RESUMO
The potential toxicologic and oncogenic effects of spinosad, a natural fermentation product with insecticidal properties, were investigated. The 13-week toxicity study consisted of groups of 10 CD-1 mice/sex provided diets containing 0, 0.005, 0.015, 0.045, or 0.12% spinosad (Study 1). The 0.12% group was terminated on Test Day 44 due to mortality and overt clinical signs of toxicity. An 18-month chronic oncogenicity study consisted of groups of 50 CD-1 mice/sex provided diets containing 0, 0.0025, 0.008, or 0.036% spinosad (Study 2). Two interim groups of 10 mice/sex/group were terminated after 3 and 12 months. Females given 0.036% were terminated on Day 455 due to markedly lower body weights and feed consumption, as well as excessive mortality. Because of the early termination of the female high-dose group, additional groups of 10 male and female mice (12-month interim necrospy) and 50 male and female mice (18-month necropsy) were provided diets containing 0, 0.0008, or 0.024% spinosad (Study 3) to fully assess potential chronic toxicity and oncogenicity. Standard toxicologic parameters were evaluated consistent with existing regulatory guidelines. The primary effect in the 13-week and 18-month studies was intracellular vacuolation of histiocytic and epithelial cells in numerous tissues and organs at doses of > or = 0.015%. The histological vacuolation corresponded to ultrastructural lysosomal lamellar inclusion bodies. This alteration was consistent with phospholipidosis, a condition that results from accumulation of polar lipids in lysosomes. Lesions with no apparent direct relation to vacuolation were hyperplasia of the glandular mucosa of the stomach, skeletal muscle myopathy, bone marrow necrosis, and anemia with associated splenic hematopoiesis. The incidence of tumors in mice given spinosad was not increased relative to controls at any dose level. The no observed effect level for the 13-week study was 0.005% (6 mg/kg/day) spinosad, and for the chronic toxicity/oncogenicity study was 0.008% (11 mg/kg/day) spinosad for male and female CD-1 mice.
Assuntos
Inseticidas/toxicidade , Macrolídeos/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Testes de Carcinogenicidade , Dieta , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Ingestão de Alimentos/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/patologia , Feminino , Hepatócitos/efeitos dos fármacos , Hepatócitos/ultraestrutura , Histiócitos/efeitos dos fármacos , Histiócitos/patologia , Inseticidas/administração & dosagem , Lipidoses/induzido quimicamente , Lipidoses/patologia , Fígado/efeitos dos fármacos , Fígado/patologia , Longevidade/efeitos dos fármacos , Lisossomos/efeitos dos fármacos , Lisossomos/ultraestrutura , Macrolídeos/administração & dosagem , Masculino , Camundongos , Camundongos Endogâmicos , Necrose , Nível de Efeito Adverso não Observado , Vacúolos/efeitos dos fármacos , Vacúolos/patologiaRESUMO
Spinosad is an insecticide derived from a naturally occurring bacterium via fermentation. The toxicity of spinosad was characterized in subchronic and chronic toxicity/oncogenicity studies conducted according to standard toxicology regulatory guidelines. Subchronic toxicity was evaluated in groups of 10 Fischer 344 rats/sex given feed containing 0, 0.05, 0.1, 0.2, or 0.4% spinosad (Study 1) or 0, 0.003, 0.006, 0.012, or 0.06% spinosad (Study 2) for 13 weeks. Lower body weights and increased mortality occurred in rats given 0.4% spinosad. Microscopic effects were observed in the adrenal glands, liver, lymphoid cells, reproductive tissues, kidney, thyroid, stomach, lung, and skeletal muscle of rats given > or = 0.05% spinosad, and consisted primarily of vacuolation of cells; however, degenerative, regenerative, and/or inflammatory changes were also noted in some tissues. Vacuolation within a number of tissues was ultrastructurally characterized by an increase in size and number of lysosomes that contained extensive membranous whorls consistent with phospholipidosis. The no observed effect level (NOEL) in the 13-week studies was 0.012% (24 mg/kg/day) spinosad. Chronic toxicity and oncogenicity were evaluated in groups of 60 Fischer 344 rats/sex given feed containing 0, 0.005, 0.02, 0.05, or 0.1% spinosad for up to 2 years. Rats given 0.1% spinosad for 1 year had microscopic effects similar to those observed in the subchronic studies. Vacuolation and inflammation of the thyroid gland also occurred in rats given 0.05% spinosad for 1 year. Excessive mortality occurred in rats from the oncogenicity study given 0.1% spinosad by 21 months, and surviving rats were euthanized because the maximum tolerated dose had been exceeded. Rats given 0.05% spinosad for 2 years had vacuolation and/or inflammation involving the thyroid, lymphoid tissue, and lung. Rats given 0.05% spinosad had similar numbers of neoplasms as control rats, indicating that spinosad was not carcinogenic at dose levels up to 0.05%. The NOEL at 2 years was 0.005% (2.4 mg/kg/day) spinosad.
