta and inducible nitric oxide synthase (iNOS) were evaluated. Our results show that exposure to Fe2O3 and B(a)P, alone or in association, induced 2-fold increases in MDA production suggesting thereby oxidative stress conditions (P<0.01). Exposure to Fe2O3, B(a)P or B(a)P-coated onto Fe2O3 particles significantly increased both mRNA expression and/or synthesis of inflammatory mediators. The main findings of this work were that the association of Fe2O3 and B(a)P induces more pronounced induction of inflammatory mediators (IL-1beta secretion, P<0.01; IL-1beta mRNA expression, P<0.01; iNOS mRNA expression, P<0.05) than B(a)P by itself. Hence, our results may explain why concurrent exposure to Fe2O3 and B(a)P is more deleterious in lungs than exposure to B(a)P alone.
Assuntos
Benzo(a)pireno/toxicidade , Compostos Férricos , Pulmão/efeitos dos fármacos , Pneumonia/induzido quimicamente , Animais , Benzo(a)pireno/administração & dosagem , Líquido da Lavagem Broncoalveolar/química , Meios de Cultura/análise , Interações Medicamentosas , Interleucina-1/análise , Interleucina-1/imunologia , Pulmão/imunologia , Pulmão/metabolismo , Macrófagos Alveolares/metabolismo , Masculino , Malondialdeído/análise , Malondialdeído/metabolismo , Óxido Nítrico/análise , Óxido Nítrico Sintase/análise , Óxido Nítrico Sintase/biossíntese , Óxido Nítrico Sintase Tipo II , Pneumonia/imunologia , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Organismos Livres de Patógenos Específicos , Traqueia/efeitos dos fármacos , Traqueia/imunologia , Traqueia/metabolismo , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/imunologiaRESUMO
In Senegal, as in many developing countries, traffic density is increasing in urban areas; in Dakar more than 50% of vehicles use gasoline. Yet the extent and real magnitude of the problem has neither been recognized nor assessed in these countries. Systemic data assessment of lead pollution and people's exposure are not well known in Senegal. This study was also designed to determine the impregnation levels of the lead released by the exhaust of cars and the changes of some early biological markers in Senegalese children. Blood lead (BPb) levels showed that all the children enrolled were exposed. However, lead exposure levels (from 34.7 to 145.8 microg/L) were less important for children living in rural areas (60.9+/-18.3 microg/L) than for those living in urban areas (106.7+/-16.9 microg/L). These changes could be correlated to the difference in the automobile traffic between both these regions (P < 0.001). BPb mean levels found in boys were higher than those in girls (P < 0.05). Despite elevated BPb levels, all values for blood zinc protoporphyrin and urine delta-aminolevulinic acid were within physiological ranges. In addition, variations in some biological markers of oxidative stress and renal disorders were seen; however, they must be confirmed by a future epidemiological study.
Assuntos
Poluentes Atmosféricos/sangue , Chumbo/sangue , Estresse Oxidativo/efeitos dos fármacos , Emissões de Veículos/efeitos adversos , Poluentes Atmosféricos/efeitos adversos , Criança , Feminino , Humanos , Chumbo/efeitos adversos , Masculino , Projetos Piloto , População Rural , Senegal , População UrbanaRESUMO
A method has been developed for the determination of acetaminophen in human serum. Samples are extracted with ethyl acetate. Beta-hydroxyethyl theophylline is used as internal standard. The solvent is evaporated under nitrogen and the residue is analyzed by high-performance liquid chromatography on Lichrosorb RP 18. Extraction efficiency, linearity and assay precision have been determined. The method is rapid and can be easily applied to determine the plasma level of patient having taken overdose of paracetamol.
