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1.
Chem Res Toxicol ; 32(9): 1737-1747, 2019 09 16.
Artigo em Inglês | MEDLINE | ID: mdl-31407890

RESUMO

The biological response of bronchial epithelial cells to particles is associated with a sequestration of cell metal by the particle surface and a subsequent disruption in host iron homeostasis. The macrophage is the cell type resident in the respiratory tract that is most likely to make initial contact with inhaled particles. We tested the postulates that (1) silica, a prototypical particle, disrupts iron homeostasis in alveolar macrophages (AMs); and (2) the altered iron homeostasis results in both an oxidative stress and pro-inflammatory effects. Human AMs (1.0 × 106/mL) demonstrated an increased import of iron following particle exposure with nonheme iron concentrations of 0.57 ± 0.03, 1.72 ± 0.09, 0.88 ± 0.09, and 3.21 ± 0.11 ppm in cells exposed for 4 h to media, 500 µM ferric ammonium citrate (FAC), 100 µg/mL silica, and both silica and FAC, respectively. Intracellular ferritin concentrations and iron release were similarly increased after AM exposure to FAC and silica. Silica increased oxidant generation by AMs measured using both dichlorofluorescein diacetate fluorescence and reduction of nitroblue tetrazolium salt. Concentrations of interleukin (IL)-1ß, IL-6, IL-8, and tumor necrosis factor-α in macrophage supernatant increased following 100 µg/mL silica exposure for 24 h. Treatment of AMs with 500 µM FAC decreased both oxidant generation and cytokine release associated with silica exposure, supporting a dependence of these effects on sequestration of cell metal by the particle surface. We conclude that (1) silica exposure disrupts iron homeostasis resulting in increased import, accumulation, and release of the metal; and (2) the altered iron homeostasis following silica exposure impacts oxidant generation and pro-inflammatory effects.


Assuntos
Homeostase/efeitos dos fármacos , Inflamação/induzido quimicamente , Ferro/metabolismo , Macrófagos Alveolares/efeitos dos fármacos , Quartzo/toxicidade , Acetofenonas/farmacologia , Animais , Linhagem Celular Tumoral , Citocinas/metabolismo , Inibidores Enzimáticos/farmacologia , Compostos Férricos/farmacologia , Ferritinas/metabolismo , Humanos , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , NADPH Oxidase 2/genética , NADPH Oxidases/antagonistas & inibidores , Estresse Oxidativo/efeitos dos fármacos , Compostos de Amônio Quaternário/farmacologia
2.
Part Fibre Toxicol ; 10: 25, 2013 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-23800224

RESUMO

BACKGROUND: We tested the hypothesis that normal human bronchial epithelial (NHBE) cells 1) grown submerged in media and 2) allowed to differentiate at air-liquid interface (ALI) demonstrate disparities in the response to particle exposure. RESULTS: Following exposure of submerged NHBE cells to ambient air pollution particle collected in Chapel Hill, NC, RNA for IL-8, IL-6, heme oxygenase 1 (HOX1) and cyclooxygenase 2 (COX2) increased. The same cells allowed to differentiate over 3, 10, and 21 days at ALI demonstrated no such changes following particle exposure. Similarly, BEAS-2B cells grown submerged in media demonstrated a significant increase in IL-8 and HOX1 RNA after exposure to NIST 1648 particle relative to the same cells exposed after growth at ALI. Subsequently, it was not possible to attribute the observed decreases in the response of NHBE cells to differentiation alone since BEAS-2B cells, which do not differentiate, showed similar changes when grown at ALI. With no exposure to particles, differentiation of NHBE cells at ALI over 3 to 21 days demonstrated significant decrements in baseline levels of RNA for the same proteins (i.e. IL-8, IL-6, HOX1, and COX2). With no exposure to particles, BEAS-2B cells grown at ALI showed comparable changes in RNA for IL-8 and HOX1. After the same particle exposure, NHBE cells grown at ALI on a transwell in 95% N2-5% CO2 and exposed to NIST 1648 particle demonstrated significantly greater changes in IL-8 and HOX1 relative to cells grown in 95% air-5% CO2. CONCLUSIONS: We conclude that growth of NHBE cells at ALI is associated with a diminished biological effect following particle exposure relative to cells submerged in media. This decreased response showed an association with increased oxygen availability.