Assuntos
Inseticidas/toxicidade , Macrolídeos/toxicidade , Animais , Peso Corporal/efeitos dos fármacos , Testes de Carcinogenicidade , Dieta , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Feminino , Inseticidas/administração & dosagem , Lipidoses/induzido quimicamente , Lipidoses/patologia , Longevidade/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Pulmão/patologia , Tecido Linfoide/efeitos dos fármacos , Tecido Linfoide/patologia , Lisossomos/efeitos dos fármacos , Lisossomos/ultraestrutura , Macrolídeos/administração & dosagem , Masculino , Camundongos , Nível de Efeito Adverso não Observado , Tamanho do Órgão/efeitos dos fármacos , Ratos , Ratos Endogâmicos F344 , Glândula Tireoide/efeitos dos fármacos , Glândula Tireoide/patologia , Vacúolos/efeitos dos fármacos , Vacúolos/patologiaRESUMO
Phospholipidosis induced by drugs with a cationic amphiphilic structure is a generalized condition in humans and animals that is characterized by an intracellular accumulation of phospholipids and the concurrent development of concentric lamellar bodies. The primary mechanism responsible for the development of phospholipidosis is an inhibition of lysosomal phospholipase activity by the drugs. While the biochemical and ultrastructural features of the condition have been well characterized, much less effort has been directed toward understanding whether the condition has adverse effects on the organism. While there are a few cationic amphiphilic drugs that have been reported to cause phospholipidosis in humans, the principal concern with this condition is in the pharmaceutical industry during preclinical testing. While this class of drugs should technically be referred to as cationic lipophilic, the term cationic amphiphilic is widely used and recognized in this field, and for this reason, the terminology cationic amphiphilic drugs (CADs) will be employed in this Minireview. The aim of this Minireview is to provide an evaluation of the state of knowledge on the functional consequences of CAD-induced phospholipidosis.
Assuntos
Efeitos Colaterais e Reações Adversas Relacionados a Medicamentos , Lipidoses/induzido quimicamente , Fosfolipídeos/metabolismo , Animais , Humanos , Camundongos , RatosRESUMO
The pulmonary toxicity of particles is often studied using a single intratracheal instillation of the material. It was hypothesized that smaller multiple intratracheal administrations of silica would result in differences in pulmonary responses as compared to a single large intratracheal administration. In the first of a series of experiments, the pulmonary responses in male F344 rats to a single intratracheal instillation of crystalline silica (5 mg/100 g body weight) given on d 0 were compared with those resulting from 5 consecutive daily intratracheal administrations of the dust (1 mg/100 g body weight/d) with the initial dose given on d 0. Controls received saline intratracheally. In the second experiment, the dose was reduced to 1 mg/100 g body weight for the single-dose protocol and 0.2 mg/100 g body weight/d for 5 consecutive days for the multiple-dose protocol. In both experiments, responses were assessed on d 14. In the third experiment, the doses were the same as the first experiment, but the responses were assessed on d 28. The indices of toxicity were cellular differentials recovered by bronchoalveolar lavage, which is an index of inflammation, and the level of albumin in the bronchoalveolar lavage fluid, a measure of damage to the capillary-epithelial barrier. At the higher dose of silica, similar levels of inflammation and lung damage were evident in both dosing protocols. Less severe responses occurred at the lower dose. The comparative pattern between the single and multiple dosing protocols was similar in all three experiments. Since only minor differences were noted in the pulmonary responses when the responses to the single- and multiple-dose protocols were compared, data indicate that the multiple-dose protocol does not offer any advantages over the single-dose protocol.
Assuntos
Pulmão/efeitos dos fármacos , Dióxido de Silício/administração & dosagem , Dióxido de Silício/toxicidade , Animais , Biomarcadores/análise , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Relação Dose-Resposta a Droga , Esquema de Medicação , Exposição por Inalação , Instilação de Medicamentos , Pulmão/citologia , Linfócitos/citologia , Linfócitos/efeitos dos fármacos , Macrófagos Alveolares/citologia , Macrófagos Alveolares/efeitos dos fármacos , Masculino , Neutrófilos/citologia , Neutrófilos/efeitos dos fármacos , Ratos , Ratos Endogâmicos F344 , Testes de Toxicidade/métodos , TraqueiaRESUMO
Amiodarone (AD) is gaining support as a first-line antiarrhythmic drug despite its potentially fatal pulmonary toxicity involving inflammation and fibrosis. The goals of this study were to characterize a rat model of AD-induced pulmonary toxicity (AIPT) and identify a serum biomarker to aid in the diagnosis of the onset of pulmonary toxicity. Male F344 rats were instilled intratracheally with AD (6.25 mg/kg with a 3.125 mg/ml solution) in sterile water or the sterile water vehicle on days 0 and 2, a protocol that led to the development of pulmonary fibrosis on day 28 in the AD-treated animals. Animals were killed on days 3, 5, 6, 7, or 10 and bronchoalveolar lavage (BAL) was performed. Recovery of alveolar macrophages and eosinophils was increased on days 3 and 5, while neutrophil recovery and albumin levels in the first BAL fraction were significantly elevated only on day 3. BAL cells recovered from AD-treated rats at day 3 produced more phorbol myristate acetate-stimulated luminol-dependent chemiluminescence (LDCL) over 20 min than BAL cells from control rats. Experiments using specific inhibitors implicated superoxide and nitric oxide in at least part of the LDCL response. Serum levels of surfactant protein-D (SP-D), a surfactant-associated protein, were increased concurrently with the inflammatory response in the lungs. These findings indicate that this model exhibits transient pulmonary inflammation and damage, with the potential for elevated oxidant production in the lungs and subsequent pulmonary fibrosis. Also, SP-D is proposed as a specific biomarker to monitor the onset of AIPT in this model.