Assuntos
Acetaminofen/sangue , Cromatografia Líquida de Alta Pressão/métodos , HumanosRESUMO
Iron oxides are present in many occupational atmospheres mainly in iron ore mines and in steel industry. Among these workers, epidemiological studies indicated an excess of lung cancer deaths. In mines, it was difficult to involve iron oxides exposure because there are other possible causes as radon, polycyclic aromatic hydrocarbon (PAH) present in diesel exhausts, silicosis or siderosis. The contradictory results of these studies are due to the differences of exposure levels or to the presence or not of these cofactors or of a sufficient prevention. But generally the results agree with an interaction of iron oxide dusts and smoking habits. It is unclear if this interaction supports an additive or multiplicative risk of lung cancer. Experimental studies with Fe2O3 showed that these particles are able to induce lung cancers only in the presence of PAH when administered to animals. In vitro studies permitted to observe an interaction in the metabolism of benzo(a)pyrene (BaP) leading to a higher level of precursors of the ultimate carcinogen. As this metabolism of BaP is known to be enhanced during lipoperoxidation, it is possible to involve this mechanism with Fe2O3. After phagocytosis and dissolution with production of ferric ions, Fe2O3 can enhance the production of reactive oxygen species responsible of damaging both lipidic constituents and DNA. Fe3O4 and mainly FeO may be more toxic, introducing directly ferrous ions in the cells after dissolution, but the cancerogenicity of the these compounds is unknown, making necessary to develop research.
Assuntos
Compostos Férricos/metabolismo , Neoplasias Pulmonares/induzido quimicamente , Mineração , Doenças Profissionais/induzido quimicamente , Exposição Ocupacional , Hidrocarbonetos Policíclicos Aromáticos , Animais , Benzo(a)pireno/metabolismo , Compostos Férricos/efeitos adversos , Compostos Ferrosos/metabolismo , Humanos , Peroxidação de Lipídeos , Neoplasias Pulmonares/epidemiologia , Doenças Profissionais/epidemiologia , Estresse Oxidativo , Hidrocarbonetos Policíclicos Aromáticos/efeitos adversos , Hidrocarbonetos Policíclicos Aromáticos/metabolismoRESUMO
Simple improvements of the crossed immunoelectrophoresis method are described. A trap-gel was prepared with a small quantity of monospecific antiserum and was submitted to preelectrophoresis. It blocked, during the first dimension, an individual protein as a rocket. The corresponding peak was reduced or disappeared in the second dimension. Identification of precipitation peaks in human or Macacus cynomolgus bronchoalveolar lavage fluids is illustrated and unequivocal recognition of some antigens is shown.
Assuntos
Imunoeletroforese Bidimensional/métodos , Imunoeletroforese/métodos , Proteínas/isolamento & purificação , Animais , Antitrombina III/isolamento & purificação , Brônquios/imunologia , Reações Cruzadas , Humanos , Imunoglobulina A/isolamento & purificação , Macaca fascicularis , Alvéolos Pulmonares/imunologia , Irrigação Terapêutica , Transferrina/isolamento & purificação , Proteína de Ligação a Vitamina D/isolamento & purificação , alfa-Macroglobulinas/isolamento & purificaçãoRESUMO
After being induced to differentiate into a neutrophilic type, cultures of the leukemic cell line HL-60 were able to cause the bioactivation and nucleic acid binding of acetaminophen upon stimulation of the respiratory burst. This phenomenon was found to simulate the same process as that previously shown with normal human granulocytes. Binding to both DNA and RNA of the cells was determined quantitatively by use of 14C-labeled acetaminophen congeners. Protein binding occurred to about the same extent as did RNA binding. Simultaneous labeling experiments with [ring-14C]- and [14C = O]acetaminophen further showed that the acetaminophen molecule was bound to DNA in an intact manner, while binding to RNA showed about a 50% excess binding of the acetaminophen ring relative to the carbonyl group. Experiments with certain inhibitors showed that catalase and azide ion strongly inhibited DNA binding, while superoxide dismutase had a slight stimulatory effect on binding. These results suggest a significant role for myeloperoxidase in the bioactivation process, which contrasts with the proposed bioactivation mechanism of certain arylamine compounds. A mechanism was proposed for acetaminophen binding to nucleic acids that requires the 1 e- oxidation of this substrate to its phenoxyl radical, although the production of the N-acetyl-p-benzoquinoneimine metabolite, which has been proposed to account for the extensive protein binding known to occur for acetaminophen, might also contribute to such binding. The potential genotoxicity of acetaminophen was considered in view of what might be a unique pathway which can metabolize this chemical to a nucleic acid-binding species.
