RESUMO
Acute lung injury (ALI) is generally caused by severe respiratory infection and characterized by overexuberant inflammatory responses and inefficient pathogens-containing, the two major processes wherein alveolar macrophages (AMs) play a central role. Dysfunctional mitochondria have been linked with distorted macrophages and hence lung disorders, but few treatments are currently available to correct these defects. Plant-derive nanovesicles have gained significant attention because of their therapeutic potential, but the targeting cells and the underlying mechanism remain elusive. We herein prepared the nanovesicles from Artemisia annua, a well-known medicinal plant with multiple attributes involving anti-inflammatory, anti-infection, and metabolism-regulating properties. By applying three mice models of acute lung injury caused by bacterial endotoxin, influenza A virus (IAV) and SARS-CoV-2 pseudovirus respectively, we showed that Artemisia-derived nanovesicles (ADNVs) substantially alleviated lung immunopathology and raised the survival rate of challenged mice. Macrophage depletion and adoptive transfer studies confirmed the requirement of AMs for ADNVs effects. We identified that gamma-aminobutyric acid (GABA) enclosed in the vesicles is a major molecular effector mediating the regulatory roles of ADNVs. Specifically, GABA acts on macrophages through GABA receptors, promoting mitochondrial gene programming and bioenergy generation, reducing oxidative stress and inflammatory signals, thereby enhancing the adaptability of AMs to inflammation resolution. Collectively, this study identifies a promising nanotherapeutics for alleviating lung pathology, and elucidates a mechanism whereby the canonical neurotransmitter modifies AMs and mitochondria to resume tissue homeostasis, which may have broader implications for treating critical pulmonary diseases such as COVID-19.
Assuntos
Lesão Pulmonar Aguda , Plantas Medicinais , Pneumonia Viral , Pneumonia , Camundongos , Animais , Macrófagos Alveolares/metabolismo , Pulmão/metabolismo , Pneumonia Viral/tratamento farmacológico , Lesão Pulmonar Aguda/patologia , Mitocôndrias/patologia , Ácido gama-Aminobutírico/metabolismo , Pneumonia/metabolismoRESUMO
Nitrogen mustard (NM) is a toxic vesicant that causes acute injury to the respiratory tract. This is accompanied by an accumulation of activated macrophages in the lung and oxidative stress which have been implicated in tissue injury. In these studies, we analyzed the effects of N-acetylcysteine (NAC), an inhibitor of oxidative stress and inflammation on NM-induced lung injury, macrophage activation and bioenergetics. Treatment of rats with NAC (150 mg/kg, i.p., daily) beginning 30 min after administration of NM (0.125 mg/kg, i.t.) reduced histopathologic alterations in the lung including alveolar interstitial thickening, blood vessel hemorrhage, fibrin deposition, alveolar inflammation, and bronchiolization of alveolar walls within 3 d of exposure; damage to the alveolar-epithelial barrier, measured by bronchoalveolar lavage fluid protein and cells, was also reduced by NAC, along with oxidative stress as measured by heme oxygenase (HO)-1 and Ym-1 expression in the lung. Treatment of rats with NAC attenuated the accumulation of macrophages in the lung expressing proinflammatory genes including Ptgs2, Nos2, Il-6 and Il-12; macrophages expressing inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and tumor necrosis factor (TNF)α protein were also reduced in histologic sections. Conversely, NAC had no effect on macrophages expressing the anti-inflammatory proteins arginase-1 or mannose receptor, or on NM-induced increases in matrix metalloproteinase (MMP)-9 or proliferating cell nuclear antigen (PCNA), markers of tissue repair. Following NM exposure, lung macrophage basal and maximal glycolytic activity increased, while basal respiration decreased indicating greater reliance on glycolysis to generate ATP. NAC increased both glycolysis and oxidative phosphorylation. Additionally, in macrophages from both control and NM treated animals, NAC treatment resulted in increased S-nitrosylation of ATP synthase, protecting the enzyme from oxidative damage. Taken together, these data suggest that alterations in NM-induced macrophage activation and bioenergetics contribute to the efficacy of NAC in mitigating lung injury.