Assuntos
Amiodarona/toxicidade , Antiarrítmicos/toxicidade , Biomarcadores/sangue , Glicoproteínas/sangue , Pulmão/efeitos dos fármacos , Fibrose Pulmonar/induzido quimicamente , Surfactantes Pulmonares/sangue , Albuminas/metabolismo , Amiodarona/administração & dosagem , Animais , Antiarrítmicos/administração & dosagem , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Contagem de Células , Hidroxiprolina/metabolismo , Intubação Intratraqueal , Medições Luminescentes , Luminol , Pulmão/metabolismo , Pulmão/patologia , Ativação Linfocitária/efeitos dos fármacos , Masculino , Fibrose Pulmonar/sangue , Fibrose Pulmonar/patologia , Proteína D Associada a Surfactante Pulmonar , Ratos , Ratos Endogâmicos F344 , Organismos Livres de Patógenos Específicos , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
The goal of this study was to examine the relationship of the transcriptional regulatory factor nuclear factor-kappaB (NF-kappa B) to the early inflammatory events involved with silica exposure. Male F-344 rats received an intratracheal (i.t.) instillation of silica (100 mg/kg in a volume of 1 ml/kg) of saline. At 1, 3, 6, and 18 h postinstillation, and the rats were sacrificed and underwent bronchoalveolar lavage (BAL) for functional analysis of inflammation. Beginning at 1 h postinstillation, the silica-instilled (Si) rats displayed significant increases in neutrophils in BAL fluid compared to the saline controls. BAL cells from the Si group displayed a significant increase in luminol-dependent chemiluminescence (LDCL) compared to the controls. NF-kappa B activation was measurable at 3 h postinstillation, and this activation continued throughout the 18-h time course. Treatment with dexamethasone (5 mg/kg) at -3 h prior to silica instillation, at the time of instillation (0 h), and +1.5 h postinstillation resulted in both a reduction in NF-kappa B expression (by 70%) at 3 h postinstillation and corresponding reductions in LDCL, BAL cell count, and BAL neutrophils. These results show that activation of NF-kappa B is associated with silica-induced pulmonary inflammation, and the inhibition of its activation correlates temporally with suppression of inflammation.
Assuntos
Anti-Inflamatórios/farmacologia , Dexametasona/farmacologia , Pulmão/metabolismo , Pulmão/patologia , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Dióxido de Silício/toxicidade , Animais , Anti-Inflamatórios/administração & dosagem , Líquido da Lavagem Broncoalveolar/citologia , Contagem de Células/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Dexametasona/administração & dosagem , Inflamação/induzido quimicamente , Inflamação/metabolismo , Inflamação/patologia , Injeções Intraperitoneais , Intubação Intratraqueal , Medições Luminescentes , Luminol/metabolismo , Pulmão/efeitos dos fármacos , Masculino , NF-kappa B/biossíntese , Ratos , Ratos Endogâmicos F344RESUMO
Human exposure to silica (SI) is of great occupational concern because it is marked by pulmonary inflammation and fibrosis. Our objective was to determine if early pharmacological intervention altered the inflammatory and fibrotic responses to silica in rats. Male Fisher-344 rats received intratracheal (IT) instillations of the anti-inflammatory steroid, dexamethasone (DEX), incorporated into a novel liposomal (LIP) delivery system (DEX-LIP), or buffer as control (HBSS) on Day-1 and every fourth day until euthanization. On Day 0, the DEX-LIP group received IT instillations of SI (10 mg/100g body wt, DEX-LIP-SI); half of the HBSS group received SI (10 mg/100g body wt, HBSS-SI) and the other half saline (HBSS-SAL). On Day 10 or 20, bronchoalveolar lavage (BAL) was performed for cellular, biochemical, and functional analyses of inflammation and damage. HBSS-SI rats had significant elevations in the neutrophil cell count over HBSS-SAL rats at both times. DEX-LIP treatment markedly reduced these values, indicating that DEX-LIP protected against SI-induced inflammation. In contrast, DEX-LIP did not protect against biochemical (albumin concentration, and beta-glucuronidase and lactate dehydrogenase activities) and functional (luminol-dependent chemiluminescence) indices of SI-induced damage. At Day 20, the DEX-LIP treatment significantly reduced the SI-induced increase in right lung/total body weight ratio and right lung hydroxyproline content, a biochemical index of fibrosis. This attenuation of fibrosis was confirmed histopathologically on preserved left lungs from these same animals. These results show that administration of liposomes containing dexamethasone attenuated SI-induced pulmonary inflammation and fibrosis in rats, and that this protection is independent of some biochemical and functional parameters of damage.