Assuntos
Acetaminofen/metabolismo , DNA de Neoplasias/metabolismo , RNA Neoplásico/metabolismo , Linhagem Celular , DNA de Neoplasias/isolamento & purificação , Dimetil Sulfóxido/farmacologia , Granulócitos/metabolismo , Humanos , Leucemia/metabolismo , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , RNA Neoplásico/isolamento & purificação , Explosão RespiratóriaRESUMO
1. Serial bronchoalveolar lavages were performed on a subhuman primate (Macacus cynomolgus) in order to give an experimental model for silicosis. 2. We have measured glycosidases, proteases, peroxidase and antiproteases of the BAL fluids from seven normal monkeys. 3. The results obtained were similar to those found in human control BAL fluids. 4. For monkeys, the repetition of the bronchoalveolar procedure does not seem to have an important influence on the values obtained. 5. The present results will now permit sequential follow up studies during the course of experimental silicosis.
Assuntos
Líquido da Lavagem Broncoalveolar/enzimologia , Hidrolases/metabolismo , Peptídeo Hidrolases/metabolismo , Peroxidases/metabolismo , Inibidores de Proteases/análise , Animais , Líquido da Lavagem Broncoalveolar/análise , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Imunoeletroforese Bidimensional , Macaca fascicularisRESUMO
Following stimulation with phorbol myristate acetate, human granulocytes were found to incorporate acetaminophen, p-phenetidine, p-aminophenol, and p-chloroaniline into cellular DNA and RNA. Phenacetin was not incorporated into nucleic acid or metabolized by such activated granulocytes. None of the substrates gave nucleic acid binding if the granulocyte cultures were not induced to undergo the respiratory burst. Additional studies on the binding of acetaminophen to DNA and RNA were made by use of both ring-14C-labeled and carbonyl-14C-labeled forms of this substrate. The finding that equivalent amounts of these two labeled acetaminophen substrates were bound to cellular DNA demonstrated that the intact acetaminophen molecule was incorporated into DNA. On the other hand, the finding that excess ring-14C-labeled acetaminophen was incorporated into cellular RNA implies partial hydrolysis of the acetaminophen substrate prior to RNA binding. Evidence was presented which strongly indicates that the nucleic acid binding of the substrates was covalent in nature. The inability of the respiratory burst to result in the binding of phenacetin to nucleic acid suggests that arylamides are not normally activated or metabolized by activated granulocytes. Acetaminophen is an exception to the recalcitrance of arylamides to such bioactivation processes because it also possesses the phenolic functional group, which, like the arylamine group, is oxidized by certain reactive oxygen species. Myeloperoxidase appears to be much more important in the binding of acetaminophen to DNA than it is in the DNA binding of arylamines in general. The role of the respiratory burst in causing the bioactivation of certain arylamines, which are not normally genotoxic via the more usual microsomal activation pathways, was extended to include certain amide substrates such as acetaminophen.
