RESUMO
The early inflammatory response to influenza A virus infection contributes to severe lung disease and continues to pose a serious threat to human health. The mechanisms by which inflammatory cells invade the respiratory tract remain unclear. Uncontrolled inflammation and oxidative stress cause lung damage in response to influenza A infection. We have previously shown that the fatty acid binding protein 5 (FABP5) has anti-inflammatory properties. We speculate that, as a transporter of fatty acids, FABP5 plays an important protective role against oxidative damage to lipids during infection as well. Using FABP5-/- and wild-type (WT) mice infected with influenza A virus, we showed that FABP5-/- mice had increased cell infiltration of macrophages and neutrophils compared with WT mice. FABP5-/- mice presented lower viral burden but lost as much weight as WT mice. The adaptive immune response was also increased in FABP5-/- mice as illustrated by the accumulation of T and B cells in the lung tissues and increased levels of H1N1-specific IgG antibodies. FABP5 deficiency greatly enhanced oxidative damage and lipid peroxidation following influenza A infection and presented with sustained tissue inflammation. Interestingly, FABP5 expression decreased following influenza A infection in WT lung tissues that corresponded to a decrease in the anti-inflammatory molecule PPAR-γ activity. In conclusion, our results demonstrate a previously unknown contribution of FABP5 to influenza A virus pathogenesis by controlling excessive oxidative damage and inflammation. This property could be exploited for therapeutic purposes.
Assuntos
Proteínas de Ligação a Ácido Graxo/fisiologia , Vírus da Influenza A Subtipo H1N1/patogenicidade , Influenza Humana/imunologia , Proteínas de Neoplasias/fisiologia , Estresse Oxidativo , Pneumonia/etiologia , Imunidade Adaptativa , Animais , Western Blotting , Células Cultivadas , Suscetibilidade a Doenças , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Humanos , Técnicas Imunoenzimáticas , Imunoprecipitação , Influenza Humana/complicações , Influenza Humana/virologia , Peroxidação de Lipídeos , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , PPAR gama/metabolismo , Pneumonia/metabolismo , Pneumonia/patologia , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
With a possibility for the use of chemical weapons in battlefield or in terrorist activities, effective therapies against the devastating ocular injuries, from their exposure, are needed. Oxygen plays a vital role in ocular tissue preservation and wound repair. We tested the efficacy of supersaturated oxygen emulsion (SSOE) in reducing ex vivo corneal and keratocyte injury from chloropicrin (CP). CP, currently used as a pesticide, is a chemical threat agent like the vesicating mustard agents and causes severe corneal injury. Since our previous study in human corneal epithelial cells showed the treatment potential of SSOE (55 %), we further tested its efficacy in an ex vivo CP-induced rabbit corneal injury model. Corneas were exposed to CP (700 nmol) for 2 h, washed and cultured with or without SSOE for 24 h or 96 h. At 96 h post CP exposure, SSOE treatment presented a healing tendency of the corneal epithelial layer, and abrogated the CP-induced epithelial apoptotic cell death. SSOE treatment also reduced the CP induced DNA damage (H2A.X phosphorylation) and inflammatory markers (e.g. MMP9, IL-21, MIP-1ß, TNFα). Further examination of the treatment efficacy of SSOE alone or in combination with other therapies in in vivo cornea injury models for CP and vesicants, is warranted.