Assuntos
Anti-Inflamatórios/administração & dosagem , Dexametasona/administração & dosagem , Lipossomos/administração & dosagem , Fibrose Pulmonar/tratamento farmacológico , Dióxido de Silício/toxicidade , Animais , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Portadores de Fármacos , Sistemas de Liberação de Medicamentos , Hidroxiprolina/análise , Intubação Intratraqueal , Contagem de Leucócitos , Pulmão/química , Pulmão/efeitos dos fármacos , Pulmão/patologia , Masculino , Neutrófilos , Tamanho do Órgão , Fibrose Pulmonar/induzido quimicamente , Fibrose Pulmonar/patologia , Ratos , Ratos Endogâmicos F344RESUMO
Previous studies have shown that vitamin E may be an integral part of lung surfactant and may function to protect this material from oxidant damage. Therefore, we measured the vitamin E levels in alveolar lavage materials from rats exposed to crystalline silica or treated with chlorphentermine (CP), two treatments that are known to increase surfactant phospholipids (PL) by different mechanisms. Silica exposure leads to increased PL synthesis, and CP treatment causes a reduction in PL degradation. Two different silica preparations, HCL-washed and unwashed silica, were used because exposure to each of them leads to different degrees of phospholipidosis. Exposure to HCL-washed silica results in a more than 17-fold increase in lavage PL and protein levels and a 12.2-fold increase in the amount of vitamin E. Exposure to unwashed silica leads to an approximately 7-fold increase in PL and proteins and a 5.8-fold increase in lavage vitamin E. Following treatment of rats with CP, there is a 15- to 19-fold increase in lavage PL and proteins and a 13.6-fold increase in vitamin E. When the results are expressed as micrograms vitamin E per milligram of lavage PL or protein, there is not much difference between controls and each treatment group. Because surfactant synthesis occurs in the endoplasmic reticulum, we also measured vitamin E in lung microsomes. Both silica exposure and CP treatment also lead to 1.8- to 2.5-fold increases, respectively, in the lung microsomal levels of vitamin E. These results demonstrate that alveolar lavage vitamin E levels are elevated along with lavage PL and proteins, and lung microsomal vitamin E levels are increased following exposure of rats to silica or treatment of the animals with CP.
Assuntos
Líquido da Lavagem Broncoalveolar/química , Clorfentermina/toxicidade , Microssomos Hepáticos/metabolismo , Fosfolipídeos/metabolismo , Dióxido de Silício/toxicidade , Simpatomiméticos/toxicidade , Vitamina E/metabolismo , Animais , Peso Corporal/efeitos dos fármacos , Clorfentermina/administração & dosagem , Injeções Intraperitoneais , Intubação Intratraqueal , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Masculino , Microssomos Hepáticos/química , Tamanho do Órgão/efeitos dos fármacos , Surfactantes Pulmonares/química , Surfactantes Pulmonares/metabolismo , Ratos , Ratos Sprague-Dawley , Dióxido de Silício/administração & dosagem , Simpatomiméticos/administração & dosagemRESUMO
The effectiveness of amiodarone in the treatment of cardiac arrhythmias is limited due to the development of pulmonary toxicity. Although the biochemical and morphologic characteristics associated with amiodarone-induced pulmonary toxicity (AIPT) are well-defined, the mechanisms underlying this disorder remain unknown. This review focuses on proposed mechanisms of AIPT, in particular (i) direct cellular damage; (ii) the role of phospholipidosis; (iii) the correlation between drug burden and toxicity; (iv) the role of the immune system; (v) the generation of oxidants; (vi) changes in membrane properties; and (vii) miscellaneous biochemical considerations. Additional discussion of the role of amiodarone's primary metabolite, desethylamiodarone, in AIPT and the involvement of preexisting lung dysfunction in the susceptibility to AIPT is included. With a clearer understanding of the possible contributions of these mechanisms to AIPT, it may be possible to develop strategies to alleviate toxicity and prolong the usefulness of amiodarone in the treatment of cardiac arrhythmias.