Assuntos
Brônquios/efeitos dos fármacos , Diferenciação Celular , Proliferação de Células , Células Epiteliais/efeitos dos fármacos , Heme Oxigenase-1/metabolismo , Material Particulado/toxicidade , Brônquios/metabolismo , Técnicas de Cultura de Células , Hipóxia Celular , Linhagem Celular , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Células Epiteliais/metabolismo , Regulação da Expressão Gênica , Heme Oxigenase-1/genética , Humanos , Interleucina-6/genética , Interleucina-6/metabolismo , Interleucina-8/genética , Interleucina-8/metabolismo , Oxigênio/metabolismo , RNA Mensageiro/metabolismo , Fatores de Tempo
3.
Biometals ; 23(4): 657-67, 2010 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-20336479

RESUMO

Exposure to bleomycin can result in an inflammatory lung injury. The biological effect of this anti-neoplastic agent is dependent on its coordination of iron with subsequent oxidant generation. In lung cells, divalent metal transporter 1 (DMT1) can participate in metal transport resulting in control of an oxidative stress and tissue damage. We tested the postulate that metal import by DMT1 would participate in preventing lung injury after exposure to bleomycin. Microcytic anemia (mk/mk) mice defective in DMT1 and wild-type mice were exposed to either bleomycin or saline via intratracheal instillation and the resultant lung injury was compared. Twenty-four h after instillation, the number of neutrophils and protein concentrations after bleomycin exposure were significantly elevated in the mk/mk mice relative to the wild-type mice. Similarly, levels of a pro-inflammatory mediator were significantly increased in the mk/mk mice relative to wild-type mice following bleomycin instillation. Relative to wild-type mice, mk/mk mice demonstrated lower non-heme iron concentrations in the lung, liver, spleen, and splenic, peritoneal, and liver macrophages. In contrast, levels of this metal were elevated in alveolar macrophages from mk/mk mice. We conclude that DMT1 participates in the inflammatory lung injury after bleomycin with mk/mk mice having increased inflammation and damage following exposure. This finding supports the hypothesis that DMT1 takes part in iron detoxification and homeostasis in the lung.


Assuntos
Bleomicina/farmacologia , Proteínas de Transporte de Cátions/deficiência , Lesão Pulmonar/induzido quimicamente , Lesão Pulmonar/metabolismo , Anemia/genética , Anemia/metabolismo , Animais , Proteínas de Transporte de Cátions/genética , Feminino , Ferritinas/metabolismo , Homeostase , Pulmão/citologia , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Macrófagos Alveolares/citologia , Macrófagos Alveolares/metabolismo , Masculino , Metais/metabolismo , Camundongos , Camundongos Knockout , Baço/citologia , Baço/metabolismo
4.
Inhal Toxicol ; 22(2): 169-78, 2010 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19757987

RESUMO

In situ reactions of metal ions or their compounds are important mechanisms by which particles alter lung immune responses. The authors hypothesized that major determinants of the immunomodulatory effect of any metal include its redox behavior/properties, oxidation state, and/or solubility, and that the toxicities arising from differences in physicochemical parameters are manifest, in part, via differential shifts in lung iron (Fe) homeostasis. To test the hypotheses, immunomodulatory potentials for both pentavalent vanadium (VV; as soluble metavanadate or insoluble vanadium pentoxide) and hexavalent chromium (CrVI; as soluble sodium chromate or insoluble calcium chromate) were quantified in rats after inhalation (5h/day for 5 days) of each at 100 microg metal/m3. Differences in effects on local bacterial resistance between the two VV, and between each CrVI, agents suggested that solubility might be a determinant of in situ immunotoxicity. For the soluble forms, VV had a greater impact on resistance than CrVI, indicating that redox behavior/properties was likely also a determinant. The soluble VV agent was the strongest immunomodulant. Regarding Fe homeostasis, both VV agents had dramatic effects on airway Fe levels. Both also impacted local immune/airway epithelial cell Fe levels in that there were significant increases in production of select cytokines/chemokines whose genes are subject to regulation by HIF-1 (whose intracellular longevity is related to cell Fe status). Our findings contribute to a better understanding of the role that metal compound properties play in respiratory disease pathogenesis and provide a rationale for differing pulmonary immunotoxicities of commonly encountered ambient metal pollutants.


Assuntos
Antibacterianos , Cromo/farmacologia , Cromo/toxicidade , Homeostase/efeitos dos fármacos , Ferro/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/microbiologia , Vanádio/farmacologia , Vanádio/toxicidade , Animais , Câmaras de Exposição Atmosférica , Carga Corporal (Radioterapia) , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Quimiocina CCL2/metabolismo , Cromo/química , Ferritinas/metabolismo , Imunidade/efeitos dos fármacos , Proteínas de Ligação ao Ferro/metabolismo , Listeria monocytogenes/efeitos dos fármacos , Pulmão/imunologia , Masculino , Ratos , Ratos Endogâmicos F344 , Transferrina/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Vanádio/química
5.
Biometals ; 22(5): 803-15, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19306086

