RESUMO
CONTEXT: Sulfur mustard is a blister agent that can cause death by pulmonary damage. There is currently no effective treatment. N-acetyl-L-cysteine (NAC) has mucolytic and antioxidant actions and is an important pre-cursor of cellular glutathione synthesis. These actions may have potential to reduce mustard-induced lung injury. OBJECTIVE: Evaluate the effect of nebulised NAC as a post-exposure treatment for inhaled sulfur mustard in a large animal model. MATERIALS AND METHODS: Fourteen anesthetized, surgically prepared pigs were exposed to sulfur mustard vapor (100 µg.kg⻹), 10 min) and monitored, spontaneously breathing, to 12 h. Control animals had no further intervention (n = 6). Animals in the treatment group were administered multiple inhaled doses of NAC (1 ml of 200 mg.ml⻹ Mucomyst™ at + 30 min, 2, 4, 6, 8, and 10 h post-exposure, n = 8). Cardiovascular and respiratory parameters were recorded. Arterial blood was collected for blood gas analysis while blood and bronchoalveolar lavage fluid were collected for hematology and inflammatory cell analysis. Urine was collected to detect a sulfur mustard breakdown product. Lung tissue samples were taken for histopathological and post-experimental analyses. RESULTS: Five of six sulfur mustard-exposed animals survived to 12 h. Arterial blood oxygenation (PaO2) and saturation levels were significantly decreased at 12 h. Arterial blood carbon dioxide (PaCO2) significantly increased, and arterial blood pH and bicarbonate (HCO3â») significantly decreased at 12 h. Shunt fraction was significantly increased at 12 h. In the NAC-treated group all animals survived to 12 h (n = 8). There was significantly improved arterial blood oxygen saturation, HCO3â» levels, and shunt fraction compared to those of the sulfur mustard controls. There were significantly fewer neutrophils and lower concentrations of protein in lavage compared to sulfur mustard controls. DISCUSSION: NAC's mucolytic and antioxidant properties may be responsible for the beneficial effects seen, improving clinically relevant physiological indices affected by sulfur mustard exposure. CONCLUSION: Beneficial effects of nebulized NAC were apparent following inhaled sulfur mustard exposure. Further therapeutic benefit may result from a combination therapy approach.
Assuntos
Acetilcisteína/uso terapêutico , Antioxidantes/uso terapêutico , Substâncias para a Guerra Química/toxicidade , Modelos Animais de Doenças , Intoxicação por Gás/tratamento farmacológico , Pulmão/efeitos dos fármacos , Gás de Mostarda/toxicidade , Acetilcisteína/administração & dosagem , Acetilcisteína/efeitos adversos , Administração por Inalação , Aerossóis , Animais , Antídotos/administração & dosagem , Antídotos/efeitos adversos , Antídotos/uso terapêutico , Antioxidantes/administração & dosagem , Antioxidantes/efeitos adversos , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/imunologia , Substâncias para a Guerra Química/análise , Substâncias para a Guerra Química/farmacocinética , Expectorantes/administração & dosagem , Expectorantes/efeitos adversos , Expectorantes/uso terapêutico , Feminino , Intoxicação por Gás/imunologia , Intoxicação por Gás/patologia , Intoxicação por Gás/fisiopatologia , Pulmão/imunologia , Pulmão/patologia , Pulmão/fisiopatologia , Pneumopatias/etiologia , Pneumopatias/prevenção & controle , Gás de Mostarda/administração & dosagem , Gás de Mostarda/análise , Gás de Mostarda/farmacocinética , Infiltração de Neutrófilos/efeitos dos fármacos , Distribuição Aleatória , Insuficiência Respiratória/etiologia , Insuficiência Respiratória/prevenção & controle , Análise de Sobrevida , Sus scrofaRESUMO
CONTEXT: Inhalation of sulfur mustard (HD) vapor can cause life-threatening lung injury for which there is no specific treatment. A reproducible, characterized in vivo model is required to investigate novel therapies targeting HD-induced lung injury. MATERIALS AND METHODS: Anesthetized, spontaneously breathing large white pigs (~50 kg) were exposed directly to the lung to HD vapor at 60, 100, or 150 µg/kg, or to air, for ~10 min, and monitored for 6 h. Cardiovascular and respiratory parameters were recorded. Blood and bronchoalveolar lavage fluid (BALF) were collected to allow blood gas analysis, hematology, and to assay for lung inflammatory cells and mediators. Urine was collected and analyzed for HD metabolites. Histopathology samples were taken postmortem (PM). RESULTS: Air-exposed animals maintained normal lung physiology whilst lying supine and spontaneously breathing. There was a statistically significant increase in shunt fraction across all three HD-exposed groups when compared with air controls at 3-6 h post-exposure. Animals were increasingly hypoxemic with respiratory acidosis. The monosulfoxide ß-lyase metabolite of HD (1-methylsulfinyl-2-[2(methylthio)ethylsulfonyl)ethane], MSMTESE), was detected in urine from 2 h post-exposure. Pathological examination revealed necrosis and erosion of the tracheal epithelium in medium and high HD-exposed groups. CONCLUSION: These findings are consistent with those seen in the early stages of acute lung injury (ALI).