Assuntos
Amiodarona/efeitos adversos , Antiarrítmicos/efeitos adversos , Pulmão/efeitos dos fármacos , Amiodarona/farmacocinética , Amiodarona/toxicidade , Animais , Antiarrítmicos/farmacocinética , Antiarrítmicos/toxicidade , Membrana Celular/efeitos dos fármacos , Humanos , Sistema Imunitário/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Oxirredução , Fosfolipídeos/metabolismoRESUMO
The effect of the induction of pulmonary phospholipidosis by amiodarone on selected pulmonary host defense functions was studied in male Fischer-344 rats. One week of daily amiodarone treatment resulted in a 4.5-fold increase in total phospholipid in alveolar macrophages recovered from the lungs by bronchoalveolar lavage. The presence of the phospholipidosis had no effect on the phagocytosis of heat-killed yeast cells, the induction of luminol-dependent chemiluminescence, or the spontaneous release of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha), or spontaneous and LPS-stimulated release of IL-1 by alveolar macrophages in vitro. In contrast, the LPS-stimulated release of IL-6 and TNF-alpha by phospholipidotic alveolar macrophages was enhanced compared with control cells. The pulmonary clearance of Listeria monocytogenes following intratracheal administration of the bacteria was not affected by the phospholipidotic condition. It appears that, in the context of the functions studied, the induction of pulmonary phospholipidosis by amiodarone does not impair pulmonary host defense processes in rats, and may actually be associated with the augmentation of some activities.
Assuntos
Amiodarona/toxicidade , Antiarrítmicos/toxicidade , Imunidade Inata/efeitos dos fármacos , Lipidoses/induzido quimicamente , Pulmão/efeitos dos fármacos , Fosfolipídeos/metabolismo , Animais , Interleucinas/metabolismo , Listeriose/fisiopatologia , Medições Luminescentes , Pulmão/imunologia , Macrófagos Alveolares/efeitos dos fármacos , Macrófagos Alveolares/fisiologia , Masculino , Alvéolos Pulmonares/fisiologia , Ratos , Ratos Endogâmicos F344 , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
A previous study demonstrated that the acute phase of silica-induced lung injury in rats can be attenuated by concomitant administration of amiodarone, a cationic amphiphilic drug that inhibits phospholipase activity in the lungs. The purpose of the present study was to determine whether continued amiodarone administration could inhibit subchronic silica-induced lung injury and fibrosis. Male Fischer-344 rats were administered amiodarone (150 mg/kg, p.o., 5 days/week) for 14 days and were then instilled with silica (100 mg/kg) intratracheally. Amiodarone treatment then continued for 60 days. Injury was evaluated by parameters in bronchoalveolar lavage fluid and fibrosis was assessed by lung hydroxyproline content and trichrome staining of collagen. Within the bronchoalveolar lavage fluid, amiodarone treatment resulted in significant decreases in silica-induced elevations in albumin levels, lactate dehydrogenase activity, beta-glucuronidase activity, and neutrophil influx. Amiodarone treatment resulted in significant reductions in silica-induced increases in lung weight and hydroxyproline levels; the diminution of fibrosis due to amiodarone treatment was confirmed histologically. These results indicate that subchronic pulmonary inflammation and fibrosis induced by silica in the rat can be attenuated by the concomitant administration of amiodarone.
Assuntos
Amiodarona/uso terapêutico , Inibidores Enzimáticos/uso terapêutico , Fibrose Pulmonar/tratamento farmacológico , Silicose/tratamento farmacológico , Albuminas/análise , Albuminas/efeitos dos fármacos , Amiodarona/análogos & derivados , Amiodarona/análise , Animais , Peso Corporal/efeitos dos fármacos , Líquido da Lavagem Broncoalveolar/química , Glucuronidase/efeitos dos fármacos , Glucuronidase/metabolismo , Granuloma/patologia , Histiocitose/patologia , Hidroxiprolina/análise , Intubação Intratraqueal , L-Lactato Desidrogenase/efeitos dos fármacos , L-Lactato Desidrogenase/metabolismo , Pulmão/química , Pulmão/efeitos dos fármacos , Pulmão/patologia , Doenças Pulmonares Intersticiais/patologia , Masculino , Neutrófilos/efeitos dos fármacos , Tamanho do Órgão/efeitos dos fármacos , Fibrose Pulmonar/induzido quimicamente , Ratos , Projetos de Pesquisa , Dióxido de Silício , Silicose/patologiaRESUMO
Amiodarone (AD) and its major metabolite, desethylamiodarone (desethylAD), are both phospholipogenic. The present study was undertaken to evaluate the comparative susceptibilities of male Fischer-344 and Sprague-Dawley rats to AD-induced phospholipidosis in alveolar macrophages (AMs), liver and kidney tissue and the concomitant accumulation of AD and desethylAD in these cells, tissues and plasma. Rats were administered AD (100 mg/kg/day, p.o.) for 1 week. Plasma concentrations of AD and desethylAD were approximately 4- and 12-fold higher, respectively, in Fischer-344s compared to Sprague-Dawleys 24 h after the last dose. AD and desethylAD levels in AMs were approximately 12- and 25-fold higher, respectively, in Fischer-344s than Sprague-Dawleys. In the liver and kidney, levels of both compounds were also significantly higher in Fischer-344s than Sprague-Dawleys. Ultrastructural features indicative of phospholipidosis were not observed consistently in any tissue except AMs from treated Fischer-344s. AM total phospholipid increased nearly 5-fold in Fischer-344s, while Sprague-Dawleys showed no increase over control. AMs from both strains incubated with 10 microM AD or desethylAD in vitro were not significantly different in their accumulation of the compounds. When incubated with AD or desethylAD, the lysosomal phospholipases A1 partially purified from AMs of both strains were equally sensitive to inhibition as measured by the drug concentration giving 50% inhibition in activity (IC50). The results of this study indicate that at the same administered dose, AD and desethylAD, accumulate to higher tissue levels and are more phospholipogenic in male Fischer-344 rats than in male Sprague-Dawley rats. The basis for the high susceptibility of Fischer-344 rats to AM-induced phospholipidosis is unknown at present but appears not to be related to biochemical or cellular features of the AMs.