RESUMO

Despite recurrent exposure to zinc through inhalation of ambient air pollution particles, relatively little information is known about the homeostasis of this metal in respiratory epithelial cells. We describe zinc uptake and release by respiratory epithelial cells and test the postulate that Zn(2+) transport interacts with iron homeostasis in these same cells. Zn(2+) uptake after 4 and 8 h of exposure to zinc sulfate was concentration- and time-dependent. A majority of Zn(2+) release occurred in the 4 h immediately following cell exposure to ZnSO(4). Regarding metal importers, mRNA for Zip1 and Zip2 showed no change after respiratory epithelial cell exposure to zinc while mRNA for divalent metal transporter (DMT)1 increased. Western blot assay for DMT1 protein supported an elevated expression of this transport protein following zinc exposure. RT-PCR confirmed mRNA for the metal exporters ZnT1 and ZnT4 with the former increasing after ZnSO(4). Cell concentrations of ferritin increased with zinc exposure while oxidative stress, measured as lipid peroxides, was decreased supporting an anti-oxidant function for Zn(2+). Increased DMT1 expression, following pre-incubations of respiratory epithelial cells with TNF-alpha, IFN-gamma, and endotoxin, was associated with significantly decreased intracellular zinc transport. Finally, incubations of respiratory epithelial cells with both zinc sulfate and ferric ammonium citrate resulted in elevated intracellular concentrations of both metals. We conclude that exposure to zinc increases iron uptake by respiratory epithelial cells. Elevations in cell iron can possibly affect an increased expression of DMT1 and ferritin which function to diminish oxidative stress. Comparable to other metal exposures, changes in iron homeostasis may contribute to the biological effects of zinc in specific cells and tissues.


Assuntos
Poluentes Atmosféricos/metabolismo , Brônquios/citologia , Células Epiteliais/metabolismo , Homeostase/fisiologia , Ferro/metabolismo , Zinco/metabolismo , Poluentes Atmosféricos/toxicidade , Transporte Biológico , Western Blotting , Linhagem Celular , Células Cultivadas , Células Epiteliais/efeitos dos fármacos , Humanos , Estresse Oxidativo/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Zinco/toxicidade
6.
Am J Respir Crit Care Med ; 178(11): 1130-8, 2008 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-18723436

RESUMO

RATIONALE: Lung injury after cigarette smoking is related to particle retention. Iron accumulates with the deposition of these particles. OBJECTIVES: We tested the postulate that (1) injury after smoking correlates with exposure to the particulate fraction of cigarette smoke, (2) these particles alter iron homeostasis, triggering metal accumulation, and (3) this alteration in iron homeostasis affects oxidative stress and inflammation. METHODS: Rats and human respiratory epithelial cells were exposed to cigarette smoke, filtered cigarette smoke, and cigarette smoke condensate (the particulate fraction of smoke), and indices of iron homeostasis, oxidative stress, and inflammatory injury were determined. Comparable measures were also evaluated in nonsmokers and smokers. MEASUREMENTS AND MAIN RESULTS: After exposure of rats to cigarette smoke, increased lavage concentrations of iron and ferritin, serum ferritin levels, and nonheme iron concentrations in the lung and liver tissue all increased. Lavage ascorbate concentrations were decreased, supporting an oxidative stress. After filtering of the cigarette smoke to remove particles, most of these changes were reversed. Exposure of cultured respiratory epithelial cells to cigarette smoke condensate caused a similar accumulation of iron, metal-dependent oxidative stress, and increased IL-8 release. Lavage samples in healthy smokers and smoking patients with chronic obstructive pulmonary disease revealed elevated concentrations of both iron and ferritin relative to healthy nonsmokers. Lavage ascorbate decreased with cigarette smoking. Serum iron and ferritin levels among smokers were increased, supporting systemic accumulation of this metal after cigarette smoke exposure. CONCLUSIONS: We conclude that cigarette smoke particles alter iron homeostasis, both in the lung and systemically.


Assuntos
Ferro/metabolismo , Lesão Pulmonar/etiologia , Lesão Pulmonar/metabolismo , Material Particulado/efeitos adversos , Fumar/efeitos adversos , Adolescente , Adulto , Animais , Líquido da Lavagem Broncoalveolar/química , Estudos de Casos e Controles , Modelos Animais de Doenças , Feminino , Homeostase , Humanos , Inflamação/etiologia , Lesão Pulmonar/complicações , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo , Ratos , Ratos Wistar , Fatores Sexuais , Adulto Jovem
7.
Am J Respir Cell Mol Biol ; 38(6): 715-23, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18203974

RESUMO

The dissociation of iron from heme is a major factor in iron metabolism and the cellular concentrations of the metal correlate with heme degradation. We tested the hypotheses that (1) exposure to a product of heme catabolism, carbon monoxide (CO), alters iron homeostasis in the lung and in cultured respiratory epithelial cells; (2) this response includes both decreased uptake and increased release of cell metal; and (3) the effects of CO on cell function track changes in metal homeostasis. In rats exposed to 50 ppm CO for 24 hours, non-heme iron concentrations decreased in the lung and increased in the liver. In respiratory epithelial cells cultured at air-liquid interface, CO exposure decreased cell non-heme iron and ferritin concentrations within 2 hours and the effect was fully reversible. CO significantly depressed iron uptake by epithelial cells, despite increased expression of divalent metal transporter-1, while iron release was elevated. The loss of non-heme iron after CO reduced cellular oxidative stress, blocked the release of the proinflammatory mediator (interleukin-8), and interfered with cell cycle protein expression. We conclude that CO reduces the iron content of the lung through both the metal uptake and release mechanisms. This loss of cellular iron after CO is in line with certain biological effects of the gas that have been implicated in the protection of cell viability.