Assuntos
Modelos Animais de Doenças , Exposição por Inalação/efeitos adversos , Gás de Mostarda/administração & dosagem , Gás de Mostarda/toxicidade , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/patologia , Fatores Etários , Animais , Relação Dose-Resposta a Droga , Feminino , Gás de Mostarda/metabolismo , Oxiemoglobinas/metabolismo , Mucosa Respiratória/efeitos dos fármacos , Mucosa Respiratória/patologia , Suínos , Fatores de TempoRESUMO
ABSTRACT Although normally regarded as a vesicant, inhalation of sulphur mustard (HD) vapor can cause life-threatening lung injury for which there is no specific treatment. Novel therapies for HD-induced lung injury are best investigated in an in vivo model that allows monitoring of a range of physiological variables. HD vapor was generated using two customized thermostatically controlled glass flasks in parallel. The vapor was passed into a carrier flow of air (81 L. min(-1)) and down a length of glass exposure tube (1.75 m). A pig was connected to the midpoint of the exposure tube via a polytetrafluoroethylene-lined endotracheal tube, Fleisch pneumotachograph, and sample port. HD vapor concentrations (40-122.8 mg. m(-3)) up-and downstream of the point of exposure were obtained by sampling onto Porapak absorption tubes with subsequent analysis by gas chromatography-flame photometric detection. Real-time estimates of vapor concentration were determined using a photo-ionization detector. Lung function indices (respiratory volumes, lung compliance, and airway resistance) were measured online throughout. Trial runs with methylsalicylate (MS) and animal exposures with HD demonstrated that the exposure system rapidly reached the desired concentration within 1 min and maintained stable output throughout exposure, and that the MS/HD concentration decayed rapidly to zero when switched off. A system is described that allows reproducible exposure of HD vapor to the lung of anesthetized white pigs. The system has proved to be robust and reliable and will be a valuable tool in assessing potential future therapies against HD-induced lung injury in the pig. Crown Copyright (c) 2007 Dstl.
RESUMO
A workshop was held February 14, 2007, in Arlington, VA, under the auspices of the Phosgene Panel of the American Chemistry Council. The objective of this workshop was to convene inhalation toxicologists and medical experts from academia, industry and regulatory authorities to critically discuss past and recent inhalation studies of phosgene in controlled animal models. This included presentations addressing the benefits and limitations of rodent (mice, rats) and nonrodent (dogs) species to study concentration x time (C x t) relationships of acute and chronic types of pulmonary changes. Toxicological endpoints focused on the primary pulmonary effects associated with the acute inhalation exposure to phosgene gas and responses secondary to injury. A consensus was reached that the phosgene-induced increased pulmonary extravasation of fluid and protein can suitably be probed by bronchoalveolar lavage (BAL) techniques. BAL fluid analyses rank among the most sensitive methods to detect phosgene-induced noncardiogenic, pulmonary high-permeability edema following acute inhalation exposure. Maximum protein concentrations in BAL fluid occurred within 1 day after exposure, typically followed by a latency period up to about 15 h, which is reciprocal to the C x t exposure relationship. The C x t relationship was constant over a wide range of concentrations and single exposure durations. Following intermittent, repeated exposures of fixed duration, increased tolerance to recurrent exposures occurred. For such exposure regimens, chronic effects appear to be clearly dependent on the concentration rather than the cumulative concentration x time relationship. The threshold C x t product based on an increased BAL fluid protein following single exposure was essentially identical to the respective C x t product following subchronic exposure of rats based on increased pulmonary collagen and influx of inflammatory cells. Thus, the chronic outcome appears to be contingent upon the acute pulmonary threshold dose. Exposure concentrations high enough to elicit an increased acute extravasation of plasma constituents into the alveolus may also be associated with surfactant dysfunction, intra-alveolar accumulation of fibrin and collagen, and increased recruitment and activation of inflammatory cells. Although the exact mechanisms of toxicity have not yet been completely elucidated, consensus was reached that the acute pulmonary toxicity of phosgene gas is consistent with a simple, irritant mode of action at the site of its initial deposition/retention. The acute concentration x time mortality relationship of phosgene gas in rats is extremely steep, which is typical for a local, directly acting pulmonary irritant gas. Due to the high lipophilicity of phosgene gas, it efficiently penetrates the lower respiratory tract. Indeed, more recent published evidence from animals or humans has not revealed appreciable irritant responses in central and upper airways, unless exposure was to almost lethal concentrations. The comparison of acute inhalation studies in rats and dogs with focus on changes in BAL fluid constituents demonstrates that dogs are approximately three to four times less susceptible to phosgene than rats under methodologically similar conditions. There are data to suggest that the dog may be useful particularly for the study of mechanisms associated with the acute extravasation of plasma constituents because of its size and general morphology and physiology of the lung as well as its oronasal breathing patterns. However, the study of the long-term sequelae of acute effects is experimentally markedly more demanding in dogs as compared to rats, precluding the dog model to be applied on a routine base. The striking similarity of threshold concentrations from single exposure (increased protein in BAL fluid) and repeated-exposure 3-mo inhalation studies (increased pulmonary collagen deposition) in rats supports the notion that chronic changes depend on acute threshold mechanisms.
Assuntos
Modelos Animais de Doenças , Pulmão/efeitos dos fármacos , Fosgênio/toxicidade , Animais , Biomarcadores/metabolismo , Líquido da Lavagem Broncoalveolar/química , Cães , Relação Dose-Resposta a Droga , Humanos , Pulmão/metabolismo , Pulmão/patologia , Pneumopatias/induzido quimicamente , Pneumopatias/metabolismo , Pneumopatias/patologia , Camundongos , Ratos , Especificidade da Espécie , Fatores de Tempo , VirginiaRESUMO
ABSTRACT Guinea pigs are considered as the animal model of choice for toxicology and medical countermeasure studies against chemical warfare agents (CWAs) and toxic organophosphate pesticides because of the low levels of carboxylesterase compared to rats and mice. However, it is difficult to intubate guinea pigs without damaging the larynx to perform CWA inhalation experiments. We describe an easy technique of intubation of guinea pigs for accurate endotracheal placement of the intubation tube. The technique involves a speculum made by cutting the medium-size ear speculum in the midline leaving behind the intact circular connector to the otoscope. Guinea pigs were anesthetized with Telazol/meditomidine, the tongue was pulled using blunt forceps, and an otoscope attached with the specially prepared speculum was inserted gently. Insertion of the speculum raises the epiglottis and restrains the movements of vocal cord, which allows smooth insertion of the metal stylet-reinforced intubation tube. Accurate endotracheal placement of the intubation tube was achieved by measuring the length from the tracheal bifurcation to vocal cord and vocal cord to the upper front teeth. The average length of the trachea in guinea pigs (275 +/- 25 g) was 5.5 +/- 0.2 cm and the distance from the vocal cord to the front teeth was typically 3 cm. Coinciding an intubation tube marked at 6 cm with the upper front teeth accurately places the intubation tube 2.5 cm above the tracheal bifurcation. This simple method of intubation does not disturb the natural flora of the mouth and causes minimum laryngeal damage. It is rapid and reliable, and will be very valuable in inhalation exposure to chemical/biological warfare agents or toxic chemicals to assess respiratory toxicity and develop medical countermeasures.