Assuntos
Acetilcisteína , Metabolismo Energético , Lesão Pulmonar , Mecloretamina , Estresse Oxidativo , Animais , Estresse Oxidativo/efeitos dos fármacos , Acetilcisteína/farmacologia , Mecloretamina/toxicidade , Masculino , Metabolismo Energético/efeitos dos fármacos , Ratos , Lesão Pulmonar/induzido quimicamente , Lesão Pulmonar/metabolismo , Lesão Pulmonar/patologia , Ratos Sprague-Dawley , Pulmão/efeitos dos fármacos , Pulmão/metabolismo , Pulmão/patologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/metabolismo , Lesão Pulmonar Aguda/patologia , Macrófagos Alveolares/efeitos dos fármacos , Macrófagos Alveolares/metabolismo , Substâncias para a Guerra Química/toxicidadeRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Bletilla striata polysaccharides (BSP) extracted from the B. striata tuber, have been demonstrated to possess anti-inflammatory properties. However, their potential protective effect against ARDS and their role in regulating cell pyroptosis remained unexplored. AIM OF THE STUDY: The aim of this study was to investigate the therapeutic effect of BSP in the alleviation of lipopolysaccharide (LPS)-induced ARDS, and to explore its mechanism of action. METHODS: The effect of BSP was assessed by LPS injection into the intraperitoneal cavity in vivo; pathological changes of ARDS mice were gauged by immunohistochemical, hematoxylin and eosin staining, and immunofluorescence assays. MH-S cells were used to model the pyroptosis in vitro. Finally, the pyroptosis of alveolar macrophage was detected by western blots, qPCR, and flow cytometry for NLRP3/caspase1/GSDMD and HMGB1/TLR4 pathway-associated proteins and mRNA. RESULTS: BSP could significantly increase the weight and survival rate of mice with ARDS, alleviate the cytokine storm in the lungs, and reduce lung damage in vivo. BSP inhibited the inflammation caused by LPS/Nigericin significantly in vitro. Compared with the control group, there was a remarkable surge in the incidence of pyroptosis observed in ARDS lung tissue and alveolar macrophages, whereas BSP significantly diminished the pyroptosis ratio. Besides, BSP reduced NLRP3/caspase1/GSDMD and HMGB1/TLR4 levels in ARDS lung tissue and MH-S cells. CONCLUSIONS: These findings proved that BSP could improve LPS-induced ARDS via inhibiting pyroptosis, and this effect was mediated by NLRP3/caspase1/GSDMD and HMGB1/TLR4, suggesting a therapeutic potential of BSP as an anti-inflammatory agent for ARDS treatment.
Assuntos
Proteína HMGB1 , Síndrome do Desconforto Respiratório , Animais , Camundongos , Macrófagos Alveolares , Lipopolissacarídeos/toxicidade , Proteína 3 que Contém Domínio de Pirina da Família NLR , Piroptose , Receptor 4 Toll-Like , Polissacarídeos/farmacologia , Polissacarídeos/uso terapêutico , PulmãoRESUMO
SCOPE: Particulate matter (PM) contains toxic organic matter and heavy metals that enter the entire body through blood flow and may cause mortality. Ganoderma formosanum mycelium, a valuable traditional Chinese medicine that has been used since ancient times, contains various active ingredients that can effectively impede inflammatory responses on murine alveolar macrophages induced by PM particles. METHODS AND RESULTS: An experimental study assessing the effect of G. formosanum mycelium extract's water fraction (WA) on PM-exposed murine alveolar macrophages using ROS measurement shows that WA reduces intracellular ROS by 12% and increases cell viability by 16% when induced by PM particles. According to RNA-Sequencing, western blotting, and real-time qPCR are conducted to analyze the metabolic pathway. The WA reduces the protein ratio in p-NF-κB/NF-κB by 18% and decreases the expression of inflammatory genes, including IL-1ß by 38%, IL-6 by 29%, and TNF-α by 19%. Finally, the identification of seven types of anti-inflammatory compounds in the WA fraction is achieved through UHPLC-ESI-Orbitrap-Elite-MS/MS analysis. These compounds include anti-inflammatory compounds, namely thiamine, adenosine 5'-monophosphate, pipecolic acid, L-pyroglutamic acid, acetyl-L-carnitine, D-mannitol, and L-malic acid. CONCLUSIONS: The study suggests that the WA has the potential to alleviate the PM -induced damage in alveolar macrophages, demonstrating its anti-inflammatory properties.