Assuntos
Antimetabólitos/farmacologia , Monóxido de Carbono/farmacologia , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Homeostase , Ferro/metabolismo , Mucosa Respiratória , Acetaldeído/metabolismo , Animais , Antimetabólitos/metabolismo , Antioxidantes/metabolismo , Lavagem Broncoalveolar , Monóxido de Carbono/metabolismo , Linhagem Celular , Proliferação de Células , Criança , Desferroxamina/metabolismo , Células Epiteliais/citologia , Ferritinas/metabolismo , Heme/química , Heme/metabolismo , Humanos , Interleucina-8/metabolismo , Masculino , Estresse Oxidativo , Ratos , Ratos Sprague-Dawley , Mucosa Respiratória/citologia , Mucosa Respiratória/efeitos dos fármacos , Sideróforos/metabolismo
8.
Antioxid Redox Signal ; 10(2): 371-7, 2008 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17999626

RESUMO

Human exposure to asbestos can cause a wide variety of pulmonary diseases, including pneumoconiosis (i.e., asbestosis). This lung injury is mediated by oxidant generation which increases with the concentration of iron associated with the asbestos. Iron from host sources is complexed by the surface of these fibrous silicates following introduction into the lower respiratory tract. Using bronchoalveolar lavage from unexposed and exposed workers, we demonstrate that asbestos disrupts the normal iron homeostasis in the lungs. Based on these findings, we propose a model of oxidative stress and human lung injury after asbestos exposure.


Assuntos
Amianto/toxicidade , Ferro/metabolismo , Pneumopatias/induzido quimicamente , Pulmão/metabolismo , Exposição Ambiental , Ferritinas/metabolismo , Homeostase , Humanos , Lactoferrina/metabolismo , Pulmão/efeitos dos fármacos , Pulmão/patologia , Pneumopatias/metabolismo , Pneumopatias/patologia , Modelos Biológicos , Oxidantes/metabolismo , Oxidantes/toxicidade , Transferrina/metabolismo
9.
Respir Res ; 9: 10, 2008 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-18215276

RESUMO

BACKGROUND: Lung injury caused by both inhaled dusts and infectious agents depends on increased availability of iron and metal-catalyzed oxidative stress. Because inhaled particles, such as silica, and certain infections can cause secondary pulmonary alveolar proteinosis (PAP), we tested the hypothesis that idiopathic PAP is associated with an altered iron homeostasis in the human lung. METHODS: Healthy volunteers (n = 20) and patients with idiopathic PAP (n = 20) underwent bronchoalveolar lavage and measurements were made of total protein, iron, tranferrin, transferrin receptor, lactoferrin, and ferritin. Histochemical staining for iron and ferritin was done in the cell pellets from control subjects and PAP patients, and in lung specimens of patients without cardiopulmonary disease and with PAP. Lavage concentrations of urate, glutathione, and ascorbate were also measured as indices of oxidative stress. RESULTS: Lavage concentrations of iron, transferrin, transferrin receptor, lactoferrin, and ferritin were significantly elevated in PAP patients relative to healthy volunteers. The cells of PAP patients had accumulated significant iron and ferritin, as well as considerable amounts of extracellular ferritin. Immunohistochemistry for ferritin in lung tissue revealed comparable amounts of this metal-storage protein in the lower respiratory tract of PAP patients both intracellularly and extracellularly. Lavage concentrations of ascorbate, glutathione, and urate were significantly lower in the lavage fluid of the PAP patients. CONCLUSION: Iron homeostasis is altered in the lungs of patients with idiopathic PAP, as large amounts of catalytically-active iron and low molecular weight anti-oxidant depletion are present. These findings suggest a metal-catalyzed oxidative stress in the maintenance of this disease.


Assuntos
Homeostase , Ferro/metabolismo , Estresse Oxidativo , Proteinose Alveolar Pulmonar/metabolismo , Líquido da Lavagem Broncoalveolar/química , Estudos de Casos e Controles , Ferritinas/metabolismo , Humanos , Lactoferrina/metabolismo , Pulmão/metabolismo , Proteínas/metabolismo , Receptores da Transferrina/metabolismo , Transferrina/metabolismo
11.
J Inorg Biochem ; 147: 126-33, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25843360