RESUMO
The use of chemical warfare agents, such as the pulmonary irritant gas phosgene, is a real and constant threat not only from belligerent nations but from terrorist groups as well. Phosgene is both easy and inexpensive to produce and as such is a potential candidate for use as a threat agent. Phosgene attacks the deep lung after inhalation and can severely compromise pulmonary mechanics and gas exchange, rendering the exposed individual incapacitated. If exposure is severe, death can ensure by asphyxiation secondary to pulmonary edema formation. This paper examines the effects on lung tissue in mice over 24 hours after exposure to the irritant gas phosgene. Exposure to phosgene produced respiratory acidosis by decreasing pH, partial pressure of oxygen, O2 saturation, and increasing partial pressure of carbon dioxide. Exposure to phosgene also induced temporal increases in lung tissue gravimetric parameters such as lung tissue wet weight/dry weight ratio, which is a positive indicator of pulmonary edema formation, and dry lung weight, an indicator of lung cellular hyperaggregation. Blood gases and pH tend to normalize within 24 hours, whereas gravimetric parameters remain increased. Temporal changes in these physiological indicators of lung injury may help to explain why past exposures to phosgene required lengthy hospitalization.
Assuntos
Substâncias para a Guerra Química/efeitos adversos , Pulmão/efeitos dos fármacos , Fosgênio/efeitos adversos , Troca Gasosa Pulmonar/efeitos dos fármacos , Análise de Variância , Animais , Pulmão/fisiopatologia , Masculino , CamundongosRESUMO
Phosgene, widely used in industrial processes, can cause life-threatening pulmonary edema and acute lung injury. One mechanism of protection against phosgene-induced lung injury may involve the use of antioxidants. The present study focused on dietary supplementation in mice using n-propyl gallate (nPG)--a gallate acid ester compound used in food preservation--and vitamin E. Five groups of male mice were studied: group 1, control-fed with Purina rodent chow 5002; group 2, fed 0.75% nPG (w/w) in 5002; group 3, fed 1.5% nPG (w/w) in 5002; group 4 fed 1% (w/w) vitamin E in 5002; and group 5, fed 2% (w/w) vitamin E also in 5002. Mice were fed for 23 days. On day 23 mice were exposed to 32 mg m-3 (8 ppm) phosgene for 20 min (640 mg. min m-3) in a whole-body exposure chamber. Survival rates were determined at 12 and 24 h. In mice that died within 12 h, the lungs were removed and lung wet weights, dry weights, wet/dry weight ratios, lipid peroxidation (thiobarbituric acid reactive substances, TBARS) and glutathione (GSH) were assessed. Vitamin E had no positive effect on any outcome measured. There was no significant difference between 1.5% nPG and any parameter measured or survival rate compared with 5002 + phosgene. However, dietary treatment with 0.75% nPG significantly increased survival rate (P = 0.002) and lowered TBARS (P = 0.05) compared with 5002 + phosgene at 12 h after exposure. Mice fed 0.75% nPG had a lower wet/dry wt ratio compared with those fed 1.5% nPG and a significantly increased lung tissue GSH 36%, compared with the 5002 + phosgene group. In conclusion, dietary treatment with a low level of the antioxidant nPG protected mice by decreasing lipid peroxidation and increasing lung tissue GSH. The higher level of nPG and both levels of vitamin E diets were ineffective, suggesting that a ceiling threshold level of antioxidants in lung tissue is required for survival against phosgene-induced lung injury. Published in 2001 by John Wiley & Sons, Ltd.
Assuntos
Antioxidantes/administração & dosagem , Fosgênio/toxicidade , Galato de Propila/administração & dosagem , Edema Pulmonar/dietoterapia , Vitamina E/administração & dosagem , Administração por Inalação , Animais , Dieta , Glutationa/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Camundongos , Tamanho do Órgão/efeitos dos fármacos , Fosgênio/administração & dosagem , Edema Pulmonar/induzido quimicamente , Edema Pulmonar/mortalidade , Edema Pulmonar/prevenção & controle , Taxa de Sobrevida , Fatores de TempoRESUMO
Exposure to phosgene has been shown to cause severe and life-threatening pulmonary edema. There is evidence that successful treatment of phosgene-induced acute lung injury may be related to increased antioxidant activity. Acetylenic acids such as 5,8,11, 14-eicosatetraynoic acid (ETYA) have been shown to be effective in preventing pulmonary edema formation (PEF). In phosgene-exposed guinea pigs, we examined the effects of ETYA on PEF. Lipid peroxidation (thiobarbituric acid-reactive substance, TBARS) and total glutathione (GSH) were measured in lung tissue from isolated, buffer-perfused guinea pig lungs at 180 min after start of exposure. Guinea pigs were challenged with 175 mg/m(3) (44 ppm) phosgene for 10 min (1750 mg( small middle dot)min/m(3)). Five minutes after removal from the exposure chamber, guinea pigs were treated, ip, with 200 microl of 100 microM ETYA in ethanol (ETOH). Two hundred microliters of 50 microM ETYA in ETOH was added to the 200 ml perfusate every 40 min beginning at 60 min after start of exposure (t = 0). There were four groups in this study: air-exposed, phosgene-exposed, phosgene + ETYA-posttreated, and air + ETYA-posttreated. Posttreatment with ETYA prevented GSH depletion, 2. 7 +/- 0.5 micromol/mg protein versus 1 +/- 0.2 micromol/mg protein, for the untreated phosgene-exposed lungs (p < or =.05). ETYA posttreatment also significantly decreased PEF (p =.025), as measured by lung wet weight/dry weight ratio, 16.1 +/- 2.5 versus 8.5 +/- 1 for phosgene-exposed + ETYA-posttreated guinea pigs. Postexposure treatment with ETYA significantly increased the GSH to TBARS protection ratio, 12 +/- 2, compared with the phosgene-exposed group, 3.7 +/- 0.5 (p < or =.05). In conclusion, ETYA posttreatment decreased PEF by increasing the GSH/TBARS protection ratio by functioning in an antioxidant-like capacity.