Assuntos
Ganoderma , Macrófagos Alveolares , NF-kappa B , Camundongos , Animais , Macrófagos Alveolares/química , Macrófagos Alveolares/metabolismo , NF-kappa B/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Espectrometria de Massas em Tandem , Material Particulado/toxicidade , Material Particulado/análise , Anti-Inflamatórios/farmacologia , Pulmão/química , Pulmão/metabolismoRESUMO
BACKGROUND: Alveolar macrophages are one of the momentous regulators in pulmonary inflammatory responses, which can secrete extracellular vesicles (EVs) packing miRNAs. Ferroptosis, an iron-dependent cell death, is associated with cigarette smoke-induced lung injury, and EVs have been reported to regulate ferroptosis by transporting intracellular iron. However, the regulatory mechanism of alveolar macrophage-derived EVs has not been clearly illuminated in smoking-related pulmonary ferroptosis. Despite the known anti-ferroptosis effects of naringenin in lung injury, whether naringenin controls EVs-mediated ferroptosis has not yet been explored. PURPOSE: We explore the effects of EVs from cigarette smoke-stimulated alveolar macrophages in lung epithelial ferroptosis, and elucidate the EV miRNA-mediated pharmacological mechanism of naringenin. STUDY DESIGN AND METHODS: Differential and ultracentrifugation were conducted to extract EVs from different alveolar macrophages treatment groups in vitro. Both intratracheal instilled mice and treated epithelial cells were used to investigate the roles of EVs from alveolar macrophages involved in ferroptosis. Small RNA sequencing analysis was performed to distinguish altered miRNAs in EVs. The ferroptotic effects of EV miRNAs were examined by applying dual-Luciferase reporter assay and miRNA inhibitor transfection experiment. RESULTS: Here, we firstly reported that EVs from cigarette smoke extract-induced alveolar macrophages (CSE-EVs) provoked pulmonary epithelial ferroptosis. The ferroptosis inhibitor ferrostatin-1 treatment reversed these changes in vitro. Moreover, EVs from naringenin and CSE co-treated alveolar macrophages (CSE+Naringenin-EVs) markedly attenuated the lung epithelial ferroptosis compared with CSE-EVs. Notably, we identified miR-23a-3p as the most dramatically changed miRNA among Normal-EVs, CSE-EVs, and CSE+Naringenin-EVs. Further experimental investigation showed that ACSL4, a pro-ferroptotic gene leading to lipid peroxidation, was negatively regulated by miR-23a-3p. The inhibition of miR-23a-3p diminished the efficacy of CSE+Naringenin-EVs. CONCLUSION: Our findings firstly provided evidence that naringenin elevated the EV miR-23a-3p level from CSE-induced alveolar macrophages, thereby inhibiting the mouse lung epithelial ferroptosis via targeting ACSL4, and further complemented the mechanism of cigarette-induced lung injury and the protection of naringenin in a paracrine manner. The administration of miR-23a-3p-enriched EVs has the potential to ameliorate pulmonary ferroptosis.