RESUMO

Mechanistic pathways underlying inflammatory injury following exposures to vanadium-containing compounds are not defined. We tested the postulate that the in vitro biological effect of vanadium results from its impact on iron homeostasis. Human bronchial epithelial (HBE) cells exposed to vanadyl sulfate (VOSO4) showed a time- and dose-dependent increase in vanadium relative to PBS. HBE cells exposed to VOSO4 and then exposed to ferric ammonium citrate (FAC) significantly increased intracellular iron import supporting an interaction between the two metals. Following exposure to VOSO4, there was an increase (336±73%) in RNA for divalent metal transporter 1 (DMT1), a major iron importer. With inclusion of VOSO4 in the incubation, vanadium could be measured in the nuclear and mitochondrial fractions and the supernatant. Non-heme iron in the nuclear and mitochondrial fractions were decreased immediately following VOSO4 exposure while there was an increased concentration of non-heme iron in the supernatant. Provision of excess iron inhibited changes in the concentration of this metal provoked by VOSO4 exposures. Using Amplex Red, VOSO4 was shown to significantly increase oxidant generation by HBE cells in a time- and dose-dependent manner. HBE cells pre-treated with FAC and then exposed to VOSO4 demonstrated a decreased generation of oxidants. Similarly, activation of the transcription factor NF-ĸB promoter and release of interleukin-6 and -8 were increased following VOSO4 exposure and these effects were diminished by pre-treatment with FAC. We conclude that an initiating event in biological effect after exposure to vanadyl sulfate is a loss of requisite cell iron.


Assuntos
Células Epiteliais/efeitos dos fármacos , Compostos Férricos/farmacologia , Compostos de Amônio Quaternário/farmacologia , Compostos de Vanádio/farmacologia , Células Cultivadas , Humanos
12.
Hum Pathol ; 34(8): 737-42, 2003 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-14506632

RESUMO

We report on a deposition of oxalate crystals on ferruginous bodies after occupational exposure to asbestos demonstrated in 3 patients. We investigated the mechanism and possible significance of this deposition by testing the hypothesis that oxalate generated through nonenzymatic oxidation of ascorbate by asbestos-associated iron accounts for the deposition of the crystal on a ferruginous body. Crocidolite asbestos (1000 microg/mL) was incubated with 500 micromol H(2)O(2) and 500 micromol ascorbate for 24 hours at 22 degrees C. The dependence of oxalate generation on iron-catalyzed oxidant production was tested with the both the metal chelator deferoxamine and the radical scavenger dimethylthiourea. Incubation of crocidolite, H(2)O(2), and ascorbate in vitro generated approximately 42 nmol of oxalate in 24 hours. Oxalate generation was diminished significantly by the inclusion of either deferoxamine or dimethylthiourea in the reaction mixture. Incubation of asbestos bodies and uncoated fibers isolated from human lung with 500 micromol H(2)O(2) and 500 micromol ascorbate for 24 hours at 22 degrees C resulted in the generation of numerous oxalate crystals. We conclude that iron-catalyzed production of oxalate from ascorbate can account for the deposition of this crystal on ferruginous bodies.


Assuntos
Asbesto Crocidolita/metabolismo , Asbestose/metabolismo , Oxalato de Cálcio/metabolismo , Pulmão/metabolismo , Tioureia/análogos & derivados , Asbesto Crocidolita/efeitos adversos , Asbesto Crocidolita/química , Asbestose/etiologia , Asbestose/patologia , Ácido Ascórbico/química , Oxalato de Cálcio/análise , Oxalato de Cálcio/química , Cristalização , Cristalografia por Raios X , Desferroxamina/química , Evolução Fatal , Humanos , Peróxido de Hidrogênio/química , Ferro/química , Quelantes de Ferro/química , Pulmão/patologia , Masculino , Pessoa de Meia-Idade , Oxirredução , Tioureia/química
13.
J Toxicol Environ Health A ; 66(24): 2281-97, 2003 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-14686339

RESUMO

Epidemiologic evidence suggests that increased morbidity and mortality are associated with the concentrations of ambient air particulate matter (PM). Many sources contribute to the particulate fraction of ambient air pollution, including diesel exhaust particulates (DEP). Diesel exhaust also contributes gas-phase pollutants to the atmosphere, and gaseous copollutants may influence the toxicity of PM. The composition of diesel exhaust varies greatly depending on the engine load conditions as well as other factors. To determine whether different diesel exhaust composition can affect lung cell resposes, the effects of of diesel exhaust extracts derived from different engine loads were examined on normal human bronchial epithelial cells (NHBE) in vitro. Diesel exhaust was collected into chilled impingers containing phosphate-buffered saline (PBS). Cultured NHBE cells were treated with 0 to 500 microg/well extract from approximately 0% engine load (termed low load or LL) or extract from approximately 75% engine load (termed high load or HL) for 24 h. The HL extract was cytotoxic at 500 microg compared to controls as measured by (51)Cr release. Production of the neutrophil chemotaxin interleukin 8 (IL-8) was decreased 4.7-fold in cells treated with 500 microg LL extract, whereas cells treated with 500 microg HL extract showed a 2.4-fold increase in IL-8 release. Production of the inflammatory and immune system mediator prostaglandin E(2) (PGE(2)) was increased up to 2.5-fold in cells treated with HL extract, but unchanged with other treatments. Melittin stimulation of cells showed that the LL extract had an inhibitory effect on PGE(2) release at 500 microg. Differences in carbonyl content of the extracts were found by high performance liquid chromatography/mass spectroscopy HPLC/MS, with the HL extract having more intermediate size carbonyls (i.e. with six to nine carbons). The data suggest that the response of NHBE cells to treatment with diesel exhaust will vary depending on the constituent components of the exhaust.