Assuntos
Queimaduras Químicas/tratamento farmacológico , Córnea/efeitos dos fármacos , Queimaduras Oculares/tratamento farmacológico , Hidrocarbonetos Clorados/toxicidade , Oxigênio/farmacologia , Animais , Apoptose/efeitos dos fármacos , Queimaduras Químicas/etiologia , Queimaduras Químicas/metabolismo , Queimaduras Químicas/patologia , Córnea/metabolismo , Córnea/patologia , Citocinas/metabolismo , Dano ao DNA , Emulsões , Queimaduras Oculares/induzido quimicamente , Queimaduras Oculares/metabolismo , Queimaduras Oculares/patologia , Mediadores da Inflamação/metabolismo , Masculino , Técnicas de Cultura de Órgãos , Coelhos , Cicatrização/efeitos dos fármacosRESUMO
INTRODUCTION: Roughly 13% of all battlefield injuries include some form of ocular trauma. Ocular tissue preservation is critical for wound healing for warfighters with ocular injuries. Our team hypothesized that oxygen plays a vital role in ocular tissue preservation and wound healing and has developed a supersaturated oxygen emulsion (SOE) for the topical treatment of ocular trauma. MATERIALS AND METHODS: The partial pressure of oxygen (PO2) was measured in the SOE. Safety and efficacy studies were carried out in primary human corneal epithelial (HCE) cells, as the outermost layer is the first barrier to chemical and mechanical injury. Western blot, scratch assay, and MTT assays were conducted to determine the effect of the SOE on various molecular markers, the rate of scratch closure, and cellular viability, respectively. RESULTS: Data indicate that the SOE releases oxygen in a time-dependent manner, reaching a partial pressure within the emulsion over four times atmospheric levels. Studies in HCE cells indicate that application of the SOE does not lead to DNA damage, promote cell death, or hinder the rate of scratch closure and enhances cellular viability. Preliminary studies were carried out with chloropicrin (CP; developed as a chemical warfare agent and now a commonly used pesticide) as a chemical agent to induce ocular injury in HCE cells. CP exposures showed that SOE treatment reverses CP-induced DNA damage, apoptotic cell death, and oxidative stress markers. CONCLUSIONS: Maintaining adequate tissue oxygenation is critical for tissue preservation and wound repair, especially in avascular tissues like the cornea. Further studies examining the application of the SOE in corneal injury models are warranted.
Assuntos
Lesões da Córnea , Epitélio Corneano , Administração Tópica , Córnea , Lesões da Córnea/terapia , Emulsões , Humanos , OxigênioRESUMO
Osteoarthritis (OA) is associated with increased mechanical damage to joint cartilage. We have previously found that extracellular superoxide dismutase (ECSOD) is decreased in OA joint fluid and cartilage, suggesting oxidant damage may play a role in OA. We explored the effect of forced running as a surrogate for mechanical damage in a transgenic mouse with reduced ECSOD tissue binding. Transgenic mice heterozygous (Het) for the human ECSOD R213G polymorphism and 129-SvEv (wild-type, WT) mice were exposed to forced running on a treadmill for 45 min/day, 5 days/wk, over 8 wk. At the end of the running protocol, knee joint tissue was obtained for histology, immunohistochemistry, and protein analysis. Sedentary Het and WT mice were maintained for comparison. Whole tibias were studied for bone morphometry, finite element analysis, and mechanical testing. Forced running improved joint histology in WT mice. However, when ECSOD levels were reduced, this beneficial effect with running was lost. Het ECSOD runner mice had significantly worse histology scores compared with WT runner mice. Runner mice for both strains had increased bone strength in response to the running protocol, while Het mice showed evidence of a less robust bone structure in both runners and untrained mice. Reduced levels of ECSOD in cartilage produced joint damage when joints were stressed by forced running. The bone tissues responded to increased loading with hypertrophy, regardless of mouse strain. We conclude that ECSOD plays an important role in protecting cartilage from damage caused by mechanical loading.