Assuntos
Fumar Cigarros , Vesículas Extracelulares , Ferroptose , Flavanonas , Lesão Pulmonar , MicroRNAs , Camundongos , Animais , Macrófagos Alveolares/metabolismo , Fumar Cigarros/efeitos adversos , Pulmão/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Vesículas Extracelulares/metabolismo , Ferro/metabolismoRESUMO
Introdução: Este estudo foi realizado com o intuito de avaliar efeitos da acupuntura sobre os pacientes com asma leve e moderada persistentes com o uso de beta-2 agonista ou corticoide inalatório. Métodos e casuística: Trata-se de um estudo prospectivo, duplo-cego, randomizado e cruzado com dois braços. Os 74 pacientes com diagnóstico de asma leve/moderada, de acordo com a classificação de GINA 2002/2003, foram divididos em dois grupos, sendo 31 do Grupo I, e 43 do Grupo II inicialmente. Foram realizadas consultas médicas e exames que incluíram espirometria, citologia de escarro induzido, NO expirado, preenchimento de escala de sintoma, questionários de qualidade de vida de asma e de SF 36, e realização de peak-flow, dependendo da Fase do protocolo. A Fase I constituiu-se dos exames pré-intervenção. Na Fase II, foram realizadas 10 sessões de Acupuntura Real no Grupo I e 10 sessões de Acupuntura Sham no Grupo II, na Fase III, houve 4 semana de washout, na Fase IV, houve a troca de técnicas de acupuntura, sendo uma sessão por semana e, na Fase V, realização dos exames. Resultados: Não há diferença nos critérios de avaliação no pré-tratamento entre dois grupos, com exceção de maior celularidade inflamatória no Grupo II. No entanto, houve uma redução significativa de eosinófilos (p = 0,035) e neutrófilos (p = 0,047), e aumento de macrófagos (p = 0,001), melhora da medida de volume do peak-flow (p = 0,01) na fase IV do Grupo II. No Grupo I, na avaliação de escala de sintomas diária, havia menor uso de medicação de resgate (p = 0,043) na Fase II, e, depois de receber a Acupuntura Sham na Fase IV, havia menos tosse (p = 0,007), menos chiado (p = 0,037), menos dispneia (p < 0,001) e menor uso de medicação de resgate (p < 0,001). No Grupo II, após receber o tratamento com a Acupuntura Sham na Fase II, houve diminuição de tosse (p = 0,037), de chiado (p = 0,013) e de dispneia (p = 0,014), e, na...
Introduction: This survey has been conducted in order to evaluate the effects of acupuncture in patients with persistent mild and moderate asthma (according to GINA criteria 2003), using beta agonist and/or inhaled glucocorticoid. Methods and patients: This is a prospective, double blinded, randomized and cross-over study with two branches: 74 patients diagnosed with mild and moderate asthma were divided into two groups: Group I with 31, initiating with real acupuncture and Group II, starting with sham acupuncture. Medical interview and laboratory tests including spirometry, induced sputum citology, exhaled NO measurement, quality of life questionnaire (SF-36 and QQL), besides, daily symptom scores and measurement of peak-flow were performed, in the beginning of the study, and in the end of each phase of treatment. Phase I: laboratory tests and other qualitative measurements. There were 10 real acupuncture weekly sessions to Group I and 10 sham acupuncture sessions to Group II in Phase II. On the other hand, in the Phase IV, there was an exchange between Group I and Group II, which was receiving real acupuncture started to receive sham, and vice-versa, the number of sessions remained the same (10 weekly sessions). Phase III, during the interval between Phase II and Phase IV, there was an interval of 4 weeks of washout. Phase V: laboratory tests and other qualitative measurements. Results: There was no difference beween both the groups in all criteria of evaluation pré treatment, with only na exception: in the Group II there was large inflammatory cell counts. However, there was a significant reduction in eosinophils (p = 0.035) and neutrophils (p = 0.047), and increase of macrophages (p = 0.001), improved peak-flow measurement in the morning (p = 0.01) in Group II (started with sham) in Phase IV. In Daily Symptons Score, there was a significant reduction in use of rescue medication (p = 0.043) in Group I (real acupuncture) in Phase II and after received...