Assuntos
Poluentes Atmosféricos/toxicidade , Brônquios/efeitos dos fármacos , Doenças Respiratórias/induzido quimicamente , Emissões de Veículos/toxicidade , Poluentes Atmosféricos/química , Poluição do Ar , Brônquios/citologia , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Células Epiteliais/efeitos dos fármacos , Humanos
14.
J Immunotoxicol ; 4(1): 49-60, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18958712

RESUMO

The in situ reactions of metal ions/complexes are important in understanding the mechanisms by which environmental and occupational metal particles alter lung immune responses. A better understanding of these reactions in situ will also allow for the improved specificity and controlled toxicity of novel metallocompounds to be used as inhaled diagnostics or therapeutics. Our previous work showed that inhalation of metals (e.g., chromium, vanadium, nickel) caused altered lung immune cell function and host resistance. The data also suggested that the degree of immunomodulation induced depended not only on the amount of metal deposited, but also the compound used. If specificity governs pulmonary immunomodulatory potential, it follows that physicochemical properties inherent to the metal have a role in the elicited effects. We hypothe-size that major determinants of any metal compound's potential are its redox behavior, valency (generally referred to as oxidation state and considered speciation in chemical literature), and/or solubility. In accord with the extensive work carried out with vanadium (chemical symbol V) compounds showing the importance of form used, differences in potential for a range of V agents (pentavalent [V(V)] insoluble vanadium pentoxide and soluble sodium metavanadate, tetravalent [V(IV)] vanadyl dipicolinate, and trivalent [V(III)] bis(dipicolinato)vanadium) were quantified based on induced changes in local bacterial resistance after host inhalation of each agent at 100 mu g V/m(3) (5 hr/d for 5 d). Differences in effect between V(V) forms indicated that solubility was a critical property in in situ pulmonary immunotoxicity. Among the soluble forms, oxidizing vanadate had the greatest impact on resistance; reducing V(III) altered resistance to a lesser extent. Both the V(IV) and insoluble V(V) had no effect. When data was analyzed in the context of pre-infection lung V burdens, soluble V agents with different oxidation states induced varying responses, supporting the hypothesis that differences in immunomodulatory potential might be attributed to redox behavior or valency. Our findings both provide a basis for understanding why some metals could be a greater health risk than others (when encountered in equal amounts) and will assist in the design of inhalable metallopharmaceuticals by allowing researchers to preempt selection of certain metal ions or complexes for use in such products.

15.
Am J Physiol Lung Cell Mol Physiol ; 292(1): L134-43, 2007 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16905637

RESUMO

We tested the hypothesis that oxidative stress and biological effect after ozone (O3) exposure are dependent on changes in iron homeostasis. After O3 exposure, healthy volunteers demonstrated increased lavage concentrations of iron, transferrin, lactoferrin, and ferritin. In normal rats, alterations of iron metabolism after O3 exposure were immediate and preceded the inflammatory influx. To test for participation of this disruption in iron homeostasis in lung injury following O3 inhalation, we exposed Belgrade rats, which are functionally deficient in divalent metal transporter 1 (DMT1) as a means of iron uptake, and controls to O3. Iron homeostasis was disrupted to a greater extent and the extent of injury was greater in Belgrade rats than in control rats. Nonheme iron and ferritin concentrations were higher in human bronchial epithelial (HBE) cells exposed to O3 than in HBE cells exposed to filtered air. Aldehyde generation and IL-8 release by the HBE cells was also elevated following O3 exposure. Human embryonic kidney (HEK 293) cells with elevated expression of a DMT1 construct were exposed to filtered air and O3. With exposure to O3, elevated DMT1 expression diminished oxidative stress (i.e., aldehyde generation) and IL-8 release. We conclude that iron participates critically in the oxidative stress and biological effects after O3 exposure.


Assuntos
Ferro/metabolismo , Lesão Pulmonar , Pulmão/efeitos dos fármacos , Ozônio/toxicidade , Adolescente , Adulto , Animais , Líquido da Lavagem Broncoalveolar/química , Proteínas de Transporte de Cátions/deficiência , Linhagem Celular , Modelos Animais de Doenças , Ferritinas/metabolismo , Homeostase , Humanos , Lactoferrina/metabolismo , Pulmão/metabolismo , Pulmão/patologia , Masculino , Estresse Oxidativo , Ozônio/administração & dosagem , Ratos , Ratos Endogâmicos F344 , Ratos Mutantes , Ratos Sprague-Dawley , Receptores da Transferrina/metabolismo , Fatores de Tempo , Transferrina/metabolismo
16.
Toxicol Pathol ; 34(6): 723-9, 2006.
Artigo em Inglês | MEDLINE | ID: mdl-17074740