Assuntos
Ácido 5,8,11,14-Eicosatetrainoico/uso terapêutico , Glutationa , Peroxidação de Lipídeos/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Fosgênio/toxicidade , Edema Pulmonar/prevenção & controle , Ácido 5,8,11,14-Eicosatetrainoico/administração & dosagem , Administração por Inalação , Animais , Glutationa/metabolismo , Cobaias , Injeções Intraperitoneais , Pulmão/metabolismo , Pulmão/patologia , Masculino , Tamanho do Órgão/efeitos dos fármacos , Fosgênio/administração & dosagem , Edema Pulmonar/induzido quimicamenteRESUMO
Phosgene-induced pulmonary edema formation has been under investigation for many years. One mechanism of protection may involve the use of antioxidants. Previously, it has been shown that butylated hydroxyanisole (BHA) treatment can enhance glutathione (GSH) levels. The present study focused on dietary supplementation in mice using BHA, a phenolic compound used in food preservation. Three groups of male CD-1 mice were studied: group 1, control animals fed with Purina rodent chow 5002; group 2, fed 0.75% BHA (w/w) in 5002; and group 3, fed 1.5% BHA (w/w) in 5002. Mice were fed for 22 days. On day 23 mice were exposed to 32 mg/m(3) phosgene for 20 min in a whole-body exposure chamber. Survival rate (SR) and odds ratio (OR) were determined at 12 and 24 h. In mice that died within 12 h, the lungs were removed immediately and lung wet weights (WW), dry weights (DW), lung wet weight/body weight ratio (LWW/BW), and lung tissue total glutathione (GSH) were assessed. For 12-h data, 6 mice from the 1.5% BHA group were sacrificed for lung tissue measurements. The SR for 0.75% BHA was 80% at 12 h and 55% at 24 h, compared with 36% and 23%, respectively, for controls. For 1.5% BHA, the 12- and 24-h SR were 100% and 92%, respectively. Odds ratios of 6.9 for 0.75% BHA and 46.6 for 1.5% BHA at 12 h and 4.0 and 42 for 0. 75% and 1.5% BHA, respectively, at 24 h were significantly (chi2) higher than control diet phosgene-exposed mice. Dietary pretreatment with 0.75% and 1.5% BHA significantly enhanced lung tissue GSH, 1.8-fold (p < or =.01) and 5.8-fold (p < or =.01), respectively, compared with phosgene-exposed control diet. Both BHA-supplemented diets significantly reduced WW. Only 1.5% BHA reduced DW, a measure of lung hyperaggregation. and LWW/BW compared with control diet. In air-exposed controls, BHA induced a dose-responsive decrease in WW, DW, LWW/BW ratio, and GSH. In conclusion, dietary pretreatment with BHA at the two dose levels reduced lung edema and lethality by enhancing lung tissue GSH in mice exposed to phosgene.
Assuntos
Antioxidantes/farmacologia , Hidroxianisol Butilado/farmacologia , Substâncias para a Guerra Química/toxicidade , Glutationa/metabolismo , Pulmão/metabolismo , Fosgênio/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Dieta , Pulmão/efeitos dos fármacos , Masculino , Camundongos , Tamanho do Órgão/efeitos dos fármacosRESUMO
This study investigated the post-treatment effect of isoproterenol (ISO) on pulmonary parameters in rabbits whole-body-exposed to a lethal dose of the toxic gas phosgene. Phosgene is widely used in industry as a chemical intermediate for the production of plastics, drugs and polyurethane products. The results of this study are from five study groups: 10-min perfused baseline; uninjured controls exposed to air; phosgene-exposed; phosgene-exposed isoproterenol-treated intravascularly and intratracheally (ISO i.v.+i.t.); and phosgene-exposed isoproterenol-treated intratracheally (ISO i.t.). Treatment with ISO was administered as either a continuous intravascular infusion (24 microg min(-1)) from the beginning to end of perfusion (i.v.) and a 24-microg intratracheal bolus (i.t.) or just an i.t. bolus immediately prior to the start of perfusion. Rabbits of 2.5-3 kg were exposed to a cumulative dose of phosgene to attain a concentration x time exposure-effect of 1500 ppm x min. Lungs were isolated in situ and perfused 50-60 min after the start of exposure with Krebs-Henseleit buffer at 40 ml min(-1). Pulmonary artery pressure (Ppa), tracheal pressure (Pt) and lung weight gain (lwg) were continuously measured. Leukotrienes (LT) C4/D4/E4 were measured in the perfusate every 20 min during perfusion. At the immediate conclusion of the experiment, lung tissue was frozen in liquid N2 and analyzed for glutathione (GSH) and cyclic 3',5'-adenosine monophosphate (cAMP). Post-treatment with ISO by either i.v.+i.t. or i.t. routes 50+ min after phosgene exposure significantly lowered Ppa, Pt and lwg. Phosgene-exposed rabbits post-treated with ISO i.t. had significantly higher levels of reduced GSH (3 +/- 0.4 nmol mg(-1) protein), GSH/GSSG ratios (3.3 +/- 0.6 nmol mg(-1) protein) and percentage of total as reduced GSH (75 +/- 2.5%) compared with phosgene-exposed rabbits: 1.9 +/- 0.3, 2 +/- 0.3 and 58 +/- 6.3%, respectively. The ISO (i.v.+i.t.) post-treatment route significantly increased reduced GSH (6.2 +/- 1.7 nmol mg(-1) protein), GSH/GSSG ratio (5.9 +/- 0.8 nmol mg(-1) protein) and percentage of total as reduced GSH (85 +/- 1.7%) when compared to the phosgene-only group. The ISO i.t. and ISO i.v.+i.t. treatments significantly reduced perfusate LTC4/D4/E4 150 min after the start of exposure by 90% and 48%, respectively. These data suggest that protective mechanisms for ISO involve reduced vascular pressure, decreased LTC4/D4/E4-mediated pulmonary capillary permeability and a favorable lung tissue redox state compared with untreated phosgene-exposed rabbits.