Assuntos
Cartilagem Articular/fisiologia , Condicionamento Físico Animal/fisiologia , Superóxido Dismutase/metabolismo , Animais , Osso e Ossos/metabolismo , Osso e Ossos/fisiologia , Cartilagem Articular/metabolismo , Articulação do Joelho/metabolismo , Articulação do Joelho/fisiologia , Masculino , Camundongos , Camundongos Transgênicos/genética , Camundongos Transgênicos/metabolismo , Camundongos Transgênicos/fisiologia , Osteoartrite/genética , Osteoartrite/metabolismo , Osteoartrite/fisiopatologia , Polimorfismo Genético/genética , Corrida/fisiologia , Superóxido Dismutase/genéticaRESUMO
Airway obstruction in animals leads to compensation and avoidance behavior and elicits respiratory mechanosensation. The pattern of respiratory load compensation and neural activation in response to intrinsic, transient, tracheal occlusions (ITTO) via an inflatable tracheal cuff are unknown. We hypothesized that ITTO would cause increased diaphragm activity, decreased breathing frequency, and activation of neurons within the medullary and pontine respiratory centers without changing airway compliance. Obstructions were performed for 2-3 breaths followed by a minimum of 15 unobstructed breaths with an inflatable cuff sutured around the trachea in rats. The obstruction procedure was repeated for 10 min. The brains of obstructed and control animals were removed, fixed, sectioned, and stained for c-Fos. Respiratory pattern was measured from esophageal pressure (P(es)) and diaphragm electromyography (EMG(dia)). The obstructed breaths resulted in a prolonged inspiratory and expiratory time, an increase in EMG(dia) amplitude, and a more negative P(es) compared with control breaths. Neurons labeled with c-Fos were found in brain stem and suprapontine nuclei, with a significant increase in c-Fos expression for the occluded experimental group compared with the control groups in the nucleus ambiguus, nucleus of the solitary tract, lateral parabrachial nucleus, and periaqueductal gray matter. The results of this study demonstrate tracheal occlusion-elicited activation of neurons in brain stem respiratory nuclei and neural areas involved in stress responses and defensive behaviors, suggesting that these neurons mediate the load compensation breathing pattern response and may be part of the neural pathway for respiratory mechanosensation.
Assuntos
Tronco Encefálico/fisiologia , Vias Neurais/fisiologia , Neurônios/metabolismo , Centro Respiratório/fisiologia , Traqueia/fisiologia , Obstrução das Vias Respiratórias/metabolismo , Obstrução das Vias Respiratórias/fisiopatologia , Anestesia Geral , Animais , Tronco Encefálico/metabolismo , Diafragma/inervação , Diafragma/metabolismo , Diafragma/fisiologia , Eletromiografia/métodos , Masculino , Vias Neurais/metabolismo , Neurônios/fisiologia , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ratos , Ratos Sprague-Dawley , Respiração , Centro Respiratório/metabolismo , Traqueia/inervação , Traqueia/metabolismoRESUMO
Prostate cancer is the most commonly diagnosed cancer, with an estimated 240,000 new cases reported annually in the United States. Due to early detection and advances in therapies, more than 90% of patients will survive 10 years post diagnosis and treatment. Radiation is a treatment option often used to treat localized disease; however, while radiation is very effective at killing tumor cells, normal tissues are damaged as well. Potential side-effects due to prostate cancer-related radiation therapy include bowel inflammation, erectile dysfunction, urethral stricture, rectal bleeding and incontinence. Currently, radiation therapy for prostate cancer does not include the administration of therapeutic agents to reduce these side effects and protect normal tissues from radiation-induced damage. In the current study, we show that the small molecular weight antioxidant, MnTE-2-PyP, protects normal tissues from radiation-induced damage in the lower abdomen in rats. Specifically, MnTE-2-PyP protected skin, prostate, and testes from radiation-induced damage. MnTE-2-PyP also protected from erectile dysfunction, a persistent problem regardless of the type of radiation techniques used because the penile neurovascular bundles lay in the peripheral zones of the prostate, where most prostate cancers reside. Based on previous studies showing that MnTE-2-PyP, in combination with radiation, further reduces subcutaneous tumor growth, we believe that MnTE-2-PyP represents an excellent radioprotectant in combination radiotherapy for cancer in general and specifically for prostate cancer.