RESUMO

Exposure to synthetic fibers with employment in textile mills can be associated with an elevated risk of interstitial lung disease (ILD). A mechanism of injury has not been determined. ILD can follow exposures to inorganic fibers (e.g., asbestos) which are associated with a mobilization of iron and catalysis of an oxidative stress. We describe 2 patients with ILD associated with exposure to synthetic textile fibers who demonstrated carbon-based ferruginous bodies suggesting an in vivo accumulation of iron by synthetic fibers after deposition in the lung. These iron-laden bodies varied from perfectly linear fibers to almost particulate matter. Linear structures were irregularly interrupted by deposition of iron-abundant material. The capacity of these synthetic fibers to complex iron and generate an oxidative stress is confirmed in vitro.


Assuntos
Exposição por Inalação , Pulmão/efeitos dos fármacos , Doenças Profissionais/etiologia , Exposição Ocupacional , Fibrose Pulmonar/etiologia , Indústria Têxtil , Têxteis/efeitos adversos , Adulto , Celulose/efeitos adversos , Celulose/química , Feminino , Humanos , Ferro/análise , Pulmão/química , Pulmão/metabolismo , Pulmão/patologia , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Nylons/efeitos adversos , Nylons/química , Doenças Profissionais/metabolismo , Doenças Profissionais/patologia , Doenças Profissionais/fisiopatologia , Oxidantes/química , Estresse Oxidativo/efeitos dos fármacos , Tamanho da Partícula , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/patologia , Fibrose Pulmonar/fisiopatologia , Testes de Função Respiratória , Têxteis/análise , Tomografia Computadorizada por Raios X
17.
Am J Respir Cell Mol Biol ; 34(3): 286-92, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16272461

RESUMO

Lung injury after asbestos exposure is associated with an oxidative stress that is catalyzed by iron in the fiber matrix, complexed to the surface, or both. We tested the hypothesis that the cellular response to asbestos includes the transport and sequestration of this iron through (1) generation of superoxide for ferrireduction, (2) up-regulation of divalent metal transporter-1 (DMT1) for intracellular transport of Fe2+, and (3) increased production of cellular ferritin where the metal is stored in a catalytically less reactive state. BEAS-2B cells with normal and elevated Cu,Zn superoxide dismutase (SOD) expression were employed for in vitro investigations. After exposure of these cells to asbestos, we demonstrated by fluorescence methodology a significantly increased generation of SOD with ferrireductive capacity. Fiber exposure also increased DMT1 protein and mRNA expression in the BEAS-2B cells. Incubation with asbestos elevated cellular iron and ferritin concentrations, and these responses were diminished in cells with an enhanced expression of SOD. Finally, fiber exposure increased supernatant concentrations of interleukin 8, but this inflammatory mediator was actually increased in cells with elevated SOD expression. We conclude that the response of respiratory epithelial cells to asbestos includes oxidant-mediated mechanisms to sequester catalytically active iron associated with the fiber.


Assuntos
Asbesto Crocidolita/toxicidade , Células Epiteliais/efeitos dos fármacos , Ferro/metabolismo , Fibras Minerais/toxicidade , Oxidantes/metabolismo , Mucosa Respiratória/efeitos dos fármacos , Proteínas de Transporte de Cátions/metabolismo , Linhagem Celular Transformada , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Ferritinas/metabolismo , Radicais Livres/metabolismo , Humanos , Interleucina-8/metabolismo , Oxirredução , Estresse Oxidativo , Mucosa Respiratória/metabolismo , Mucosa Respiratória/patologia , Superóxido Dismutase/metabolismo , Superóxidos/metabolismo
18.
J Heart Lung Transplant ; 24(11): 1821-7, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16297788

RESUMO

BACKGROUND: Oxidative stress has been proposed as a mechanism of injury underlying obliterative bronchiolitis. Catalytically reactive iron is a potential source of reactive oxygen species in transplanted tissue. Using samples acquired from surveillance bronchoalveolar lavage (BAL), we tested the postulate that there is a disruption of iron equilibrium in transplanted lung, which can worsen with time. METHODS: A control group of 5 healthy, non-smoking volunteers underwent BAL. Five bilateral lung transplant patients underwent surveillance BAL with transbronchial lung biopsies. The BAL fluid concentrations of protein, albumin, total iron, lactoferrin, ferritin, transferrin receptor and total iron binding capacity were measured. RESULTS: The mean ages in the control and transplant groups were 25.0 +/- 2.4 and 34.6 +/- 5.0 years, respectively. Patients were transplanted for cystic fibrosis (n = 3), primary ciliary dyskinesia (n = 1) and bronchiolitis obliterans (n = 1). Surveillance bronchoscopies were performed at 100.6 +/- 63.3, 175.0 +/- 87.7 and 259.2 +/- 82 days post-transplant. No significant differences were noted in BAL protein, albumin and total iron binding capacity (TIBC) levels between the 2 groups. The BAL iron, transferrin, transferrin receptor, lactoferrin and ferritin levels were significantly elevated in transplant patients relative to controls. With time after transplantation, there were increases in lavage iron, transferrin receptor, lactoferrin and ferritin concentrations. CONCLUSIONS: Abnormally high levels of iron and its homeostatic proteins were found in the lung allografts, and levels appeared to increase with time. This supports a disruption in the normal homeostasis of this metal after transplantation and a potential role for a catalyzed oxidative stress in bronchiolitis obliterans. The use of iron-depleting therapy is a possible means for preventing injury in the lung allograft.