Assuntos
Isoproterenol/uso terapêutico , Fosgênio/intoxicação , Edema Pulmonar/tratamento farmacológico , Animais , AMP Cíclico/análise , Glutationa/análise , Isoproterenol/administração & dosagem , Leucotrieno C4/análise , Leucotrieno D4/análise , Leucotrieno E4/análise , Pulmão/metabolismo , Masculino , Edema Pulmonar/induzido quimicamente , Edema Pulmonar/metabolismo , Coelhos , Fatores de TempoRESUMO
Acetylenic acids such as 5,8,11,14-eicosatetraynoic acid (ETYA), have been shown to be effective in preventing pulmonary edema formation (PEF). In phosgene-exposed guinea pigs, we examined the effects of ETYA on PEF, measured as real time lung weight gain (lwg). Pulmonary artery pressure (Ppa), airway pressure (Paw), perfusate leukotrienes (LT) C4/D4/E4/B4, and lung tissue lipid peroxidation (TBARS) were measured using the isolated, buffer-perfused lung model. Guinea pigs were challenged to 175 mg/m3 (44 ppm) phosgene for 10 minutes giving a concentration x time product of 1750 mg.min/m3 (437 ppm.min). Five minutes after removal from the exposure chamber, guinea pigs were treated, i.p., with 200 microL of 100 microM ETYA. 200 microL of 50 microM ETYA was added to the perfusate every 40 minutes, beginning at 60 minutes after start of exposure (t = 0). There were four groups in this study: air-treated, phosgene-exposed, ETYA-posttreated + phosgene, and ETYA-posttreated + air ETYA-posttreated + phosgene guinea pigs had significantly lower Ppa (P = .006), Paw (P = .009), and lwg (P = .016) compared with phosgene-exposed animals. Phosgene exposure reduced LTB4 compared with air-treated controls (P = .09). ETYA-posttreatment + phosgene had significantly increased perfusate LTB4 (P = .0006) compared with phosgene exposure only group. Total perfusate, LTC4 + LTD4 + LTE4, was not different between phosgene-exposed, air-treated or ETYA-posttreatment + phosgene over time. Posttreatment with ETYA significantly lowered TBARS formation, 206 +/- 13 versus 285 +/- 23 nmol/mg protein (P = .016), compared with phosgene-exposed lungs. Paradoxically, ETYA posttreatment decreased PEF and lipid peroxidation, but increased sulfidopeptide LT release from the lung during perfusion. We conclude that LTC4/D4/E4, and B4, may play different roles than previously thought for PEF in the isolated perfused lung model.
Assuntos
Ácido 5,8,11,14-Eicosatetrainoico/farmacologia , Leucotrienos/metabolismo , Pulmão/efeitos dos fármacos , Fosgênio/toxicidade , Edema Pulmonar/prevenção & controle , Resistência das Vias Respiratórias/efeitos dos fármacos , Animais , Pressão Sanguínea/efeitos dos fármacos , Cobaias , Técnicas In Vitro , Peroxidação de Lipídeos/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Tamanho do Órgão , Perfusão , Artéria Pulmonar/efeitos dos fármacos , Artéria Pulmonar/fisiologia , Edema Pulmonar/induzido quimicamente , Edema Pulmonar/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
Phosgene is a highly reactive oxidant gas used in the chemical industry. Phosgene can cause life-threatening pulmonary edema by reacting with peripheral lung compartment tissue components. Clinical evidence of edema is not usually apparent until well after the initial exposure. This study was designed to investigate early signs of acute lung injury in rodents within 45-60 min after the start of exposure. Male mice, rats, or guinea pigs were exposed to 87 mg/m3 (22 ppm) phosgene or filtered room air for 20 min followed by room air washout for 5 min. This concentration-time exposure causes a doubling of lung wet weight within 5 h. After exposure, animals were immediately anesthetized i.p., with pentobarbital. Bronchoalveolar lavage (BAL) was performed and fluid analyzed for total glutathione (GSH), lipid peroxidation thiobarbituric acid reactive substances (TBARS), and protein concentration. Lungs were perfused with saline to remove blood, freeze-snapped in liquid N2, analyzed for tissue GSH, and TBARS. Lung edema was assessed gravimetrically by measuring tissue wet/dry (W/D) weight ratios and tissue wet weights (TWW). W/D and TWW were significantly higher in mice for phosgene vs air (P=0.001, P < 0.0001, respectively), but not in rats or guinea pigs. Tissue TBARS was significantly higher in phosgene-exposed guinea pigs, P=0.027; however, BAL TBARS was higher in both rats and guinea pigs, P=0.013 and P=0.006, respectively. Tissue GSH was significantly lower in phosgene-exposed rats and guinea pigs but not mice, whereas BAL GSH was higher in rats, P < 0.0001. There were significantly higher BAL protein levels in all phosgene-exposed species: mice, P < 0.0001; rats, P < 0.0001; and guinea pigs, P=0.002. Although there appears to be a species-specific biochemical effect of phosgene exposure for some biochemical indices, measurement of BAL protein in all three species is a better indicator of ensuing edema formation.