Assuntos
Acetaminofen/metabolismo , DNA/metabolismo , Granulócitos/metabolismo , RNA/metabolismo , Explosão Respiratória , Acetaminofen/farmacocinética , Aminas/metabolismo , Azidas/farmacologia , Biotransformação , Cromatografia Líquida de Alta Pressão , DNA/efeitos dos fármacos , Diálise , Humanos , Técnicas In Vitro , Oxirredução , Peroxidase/antagonistas & inibidores , RNA/efeitos dos fármacos , Azida Sódica , Solubilidade , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
Pneumoconiosis is defined as the disease resulting from a chronic exposure to different inorganic dusts. In order to assess the lung defence against the effects of dust exposure, we studied the bronchoalveolar lavage (BAL) fluids from 30 silicotic patients (9 of them having a diagnosis of progressive massive fibrosis (PMF)) and 8 subjects with a diagnosis of asbestosis. Total protein content, N-acetyl-beta-D-glucosaminidase activity, free elastase-like activity, immunoreactive alpha 1-proteinase inhibitor (alpha 1PI) and neutrophil elastase inhibitory capacity (NEIC) were determined, and the values obtained were compared to those of 14 control BAL fluids. In all of the patients, our data showed a significant increase of total protein content and free elastase-like activity. In contrast, N-acetyl-beta-D-glucosaminidase activities did not reach statistical significance. Values concerning immunoreactive alpha 1PI and NEIC were significantly raised only in patients with PMF and with asbestosis. When the ratio NEIC/immunoreactive alpha 1PI was calculated, a significant difference was noticed in the asbestosis group; on the other hand, this ratio was significantly reduced in the group of PMF patients. After neutrophil elastase addition, an electrophoretic study by SDS-PAGE and immunoblotting was carried out; it showed more proteolysed alpha 1PI in the BAL fluids having a lowered NEIC/alpha 1PI ratio. These facts could be explained by the presence of inhibitors of neutrophil elastase different from alpha 1PI.
Assuntos
Asbestose/metabolismo , Líquido da Lavagem Broncoalveolar/metabolismo , Elastase Pancreática/antagonistas & inibidores , Silicose/metabolismo , Acetilglucosaminidase/metabolismo , Contagem de Células Sanguíneas , Líquido da Lavagem Broncoalveolar/citologia , Eletroforese em Gel de Poliacrilamida , Feminino , Humanos , Immunoblotting , Masculino , Neutrófilos/enzimologia , Elastase Pancreática/metabolismo , alfa 1-Antitripsina/metabolismoRESUMO
Our previous investigation presented evidence of interaction between alpha Ni3S2 and membranous and cellular lipids of lung cells, resulting in significant increases in linoleic, linolenic and arachidonic acids. The present work was designed to follow the metabolic fate of arachidonic acid in alpha Ni3S2-exposed guinea pig alveolar macrophages (GPAM) in culture (50 microM alpha Ni3S2 for 3 days). The metabolites of arachidonic acid were assessed by HPLC coupled with UV or electrochemical detection. The concentrations of malondialdehyde (MDA), hydroxyeicosatetraenoic acid (HETE), leukotrienes (LT) and reduced glutathione (GSH) were measured. In exposed cells a significant increase of MDA, a breakdown product of lipid peroxidation, was observed. In addition, the enzymatic reduction of 5-hydroperoxyeicosatetraenoic acid (5-HPETE) by the associated oxidation of GSH to GSSG increased 5-HETE in GPAM cells and decreased GSH. 5-Hydroperoxyeicosatetraenoic acid was furthermore converted to epoxides, such as leukotriene A4, and we also quantified in exposed cells a significant increase of its subsequent catabolites LTB4, LTC4 and LTE4. Direct measurements of MDA and other metabolites of arachidonic acid clearly show that exposure of GPAM cells to alpha Ni3S2 enhances lipid peroxidation. This lipid peroxidation is an autocatalytic free-radical process and could be responsible for DNA damage. Furthermore, alpha Ni3S2 intoxication induces the release of proinflammatory products, such as leukotrienes, and the decrease of glutathione.