Assuntos
Amiodarona/metabolismo , Amiodarona/toxicidade , Antiarrítmicos/toxicidade , Lipidoses/induzido quimicamente , Fosfolipídeos/metabolismo , Amiodarona/análogos & derivados , Amiodarona/sangue , Animais , Antiarrítmicos/metabolismo , Células Cultivadas , Rim/metabolismo , Fígado/metabolismo , Macrófagos Alveolares/química , Macrófagos Alveolares/metabolismo , Macrófagos Alveolares/ultraestrutura , Masculino , Fosfolipases A/antagonistas & inibidores , Fosfolipases A/metabolismo , Fosfolipídeos/análise , Ratos , Ratos Endogâmicos F344 , Ratos Sprague-DawleyRESUMO
Exposure to silica, a cytotoxic and fibrogenic mineral dust, has been demonstrated to cause pulmonary inflammation and damage to the lung tissue. In contrast to the long-term consequences, little information exists on the sequence of inflammatory/damaging events occurring acutely after exposure to silica. The purpose of this study was to determine the minimum time after the administration of silica that the inflammatory/damage response is detectable and the temporal relationship of these processes. Male Fischer 344 rats were dosed intratracheally with silica (2.5 or 10 mg/100 g body weight) or saline vehicle. At 2 and 4 h after instillation, both cellular (total cell count and neutrophil count) and biochemical (total protein, albumin, and beta-glucuronidase and lactate dehydrogenase activities) parameters of inflammation and damage were evaluated in the bronchoalveolar lavage fluid. At 2 h, total protein levels were elevated at both silica doses, but all other parameters were unchanged; however, 4 h after silica exposure all parameters were elevated over those of the saline control. In a further attempt to characterize the inflammatory/damage processes, luminol-dependent chemiluminescence (LDCL) was performed on aliquots of chopped lung. At 2 h after silica instillation, phorbol myristate acetate-stimulated lung tissue from silica-treated rats had no increase in light production when compared to controls, whereas after 4 h there were significant increases in LDCL activity in both dose groups when compared to controls. The addition of superoxide dismutase (SOD) decreased LDCL activity of the 2.5 mg/100 g group by 59% (2 h) and 66% (4 h), and of the 10 mg/100 g group by 49% (2 h) and 73% (4 h). Alternatively, the addition of N-omega-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase, decreased the 2.5 mg/100 g group by 52% (2 h) and 60% (4 h). The 10 mg/100 g group was decreased by 67% (2 h), but only exhibited a 12% reduction at 4 h. SOD and L-NAME also inhibited the background LDCL in saline-treated rats. These reductions in LDCL activity indicate that reactive oxygen and nitrogen species play a role in the acute phase pulmonary response from silica. The results of this study indicate that the initial stages of damage begin to appear by 2 h, but damage and inflammation are definitive by 4 h after administration of silica in rats.
Assuntos
Líquido da Lavagem Broncoalveolar/química , Pulmão/efeitos dos fármacos , Dióxido de Silício/toxicidade , Albuminas/metabolismo , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Líquido da Lavagem Broncoalveolar/citologia , Contagem de Células/efeitos dos fármacos , Glucuronidase/metabolismo , L-Lactato Desidrogenase/metabolismo , Medições Luminescentes , Luminol/química , Masculino , NG-Nitroarginina Metil Éster , Neutrófilos/efeitos dos fármacos , Óxido Nítrico Sintase/antagonistas & inibidores , Proteínas/metabolismo , Ratos , Ratos Endogâmicos F344 , Dióxido de Silício/administração & dosagem , Superóxido Dismutase/farmacologia , Acetato de Tetradecanoilforbol/farmacologia , Traqueia/efeitos dos fármacosRESUMO
Intratracheal instillation of the antiarrhythmic drug amiodarone (AD) in hamsters is an established animal model of AD-induced pulmonary fibrosis. A metabolite of AD, desethylamiodarone (dAD), has also been shown to produce pulmonary fibrosis in this model. It was previously reported that following intratracheal instillation of AD in hamsters, metabolite could not be detected in lung tissue. However, in studies in our laboratory dAD was detected following instillation of AD. The goal of the present study was to monitor the distribution of AD and dAD to lung and liver within 1 h of AD instillation and to investigate the site of AD metabolism. Both AD and dAD were detected in the lung and liver within 5 min of AD instillation; lung and liver concentrations of AD and dAD were also quantified at 30 and 60 min following AD instillation. Incubation of lung and liver microsomes with AD showed that the liver is a probable site of AD metabolism following the intratracheal administration of AD to hamsters.