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Acupuntura , Terapia por Acupuntura , Asma , Asma/imunologia , Dispneia/prevenção & controle , Eosinófilos , Macrófagos Alveolares , Medicina Tradicional Chinesa/psicologia , Neutrófilos , Perfil de Impacto da Doença , Ensaio Clínico Controlado , Sinais e Sintomas Respiratórios , Sintomas Afetivos/imunologiaRESUMO
<p><b>OBJECTIVE</b>To investigate the anti-inflammatory and immunoregulatory effects of Yupingfeng (, YPF) Powder and its components in rats.</p><p><b>METHODS</b>A rat chronic bronchitis (CB) model was developed using lipopolysaccharide (LPS) combined with bacillus Calmette Guerin (BCG). YPF, simple recipe Astragalus membranaceus (Fisch.) Bge (AM) and Astragalus membranaceus (Fisch.) Bge plus rhizome of Atractylodes macrocephala Koidz (AM+RA) decoction were administered (intragastric administration, once a day for 21 days) to rats, to prevent and treat CB. Immunoregulatory and anti-inflammatory effects of YPF, AM and AM+RA were tested by serum pharmacology in vitro on splenic lymphocytes of normal rats and alveolar macrophages of CB rats.</p><p><b>RESULTS</b>Inflammation in the pulmonary tissue and the bronchus of CB rats was significantly reduced in the YPF-treatment groups, AM and AM+RA groups demonstrating the efficacy of YPF. Serum samples collected at different times from rats after administration of YPF, AM and AM+RA demonstrated increased proliferation of splenic lymphocytes with area under the effect curve (AUE) of 552.6%, 336.3% and 452.0%, respectively. Treatment of alveolar macrophages with serum samples in YPF, AM or AM+RA group inhibited interleukin-8 (IL-8) in the cell culture media, and the effect was much better in the YPF group compared with AM or AM+RA group, with a higher maximal effect (Emax, P<0.05) and larger AUE (P <0.01 and P<0.05). Moreover, serum from rats treated with AM or AM+RA had similar efficacy, while the efficiency was lower than that treated with YPF.</p><p><b>CONCLUSION</b>YPF demonstrated anti-inflammatory and immunoregulatory effects in a rat model of CB, and timedependent relationships were demonstrated in vitro.</p>
Assuntos
Animais , Ratos , Anti-Inflamatórios , Farmacologia , Usos Terapêuticos , Peso Corporal , Bronquite Crônica , Tratamento Farmacológico , Patologia , Proliferação de Células , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas , Farmacologia , Usos Terapêuticos , Fatores Imunológicos , Farmacologia , Usos Terapêuticos , Interleucina-8 , Metabolismo , Pulmão , Patologia , Linfócitos , Macrófagos Alveolares , Metabolismo , Pós , Ratos Sprague-Dawley , Baço , Patologia , Fatores de TempoRESUMO
<p><b>OBJECTIVE</b>To study the effects of huai qi huang, a traditional Chinese medicine, on cytokines Th1, Th2 and Th17 levels and alveolar macrophage phagocytosis in asthmatic rats sensitized by ovalbumin (OVA).</p><p><b>METHODS</b>Forty male Sprague-Dawley rats were randomly divided into five groups: normal control, untreated asthma, budesonide-treated, huai qi huang-treated and budesonide+huai qi huang-treated asthma (n=8 each). Asthma was induced by OVA sensitization and challenge. The levels of IL-4, IFN-γ and IL-17 in plasma and bronchoalveolar lavage fluid (BALF) were measured using ELISA. The phagocytosis of alveolar macrophages which were isolated and purified from BALF was evaluated by the colorimetric assay.