Assuntos
Líquido da Lavagem Broncoalveolar/química , Peroxidação de Lipídeos/efeitos dos fármacos , Fosgênio/toxicidade , Proteínas/análise , Edema Pulmonar/induzido quimicamente , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Doença Aguda , Animais , Estudos de Avaliação como Assunto , Glutationa/análise , Cobaias , Masculino , Camundongos , Camundongos Endogâmicos ICR , Ratos , Ratos Sprague-DawleyRESUMO
Pretreatment with aminophylline has been shown to protect against various types of acute lung injury. Mechanisms responsible for protection are multifactorial but are thought to involve upregulation of cAMP. While previous studies focused on pretreatment, the present investigation examined post-treatment in rabbits following exposure to a lethal dose of the oxidant gas phosgene. Rabbits, 2-3 kg, were exposed to a cumulative dose of phosgene to attain a c x t exposure effect of 1500 ppm.min. Lungs were isolated in situ and perfused for 90-100 min after exposure with Krebs-Henseleit buffer at 40 mL/min. Pulmonary artery pressure (Ppa), tracheal pressure (Pt), and lung weight gain (lwg) were measured continuously. Leukotrienes C4/D4/E4 were measured in the perfusate every 20 min during perfusion. At the immediate conclusion of the experiment, lung tissue was frozen in liquid N2 and analyzed for reduced GSH, GSSG, cAMP, and lipid peroxidation (TBARS). Post-treatment with aminophylline 80-90 min after exposure significantly lowered Ppa, Pt, and lwg. Aminophylline significantly reduced TBARS and perfusate LTC4/D4/E4, and prevented phosgene-induced decreases in lung tissue cAMP. These data suggest that protective mechanisms observed with aminophylline involve decreased LTC4/D4/E4-mediated pulmonary capillary permeability and attenuated lipid peroxidation. Direct antipermeability effects of cAMP on cellular contraction may also be important in protection against phosgene-induced lung injury.
Assuntos
Aminofilina/farmacologia , Substâncias para a Guerra Química/toxicidade , Pneumopatias/prevenção & controle , Fosgênio/toxicidade , Inibidores de Fosfodiesterase/farmacologia , Administração por Inalação , Animais , Pressão Sanguínea/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Pulmão/fisiopatologia , Pneumopatias/induzido quimicamente , Pneumopatias/fisiopatologia , Masculino , Tamanho do Órgão/efeitos dos fármacos , Perfusão , Artéria Pulmonar/efeitos dos fármacos , Artéria Pulmonar/fisiologia , Coelhos , Testes de Função Respiratória , SRS-A/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
Mice, rats and guinea pigs were exposed to phosgene (COCl2), a highly irritating and oxidizing gas. Animals were exposed to 87 mg/m3 phosgene for 20 min in a whole-body exposure chamber. Within 55-65 minutes after the start of exposure, plasma was scanned spectrophotometrically from 200-600 nm. A distinct and significant increase in area under the curve in the Soret band region at 413 nm was observed in plasma from phosgene-exposed animals when compared with air-exposed controls in all three species. These peaks were consistent with hemoglobin, an indication that the integrity of the erythrocyte membrane had been compromised by exposure. An erythrocyte osmotic fragility assay on blood from mice exposed to phosgene indicated that 30% less NaCl was needed to cause 50% hemolysis compared to air-exposed mice. These results suggest a new mechanism of phosgene-induced acute lung injury that may be linked, in part, to a direct attack on the erythrocyte membrane.
Assuntos
Sangue/efeitos dos fármacos , Fosgênio/toxicidade , Animais , Membrana Eritrocítica/efeitos dos fármacos , Membrana Eritrocítica/fisiologia , Cobaias , Hemólise , Masculino , Camundongos , Camundongos Endogâmicos , Fragilidade Osmótica/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Valores de Referência , EspectrofotometriaRESUMO
Phosgene, a highly reactive former warfare gas, is a deep lung irritant which produces adult respiratory distress syndrome (ARDS)-like symptoms following inhalation. Death caused by phosgene involves a latent, 6-24-h, fulminating non-cardiogenic pulmonary edema. The following dose-ranging study was designed to determine the efficacy of a non-steroidal anti-inflammatory drug, ibuprofen (IBU), and a methylxanthine, pentoxifylline (PTX). These drugs were tested singly and in combination to treat phosgene-induced acute lung injury in rats. Ibuprofen, in concentrations of 15-300 mg kg-1 (i.p.), was administered to rats 30 min before and 1 h after the start of whole-body exposure to phosgene (80 mg m-3 for 20 min). Pentoxifylline, 10-120 mg kg-1 (i.p.), was first administered 15 min prior to phosgene exposure and twice more at 45 and 105 min after the start of exposure. Five hours after phosgene inhalation, rats were euthanized, the lungs were removed and wet weight values were determined gravimetrically. Ibuprofen administered alone significantly decreased lung wet weight to body weight ratios compared with controls (P < or = 0.01) whereas PTX, at all doses tested alone, did not. In addition, the decrease in lung wet weight to body weight ratio observed with IBU+PTX could be attributed entirely to the dose of IBU employed. This is the first study to show that pre- and post-treatment with IBU can significantly reduce lung edema in rats exposed to phosgene.
Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Ibuprofeno/farmacologia , Pulmão/efeitos dos fármacos , Pentoxifilina/farmacologia , Inibidores de Fosfodiesterase/farmacologia , Administração por Inalação , Análise de Variância , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/uso terapêutico , Substâncias para a Guerra Química/toxicidade , Modelos Animais de Doenças , Sinergismo Farmacológico , Ibuprofeno/administração & dosagem , Ibuprofeno/uso terapêutico , Injeções Intraperitoneais , Lesão Pulmonar , Masculino , Tamanho do Órgão/efeitos dos fármacos , Pentoxifilina/administração & dosagem , Pentoxifilina/uso terapêutico , Fosgênio/administração & dosagem , Fosgênio/toxicidade , Inibidores de Fosfodiesterase/administração & dosagem , Inibidores de Fosfodiesterase/uso terapêutico , Edema Pulmonar/induzido quimicamente , Edema Pulmonar/tratamento farmacológico , Edema Pulmonar/mortalidade , Ratos , Ratos Sprague-Dawley , Síndrome do Desconforto Respiratório/induzido quimicamenteRESUMO
Phosgene, a toxic gas widely used as an industrial chemical intermediate, is known to cause life-threatening latent noncardiogenic pulmonary edema. Mechanisms related to its toxicity appear to involve lipoxygenase mediators of arachidonic acid (AA) and can be inhibited by pretreatment with drugs that increase adenosine 3',5'-cyclic monophosphate (cAMP). In the present study, we used the isolated buffer-perfused rabbit lung model to investigate the mechanisms by which cAMP protects against phosgene-induced lung injury. Posttreatment with dibutyryl cAMP (DBcAMP) was given 60-85 min after exposure by an intravascular or intratracheal route. Lung weight gain (LWG) was measured continuously. AA metabolites leukotriene (LT) C4, LTD4, and LTE4 and 6-ketoprostaglandin F1 alpha were measured in the perfusate at 70, 90, 110, 130, and 150 min after exposure. Tissue malondialdehyde and reduced and oxidized glutathione were analyzed 150 min postexposure. Compared with measurements in the lungs of rabbits exposed to phosgene alone, posttreatment with DBcAMP significantly reduced LWG, pulmonary arterial pressure, and inhibited the release of LTC4, LTD4, and LTE4. Intratracheal administration of DBcAMP was more effective than intravascular administration in reducing LWG. Posttreatment also decreased MDA and protected against glutathione oxidation observed with phosgene exposure. We conclude that phosgene causes marked glutathione oxidation, lipid peroxidation, release of AA mediators, and increases LWG. Posttreatment with DBcAMP attenuates these effects, not only by previously described inhibition of pulmonary endothelial or epithelial cell contraction but also by inhibition of AA-mediator production and a novel antioxidant effect.
Assuntos
Bucladesina/farmacologia , Fosgênio , Edema Pulmonar/tratamento farmacológico , 6-Cetoprostaglandina F1 alfa/metabolismo , Animais , Ácidos Araquidônicos/metabolismo , Bucladesina/administração & dosagem , Glutationa , Técnicas In Vitro , Injeções Intravenosas , Intubação Intratraqueal , Leucotrienos/metabolismo , Masculino , Malondialdeído/metabolismo , Tamanho do Órgão/efeitos dos fármacos , Tamanho do Órgão/fisiologia , Proteínas/metabolismo , Edema Pulmonar/induzido quimicamente , Edema Pulmonar/metabolismo , CoelhosRESUMO
We examined the effects of treatment with N-acetylcysteine (NAC) on pulmonary edema formation in isolated perfused rabbit lungs following in vivo phosgene exposure. This study focused on posttreatment intratracheal administration of NAC after exposure. Rabbits, 2 to 3 kg, were exposed to a cumulative dose of phosgene to attain a concentration x time exposure effect of 1,500 ppm/min. Lungs were perfused with Krebs-Henseleit buffer at 40 ml/min from 70 to 150 min after exposure. Pulmonary artery pressure (Ppa), tracheal pressure (Pt), and the rate of lung weight gain (LWG) were measured continuously. Perfusate concentration of peptide leukotrienes LTC4, D4, and E4 were measured every 20 min during perfusion. At the conclusion of the experiment, lung tissue was analyzed for reduced and oxidized glutathione (GSH and GSSG) and lipid peroxidation (thiobarbituric acid-reactive substances, TBARS). Exposure to phosgene significantly increased Pt, LWG, LTC4, D4, and E4, TBARS, and GSSG over time compared with controls. Compared with phosgene, intratracheal NAC lowered Ppa, LWG, LTC4, D4, and E4, TBARS, and GSSG. We conclude that NAC protected against phosgene-induced lung injury by acting as an antioxidant by maintaining protective levels of glutathione, reducing both lipid peroxidation and production of arachidonic acid metabolites.