Assuntos
Inflamação/metabolismo , Macrófagos Alveolares/efeitos dos fármacos , Níquel/toxicidade , Animais , Ácido Araquidônico/metabolismo , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Ácidos Graxos Insaturados/metabolismo , Glutationa/metabolismo , Cobaias , Ácidos Hidroxieicosatetraenoicos/metabolismo , Leucotrienos/metabolismo , Malondialdeído/metabolismoRESUMO
Chlorophenols, mainly used as biocides, are compounds with a wide spectrum of toxic effects, including teratogenic and carcinogenic actions. The aim of this study was to examine possible 4-monochlorophenol (4-MCP) toxicity related to metabolic pathways, which may implicate semiquinones and reactive oxygen species (ROS), in human Hep G2 cells. The effects of 4-MCP were performed through cytotoxicity assays (viability, ATP level), metabolic activities (4-MCP intracellular concentration, NADPH cytochrome P-450 reductase (Cyt P-450 red.) and glutathione-S-transferase activities, CYP 3A7 mRNA expression) and oxidative stress (superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase activities, glutathione status, malondialdehyde concentration, CYP 2E1 mRNA expression). According to the literature, in this work Hep G2 cells were incubated in the continuous presence of 4-MCP at 350 microM over 24 or 48 h. Results showed statistically significant decreases in ATP levels (24 or 48 h, P < 0.05) versus controls. The 4-MCP intracellular concentrations increased as early as 8-24 h and then decreased (P < 0.01). Decreases in Cyt. P-450 red. (24 h, P < 0.05), catalase (24 h, P < 0.05; 48 h, P < 0.01), glutathione peroxidase activities (48 h, P < 0.05) and reduced glutathione concentrations (48 h, P < 0.05) were observed. In addition, exposure to 4-MCP increased mRNA expressions of CYP 3A7 (24 h, P < 0.05; 48 h, P < 0.01) and CYP 2E1 (24 h, P < 0.01) versus controls. Taken together, these results suggest that 4-MCP metabolites could induce oxidative stress conditions in Hep G2 cells.
Assuntos
Clorofenóis/toxicidade , Expressão Gênica/efeitos dos fármacos , Neoplasias Hepáticas Experimentais/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Actinas/genética , Animais , Antioxidantes/metabolismo , Hidrocarboneto de Aril Hidroxilases/genética , Carcinoma Hepatocelular , Sobrevivência Celular/efeitos dos fármacos , Citocromo P-450 CYP2E1/genética , Citocromo P-450 CYP3A , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Glutationa/metabolismo , Humanos , Técnicas In Vitro , Neoplasias Hepáticas Experimentais/enzimologia , Malondialdeído/metabolismo , Proteínas/metabolismo , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Testes de Toxicidade , Células Tumorais CultivadasRESUMO
The glycolamide 2-(glycolylamino)fluorene was found to be metabolized in part by induced rat liver microsomes to the hydroxamic acid N-hydroxy-2-(glycolylamino)fluorene. This is the first report of the ability of a microsomal system to carry out the N-hydroxylation of a glycolamide. A comparison of the relative rates of metabolism of the acetyl and glycolyl amides of 2-aminofluorene showed that the former gave about twice as much of the hydroxamic acid as did the latter. On the other hand, the overall metabolism of the glycolamide was slightly more rapid than that of the acetyl congener. Both the glycolyl- and acetyl-derived hydroxamic acids were further metabolized to unknown products by microsomal preparations in the presence of NADPH.
Assuntos
Ácidos Hidroxâmicos/metabolismo , Microssomos Hepáticos/metabolismo , Xenobióticos/metabolismo , Amidas/metabolismo , Animais , Biotransformação , Fluorenos/metabolismo , Hidroxilação , Masculino , Ratos , Ratos EndogâmicosRESUMO
1. Serial bronchoalveolar lavages (BAL) were performed on a subhuman primate (Macacus cynomolgus) in order to give an experimental model for silicosis. 2. We have identified and quantified some plasma proteins of monkey BAL fluid. 3. The results were compared to those previously obtained in humans. 4. Two proteins previously detected in human BAL fluid (alpha 1 acid glycoprotein, alpha 1 antichymotrypsin) were not detected in monkey control BAL fluid. 5. Two kinds of transferrins were detected in monkey BAL fluids while only one is described in human. 6. The present results will now permit sequential follow up studies during the course of experimental silicosis.