Assuntos
Amiodarona/análogos & derivados , Amiodarona/metabolismo , Antiarrítmicos/metabolismo , Fígado/metabolismo , Pulmão/metabolismo , Amiodarona/administração & dosagem , Animais , Antiarrítmicos/administração & dosagem , Cricetinae , Modelos Animais de Doenças , Intubação Intratraqueal , Cinética , Masculino , Mesocricetus , Microssomos/metabolismo , Fibrose Pulmonar/induzido quimicamente , Umbeliferonas/metabolismoRESUMO
Amiodarone (AD) has been shown to produce a transient pulmonary fibrosis in hamsters after intratracheal (i.t.) instillation. The goal of this study was to examine bronchoalveolar lavage (BAL) parameters during the development of fibrosis after i.t. AD in hamsters and to examine the responses to oral AD in hamsters for comparison to responses to i.t. AD in an effort to explore the roles of inflammation, phospholipidosis, and lung drug burden in AD-induced pulmonary disease. Two i.t. instillations on Days 0 and 7 of AD in hamsters produced fibrosis as characterized by elevated lung hydroxyproline content and variable increases in lavage macrophage, neutrophil, and eosinophil number through Day 28. Intratracheal AD also increased the permeability of the alveolar-capillary barrier as evidenced by an increase in BAL fluid albumin only on Day 8. Pulmonary phospholipidosis was not induced by i.t. AD and only small amounts of AD and its metabolite desethyl-AD (dAD) were detected in lung tissue through Day 10 after instillation on Days 0 and 7. The repeated oral administration of AD did not result in pulmonary fibrosis during the 35-day course of this study. Oral AD did cause a sustained increase in BAL fluid neutrophil number; other BAL cells were only slightly affected. Oral AD did not increase BAL fluid albumin content but a prominent BAL cell phospholipidosis was noted. Measurement of AD and dAD in lung tissue demonstrated a substantial accumulation of drug and metabolite after oral treatment with AD. The results of this study indicate that lung drug burden, pulmonary phospholipidosis, and lung neutrophil influx are not crucial factors in the development of AD-induced pulmonary fibrosis in hamsters. This study supports the possible involvement of physical damage to the lung and/or pulmonary eosinophilia in the generation of AD-induced pulmonary fibrosis in hamsters.
Assuntos
Amiodarona/toxicidade , Fibrose Pulmonar/induzido quimicamente , Administração Oral , Amiodarona/administração & dosagem , Animais , Líquido da Lavagem Broncoalveolar/química , Cricetinae , Hidroxiprolina/metabolismo , Intubação Intratraqueal , Masculino , Mesocricetus , Fibrose Pulmonar/metabolismoRESUMO
Chlorphentermine (CP) is a cationic, amphiphilic drug (CAD) that has been studied widely for its ability to induce phospholipidosis, a disorder characterized by excessive accumulation of cellular phospholipid and ultrastructural development of lysosomal lamellar bodies (LLBs) in the cell. The accumulation of inducing drug correlates with increasing phospholipids. In the present study, we examined the reversibility of this disorder in rat alveolar macrophages (AMs) following a 7-day treatment (30 mg/kg/day, ip). The reversibility of phospholipidosis was examined under in vivo conditions and under in vitro conditions in cell cultures for a period of up to 12 days. There was a marked reduction in cellular CP levels and phospholipid content after 4 days of recovery, both in vivo and in vitro; however, there was no indication of significant loss of LLBs. Beyond this time point, ultrastructural recovery from phospholipidosis lagged behind the biochemical recovery temporally and was somewhat less rapid in vitro than in vivo. By 12 days of recovery, AMs from both groups had recovered biochemically, but a moderate level of LLBs was still present in some AMs in the in vitro recovery group. The results of this study indicate that there are more similarities than differences when comparing the recovery of phospholipidotic cells in vitro to that occurring in vivo. We conclude that the use of cell cultures may prove valuable in studying the reversibility of CAD-induced phospholipidosis.