Assuntos
Acetilcisteína/uso terapêutico , Fosgênio/intoxicação , Edema Pulmonar/induzido quimicamente , Edema Pulmonar/prevenção & controle , Acetilcisteína/farmacologia , Animais , Antioxidantes/farmacologia , Glutationa/metabolismo , Leucotrienos/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Oxirredução , Coelhos , Fatores de TempoRESUMO
Exposing rabbits for 1 h to 100% O2 at 4 atm barometric pressure markedly increases the concentration of thromboxane B2 in alveolar lavage fluid [1,809 +/- 92 vs. 99 +/- 24 (SE) pg/ml, P less than 0.001], pulmonary arterial pressure (110 +/- 17 vs. 10 +/- 1 mmHg, P less than 0.001), lung weight gain (14.6 +/- 3.7 vs. 0.6 +/- 0.4 g/20 min, P less than 0.01), and transfer rates for aerosolized 99mTc-labeled diethylenetriamine pentaacetate (500 mol wt; 40 +/- 14 vs. 3 +/- 1 x 10(-3)/min, P less than 0.01) and fluorescein isothiocyanate-labeled dextran (7,000 mol wt; 10 +/- 3 vs. 1 +/- 1 x 10(-4)/min, P less than 0.01). Pretreatment with the antioxidant butylated hydroxyanisole (BHA) entirely prevents the pulmonary hypertension and lung injury. In addition, BHA blocks the increase in alveolar thromboxane B2 caused by hyperbaric O2 (10 and 45 pg/ml lavage fluid, n = 2). Combined therapy with polyethylene glycol- (PEG) conjugated superoxide dismutase (SOD) and PEG-catalase also completely eliminates the pulmonary hypertension, pulmonary edema, and increase in transfer rate for the aerosolized compounds. In contrast, combined treatment with unconjugated SOD and catalase does not reduce the pulmonary damage. Because of the striking increase in pulmonary arterial pressure to greater than 100 mmHg, we tested the hypothesis that thromboxane causes the hypertension and thus contributes to the lung injury. Indomethacin and UK 37,248-01 (4-[2-(1H-imidazol-1-yl)-ethoxy]benzoic acid hydrochloride, an inhibitor of thromboxane synthase, completely eliminate the pulmonary hypertension and edema.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Oxigenoterapia Hiperbárica/efeitos adversos , Lesão Pulmonar , Tromboxano B2/fisiologia , Animais , Líquido da Lavagem Broncoalveolar/metabolismo , Hidroxianisol Butilado/farmacologia , Hipertensão Pulmonar/etiologia , Hipertensão Pulmonar/fisiopatologia , Hipertensão Pulmonar/prevenção & controle , Imidazóis/farmacologia , Técnicas In Vitro , Indometacina/farmacologia , Pulmão/efeitos dos fármacos , Pulmão/fisiopatologia , Masculino , Edema Pulmonar/etiologia , Edema Pulmonar/fisiopatologia , Edema Pulmonar/prevenção & controle , Coelhos , Superóxido Dismutase/farmacologia , Tromboxano-A Sintase/antagonistas & inibidoresRESUMO
Vasopressor hormones alter efflux of glutathione (GSH) and increase permeability of tight junctions in perfused rat liver. Infusions of 10 nM angiotensin II, 10 microM phenylephrine, and 10 nM vasopressin significantly increased efflux of GSH into perfusate by 32-41% and decreased biliary efflux by 31-57%. Direct modulation of protein kinase C (PKC) activity by 600 nM phorbol 12,13-dibutyrate (PDB), 5 microM 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine (H-7), 5 microM sphingosine, or 10 nM staurosporine altered the pattern of efflux of GSH but not biliary oxidized glutathione disulfide (GSSG)-GSH ratios. Phorbol dibutyrate mimicked the vasopressor-mediated effects, increasing perfusate efflux by 31% and decreasing biliary efflux by 45%. Inhibitors of PKC caused qualitatively opposite responses, changing perfusate GSH by -37 to 18% and increasing biliary efflux by 22-161%. Whereas vasopressin increased penetration of [14C]sucrose into bile, modulation of PKC activity by PDB and H-7 did not affect the permeability of tight junctions to [14C]sucrose. Although pretreatment with H-7 blocked vasopressin-mediated changes in efflux of GSH, it did not prevent the increase in [14C]sucrose penetrance. We conclude that alterations in sinusoidal and biliary efflux of GSH can occur independent of changes in permeability of hepatocellular tight junctions. These findings suggest a role for protein kinase C in modulating the hepatic efflux of GSH.
Assuntos
Glutationa/metabolismo , Fígado/metabolismo , Fenilefrina/farmacologia , Proteína Quinase C/metabolismo , Vasopressinas/farmacologia , Animais , Antimetabólitos/farmacologia , Bile/metabolismo , Bile/fisiologia , Calcimicina/farmacologia , Junções Intercelulares/metabolismo , Isoxazóis/farmacologia , Masculino , Dibutirato de 12,13-Forbol/farmacologia , Proteína Quinase C/antagonistas & inibidores , Ratos , Ratos Endogâmicos , Sacarose/farmacocinéticaRESUMO
We have previously shown that phosgene markedly increases lung weight gain and pulmonary vascular permeability in rabbits. The current experiments were designed to determine whether cyclooxygenase- and lipoxygenase-derived mediators contribute to the phosgene induced lung injury. We exposed rabbits to phosgene (1,500 ppm/min), killed the animals 30 min later, and then perfused the lungs with a saline buffer for 90 min. Phosgene markedly increased lung weight gain, did not appear to increase the synthesis of cyclooxygenase metabolites, but increased 10-fold the synthesis of lipoxygenase products. Pre- or posttreatment with indomethacin decreased thromboxane and prostacyclin levels without affecting leukotriene synthesis and partially reduced the lung weight gain caused by phosgene. Methylprednisolone pretreatment completely blocked the increase in leukotriene synthesis and lung weight gain. Posttreatment with 5,8,11,14-eicosatetraynoic acid (ETYA), a nonmetabolized competitive inhibitor of arachidonic acid metabolism, or the leukotriene receptor blockers, FPL 55712 and LY 171883, also dramatically reduced the lung weight gain caused by phosgene. These results suggest that lipoxygenase products contribute to the phosgene-induced lung damage. Because phosgene exposure did not increase cyclooxygenase synthesis or pulmonary arterial pressure, we tested whether phosgene affects the lung's ability to generate or to react to thromboxane. Infusing arachidonic acid increased thromboxane synthesis to the same extent in phosgene-exposed lungs as in control lungs; however, phosgene exposure significantly reduced pulmonary vascular reactivity to thromboxane but not to angiotension II and KCl.