</p><p><b>RESULTS</b>The levels of IL-4 and IL-17 increased, in contrast, the IFN-γ level decreased in plasma and BALF in the untreated asthma group compared with those in the normal control group. The IFN-γ level in the huai qi huang-treated asthma group was higher than that in the untreated asthma group. The IFN-γ level increased and the IL-17 level decreased more significantly in the budesonide+huai qi huang-treated asthma group when compared with the budesonide and huai qi huang alone treatment groups. The phagocytosis of alveolar macrophages in the untreated asthma group was lower than that in the normal control group. Huai qi huang alone or combined with budesonide increased the phagocytosis of alveolar macrophages compared with the normal control, untreated asthma and budesonid-treated asthma groups. The levels of IFN-γ in plasma and BALF were positively correlated with the phagocytosis of alveolar macrophages.</p><p><b>CONCLUSIONS</b>The levels of IL-4 and IL-17 increase and the IFN-γ level decreases in plasma and BALF, and the phagocytosis of alveolar macrophages decreases in asthmatic rats. Huai qi huang treatment may increase the IFN-γ expression in plasma and BALF and the phagocytosis of alveolar macrophages in asthmatic rats. There is a synergistic effect between huai qi huang and glucocorticoids.</p>
Assuntos
Animais , Masculino , Ratos , Asma , Tratamento Farmacológico , Alergia e Imunologia , Citocinas , Macrófagos Alveolares , Alergia e Imunologia , Medicina Tradicional Chinesa , Fagocitose , Ratos Sprague-Dawley , Linfócitos T Auxiliares-Indutores , Alergia e Imunologia , Células Th1 , Alergia e Imunologia , Células Th17 , Alergia e Imunologia , Células Th2 , Alergia e ImunologiaRESUMO
<p><b>OBJECTIVE</b>To observe the effect of Ginkgo biloba extract (GbE) against the bleomycin induced pulmonary fibrosis and to investigate its mechanisms.</p><p><b>METHOD</b>SD rats were randomly divided into 3 groups: Control group, BLM + NS group and BLM + GbE group (n = 6 in each time point of each group). Rats in each group were sacrificed on the 14th and 30th day after endotracheal injection of bleomycin A5. Lung injury through HE stain and pulmonary fibrosis through Masson stain were observed by light microscope. The content of collagen protein in lung tissue and malondialdehyde (MDA) in plasma were assayed by biochemical methods. The expressions of TGF-beta1 in tissue and bronchoalveolar lavage fluid (BALF) were detected by immunohistochemistry and immunocytochemistry respectively.</p><p><b>RESULT</b>Compared with control group, every datum in each time point in BLM + NS group showed significant changes which indicated the success of the model. Compared with BLM + NS (14 d) group, MDA in serum and TGF-beta1 in alveolar macrophage were significantly reduced in BLM + GbE (14 d) group. The data in BLM + GbE (30 d) group were compared with those in BLM + NS (30 d) group as follows. The lung injury and fibrosis were significantly ameliorated, the content of collagen in lung tissue and MDA in plasma were significantly reduced, the expression of TGF-beta1 in lung tissue was significantly reduced, however the expression of TGF-beta1 in BALF cells was not significantly changed.</p><p><b>CONCLUSION</b>GbE inhibited bleomycin induced lung injury and fibrosis in rats. The possible mechanisms were that GbE could inhibit the expression of TGF-beta1 in alveolar macrophage in early stage of fibrosis (14 d) and in noninflammatory cells in proliferative stage (30 d), and GbE could also attenuate the oxidative stress induced by bleomycin.</p>
Assuntos
Animais , Humanos , Masculino , Ratos , Bleomicina , Líquido da Lavagem Broncoalveolar , Química , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas , Ginkgo biloba , Química , Macrófagos Alveolares , Metabolismo , Malondialdeído , Metabolismo , Fibrose Pulmonar , Tratamento Farmacológico , Genética , Metabolismo , Distribuição Aleatória , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta1 , Genética , MetabolismoRESUMO