Assuntos
Clorfentermina/toxicidade , Lipidoses/patologia , Macrófagos Alveolares/química , Fosfolipídeos/metabolismo , Animais , Células Cultivadas , Clorfentermina/análise , Lipidoses/induzido quimicamente , Lisossomos/efeitos dos fármacos , Lisossomos/patologia , Macrófagos Alveolares/ultraestrutura , Masculino , Fosfolipídeos/análise , Ratos , Ratos Sprague-DawleyRESUMO
The use of the antiarrythmic drug amiodarone (AD) has been limited by the propensity of the drug to cause severe lung damage. AD has been shown to produce a transient pulmonary fibrosis in hamsters after intratracheal instillation. The goal of this study was to characterize the early inflammatory events associated with the administration of AD. Male Syrian hamsters that were instilled intratracheally with AD or saline vehicle underwent bronchoalveolar lavage (BAL). Total cells, macrophages, and eosinophils obtained by BAL were elevated by AD treatment at day 3. At both days 1 and 3 after instillation, AD-treated animals had significant elevations in neutrophil number. BAL fluid albumin was significantly elevated at day 1 in treated animals. Chemiluminescence (CL) performed on cells obtained by BAL showed an increase in CL of AD-treated samples compared to controls in phorbol myristate acetate (PMA) stimulated CL. PMA-induced increases in responsiveness were diminished by superoxide dismutase and catalase. These results indicate that oxidants such as superoxide and hydrogen peroxide may be involved in this inflammatory process. The results of this study show that intratracheal instillation of AD results in an inflammatory response that can be assessed by cellular, biochemical, and functional means.
Assuntos
Amiodarona , Pneumonia/induzido quimicamente , Doença Aguda , Amiodarona/administração & dosagem , Animais , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Cricetinae , Intubação Intratraqueal , Medições Luminescentes , Macrófagos/patologia , Masculino , Mesocricetus , Neutrófilos/patologia , Pneumonia/metabolismo , Pneumonia/patologia , Albumina Sérica/análise , Acetato de Tetradecanoilforbol/farmacologiaRESUMO
Chemical alteration of the glucocorticoid, methylprednisolone, has led to the introduction of a new class of compounds called the 21-aminosteroids (21-ASs). The purpose of this study was to investigate the effect of the 21-AS, U74389G, on silica-induced acute lung injury. Male Fischer 344 rats were treated intraperitoneally with saline or U74389G in a total dose of 15 mg/kg divided into three injections of 5 mg/kg separated by 4 h. Following the first treatment, animals from the two groups were intratracheally instilled with silica (10 mg/100 g body wt in 0.5 ml of saline) or saline vehicle (0.5 ml). Twenty-four hours after the instillations, bronchoalveolar lavage (BAL) was performed. In the animals not receiving U74389G, marked increases in total protein, beta-glucuronidase, and lactate dehydrogenase (LDH) activities and number of neutrophils (PMNs) were demonstrated in the BAL fluid of the silica-treated animals compared to their controls. Silica also caused dramatic increases in the luminol-dependent chemiluminescence (CL) of lung tissue and BAL cells. The CL reaction was decreased by superoxide dismutase (SOD) and N-nitro-L-arginine methyl ester hydrochloride (L-NAME), a nitric oxide (NO) synthase inhibitor. In animals treated with U74389G, there was attenuation of the silica-induced increases in biochemical, cellular, and chemiluminescent indices of damage. This study demonstrates that U74389G significantly reduces acute lung injury caused by the intratracheal instillation of silica, and this drug may be of potential value for treatment of lung diseases in which damage caused by reactive oxygen species has been implicated.
Assuntos
Pulmão/efeitos dos fármacos , Pneumonia/induzido quimicamente , Pneumonia/prevenção & controle , Pregnatrienos/farmacologia , Dióxido de Silício/farmacologia , Doença Aguda , Animais , Arginina/análogos & derivados , Arginina/farmacologia , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Glucuronidase/metabolismo , L-Lactato Desidrogenase/metabolismo , Medições Luminescentes , Luminol , Masculino , NG-Nitroarginina Metil Éster , Neutrófilos/patologia , Óxido Nítrico/antagonistas & inibidores , Proteínas/metabolismo , Ratos , Ratos Endogâmicos F344 , Superóxido Dismutase/farmacologiaRESUMO
The inhalation of silica has been shown to produce a dramatic inflammatory and toxic response within the lungs of humans and laboratory animals. A variety of cellular and biochemical parameters are used to assess the silica-induced lung injury. The purpose of this paper is to introduce the use of luminol-dependent chemiluminescence as a new method to study inflammation in both phagocytic cells and lung tissue recovered from silica-exposed animals. Chemiluminescence, or the emission of light, accompanies the release of reactive forms of oxygen when phagocytic cells are challenged. In this study, male Fischer 344 rats were intratracheally instilled with either silica (10 mg/100 g bw) or saline vehicle. One day after the instillations, a marked increase in the chemiluminescence was observed in the lung tissue and bronchoalveolar lavage cells recovered from the silica-treated animals when compared with the saline controls. The light reaction was markedly decreased by either superoxide dismutase of N-nitro-arginine methyl ester hydrochloride. Superoxide dismutase is involved in the enzymatic breakdown of superoxide anion, while N-nitro-L-arginine methyl ester hydrocholoride, a nitric oxide synthase inhibitor, prevents the formation of nitric oxide. When superoxide anion and nitric oxide react, they form the highly oxidizing substance peroxynitrite. This study then implicates peroxynitrite as an agent that may be responsible for some of the oxidant lung injury that is associated with silica exposure. The use of luminol-dependent chemiluminescence may prove valuable as a method to measure the earliest events in the inflammatory process, and may be an adjunct in studying the mechanisms that produce inflammation.