Assuntos
Ferro/análise , Transplante de Pulmão/fisiologia , Estresse Oxidativo/fisiologia , Adolescente , Adulto , Lavagem Broncoalveolar , Fibrose Cística/cirurgia , Feminino , Ferritinas/análise , Homeostase , Humanos , Lactoferrina/análise , Masculino
19.
Am J Physiol Lung Cell Mol Physiol ; 289(1): L14-23, 2005 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-15749737

RESUMO

Ferroportin 1 (FPN1; aka MTP1, IREG1, and SLC40A1), which was originally identified as a basolateral iron transporter crucial for nutritional iron absorption in the intestine, is expressed in airway epithelia and upregulated when these cells are exposed to iron. Using immunofluorescence labeling and confocal microscopic imaging techniques, we demonstrate that in human and rodent lungs, FPN1 localizes subcellularly to the apical but not basolateral membrane of the airway epithelial cells. The role of airway epithelial cells in iron mobilization in the lung was studied in an in vitro model of the polarized airway epithelium. Normal human bronchial epithelial cells, grown on membrane supports until differentiated, were exposed to iron, and the efficiency and direction of iron transportation were studied. We found that these cells can efficiently take up iron across the apical but not basolateral surface in a concentration-dependent manner. Most of the iron taken up by the cells is then released into the medium within 8 h in the form of less reactive protein-bound complexes including ferritin and transferrin. Interestingly, iron release also occurred across the apical but not basolateral membrane. Our findings indicate that FPN1, depending on its subcellular location, could have distinct functions in iron homeostasis in different cells and tissues. Although it is responsible for exporting nutrient iron from enterocytes to the circulation in the intestine, it could play a role in iron detoxification in airway epithelial cells in the lung.


Assuntos
Proteínas de Transporte de Cátions/metabolismo , Membrana Celular/metabolismo , Células Epiteliais/metabolismo , Ferro/farmacocinética , Pulmão/fisiologia , Animais , Transporte Biológico Ativo/efeitos dos fármacos , Transporte Biológico Ativo/fisiologia , Linhagem Celular , Relação Dose-Resposta a Droga , Epitélio/metabolismo , Ferritinas/metabolismo , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/efeitos da radiação , Humanos , Inativação Metabólica/fisiologia , Mucosa Intestinal/metabolismo , Camundongos , Transferrina/metabolismo
20.
Am J Physiol Lung Cell Mol Physiol ; 289(3): L460-7, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15908475

RESUMO

Exposure to airborne particulates makes the detoxification of metals a continuous challenge for the lungs. Based on the fate of iron in airway epithelial cells, we postulated that divalent metal transporter-1 (DMT1) participates in detoxification of metal associated with air pollution particles. Homozygous Belgrade rats, which are functionally deficient in DMT1, exhibited diminished metal transport from the lower respiratory tract and greater lung injury than control littermates when exposed to oil fly ash. Preexposure of normal rats to iron in vivo increased expression of the isoform of DMT1 protein that lacked an iron-response element (-IRE), accelerated metal transport out of the lung, and decreased injury after particle exposure. In contrast, normal rats preexposed to vanadium showed less expression of the -IRE isoform of DMT1, decreased metal transport, and greater pulmonary injury after particle instillation. Respiratory epithelial cells in culture gave similar results. Also, DMT1 mRNA and protein expression for the -IRE isoform increased or decreased in these cells when exposed to iron or vanadium, respectively. These results thus demonstrate for the first time a primary role for DMT1 in lung metal transport and detoxification.


Assuntos
Proteínas de Transporte de Cátions/fisiologia , Proteínas de Ligação ao Ferro/fisiologia , Pneumopatias/induzido quimicamente , Pneumopatias/prevenção & controle , Metais , Animais , Transporte Biológico/efeitos dos fármacos , Western Blotting , Proteínas de Transporte de Cátions/deficiência , Proteínas de Transporte de Cátions/metabolismo , Linhagem Celular Transformada , Compostos Férricos/farmacologia , Imuno-Histoquímica , Ferro/farmacocinética , Proteínas de Ligação ao Ferro/metabolismo , Metais/metabolismo , Estresse Oxidativo , Isoformas de Proteínas/metabolismo , Compostos de Amônio Quaternário/farmacologia , Ratos , Ratos Endogâmicos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Vanádio/farmacocinética , Compostos de Vanádio/farmacologia
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