Assuntos
Proteínas Sanguíneas/análise , Líquido da Lavagem Broncoalveolar/análise , Macaca fascicularis/metabolismo , Macaca/metabolismo , Animais , Feminino , Humanos , Imunoeletroforese , Inibidores de Proteases/análise , Albumina Sérica/análise , Transferrina/análise , Transferrina/isolamento & purificação , alfa 1-AntitripsinaRESUMO
Exposure to silica can induce fibrosis and/or emphysema. Various factors such as proteases, other hydrolases and oxidants may be involved in the destruction of lung parenchyma. On the other hand, antiproteases play an important role in the protection of lung parenchyma against the action of proteases. We have developed an animal model of silicosis in monkey Macacus cynomolgus and followed these factors by bronchoalveolar lavage (BAL). We have studied glycosidases activities, elastase-like activity, immunoreactive alpha 1-protease inhibitor (alpha 1PI), neutrophil elastase inhibitory capacity (NEIC) and myeloperoxidase. Bronchoalveolar cells in serial BAL were also studied. Six monkeys were exposed to quartz aerosols (100 mg.m-3) for 18 wks. They were followed until they developed X-ray changes, which occurred between 21-64 wks after the end of the dust exposure. Cellular "silicotic nodules" were observed in lung biopsies. A control animal underwent serial BAL. Changes were seen in the differential cell count. The release of superoxide anion by bronchoalveolar cells obtained during the experiment was increased. Separation on a gradient of Percoll showed the presence of young macrophages, which exhibited enhanced release of superoxide anion as compared to the totality of bronchoalveolar cells. The biochemical analysis of BAL fluids obtained during and after the period of dust exposure showed an increase in glycosidases, alpha 1PI and NEIC. Some free elastase-like activity was simultaneously detected in BAL fluids from exposed animals but not from the control. This elastase-like activity was very low compared to NEIC. The increase in enzymatic and antiprotease activities occurred at different points in time for each animal, suggesting large differences in individual responses to dust, but occurred before the chest X-ray abnormalities.
Assuntos
Líquido da Lavagem Broncoalveolar/enzimologia , Líquido da Lavagem Broncoalveolar/patologia , Glicosídeo Hidrolases/metabolismo , Peroxidases/metabolismo , Inibidores de Proteases/metabolismo , Silicose/patologia , Acetilglucosaminidase/metabolismo , Animais , Biópsia , Feminino , Elastase de Leucócito , Pulmão/patologia , Macaca fascicularis , Elastase Pancreática/metabolismo , Silicose/enzimologia , Superóxidos/metabolismo , alfa 1-Antitripsina/metabolismoRESUMO
Available data suggest that repeated concurrent exposure to haematite (Fe(2)O(3)) and benzo[A]pyrene (B[A]P) results in a decreased latency and an increased incidence of lung tumours in rodents compared to exposure to B[A]P alone. Moreover, the reactive oxygen species (ROS) formed by the lung cells themselves and/or by activated inflammatory cells may possibly contribute to the development of pulmonary disorders such as cancer formation. In order to investigate the precise role of iron in the injury induced by B[A]P-coated onto Fe(2)O(3) particles, we tend to address the hypothesis that Fe(2)O(3) and B[A]P, alone or in association, can induce oxidative stress conditions (malondialdehyde) and/or inflammatory reactions (interleukin-6) and thereby disrupt the proteinase/anti-proteinase balance (cathepsins B and L, polynuclear neutrophil (PNN) elastase, alpha-1 proteinase inhibitor (alpha(1)PI) and its inhibitory capacity) in the rat respiratory tract. Thus, Fe(2)O(3) or B[A]P-coated onto Fe(2)O(3) particles produce oxidative stress conditions through not only iron-catalysed oxidative reactions but also inflammatory processes. However, B[A]P initiates only inflammatory responses. These pollutants generate increased levels of proteases and decrease the concentrations of free alpha(1)PI. There is also a clear relationship between the partial inactivation of alpha(1)PI and the occurrence of ROS after exposure to Fe(2)O(3), alone or as a carrier of B[A]P. Hence, the proteinase/anti-proteinase balance might be more disrupted by Fe(2)O(3) or B[A]P-coated onto Fe(2)O(3) particles than by B[A]P alone. These results suggest a mechanism that can explain why B[A]P-coated onto Fe(2)O(3) particles are more injurious than B[A]P alone.