OBJETIVO: Avaliar a liberação de ânion superóxido por macrófagos alveolares em ratos submetidos ou não ao estresse agudo, ao exercício físico de natação e à suplementação com glutamina. MÉTODOS: Quarenta e dois ratos machos da linhagem Wistar com idade em torno de 62 (desvio-padrão=3) dias de idade foram divididos em grupos controle, treino, estresse e glutamina. Após a intervenção, macrófagos alveolares foram coletados e estimulados com acetato de formol miristato para a avaliação da liberação de ânion superóxido. RESULTADOS: Em comparação à primeira hora (controle=26,2, desvio-padrão=4,2; treino=28,7, desvio-padrão=5,1; estresse=20,3, desvio-padrão=4,4; glutamina=26,2, desvio-padrão=4,2), houve aumento (p<0,001) da liberação de superóxido em todos os grupos experimentais na segunda hora (controle=38,4, desvio-padrão=4,9; treino=40,7, desvio-padrão=6,1; estresse=30,2, desvio-padrão=5,6; glutamina=39,2, desvio-padrão=5,2) de observação. O treinamento e a suplementação com glutamina não provocaram diferenças na liberação de superóxido em macrófagos alveolares quando comparados ao grupo controle. Apenas nos ratos submetidos a estresse houve redução da liberação de superóxido tanto na primeira (20,3, desvio-padrão=4,4; p<0,05) quanto na segunda hora (30,2, desvio-padrão=5,6; p<0,05) de observação. CONCLUSÃO: Os achados sugerem que o estresse pode ser um dos fatores implicados na imunossupressão, uma vez que a redução da produção de ânion superóxido por macrófagos pode levar à diminuição de sua capacidade microbicida. No entanto, o protocolo de treinamento físico de natação usado e a suplementação com glutamina, na quantidade e na forma administrada, não alteraram a liberação de superóxido por macrófagos alveolares.
OBJECTIVE: To assess the release of superoxide anion from alveolar macrophages of rats submitted or not to acute restraint stress, forced swimming and glutamine supplementation. METHODS: Forty-two male Wistar rats aging roughly 62 days (standard deviation=3) were randomly divided into four groups: control, training, stress and glutamine. After the intervention, alveolar macrophages were collected and stimulated with phorbol myristate acetate to assess the release of superoxide anion. RESULTS: When compared with the first hour (control=26.2, standard deviation=4.2; training=28.7, standard deviation=5.1; stress=20.3 , standard deviation=4.4; glutamine=26.2, standard deviation=4.2), the release of superoxide increased (p<0.001) in all experimental groups in the second hour (control=38.4, standard deviation=4.9; training=40.7, standard deviation=6.1; stress=30.2, standard deviation=5.6; glutamine=39.2, standard deviation=5.2) of observation. Training and glutamine supplementation did not induce differences in the release of superoxide from alveolar macrophages when compared with the control group. Only the rats submitted to stress showed a reduction in the release of superoxide in both the first (20.3, standard deviation=4.4; p<0.05) and second hours (30.2, standard deviation=5.6; p<0.05) of observation. CONCLUSION: The results suggest that stress can be one of the factors associated with immunosuppression since reduced release of superoxide anion from macrophages can lead to reduced microbicidal capacity. On the other hand, the swimming protocol we used and the amount and route of glutamine supplementation did not change the release of superoxide from alveolar macrophages.
Assuntos
Animais , Masculino , Ratos , Estresse Mecânico , Glutamina/administração & dosagem , Glutamina/metabolismo , Macrófagos Alveolares , Natação , Superóxidos , Ratos WistarRESUMO
Macrófagos alveolares de rata se obtuvieron mediante lavado pulmonar, y se usaron como modelo para estudiar el daño en la membrana causado por exposición a ozono (O3, o,94 ppm durante 60 min), usando como índice la liberación de Cr, cargado previamente en las células, a preincubación de los macrófagos en presencia de vitamina C (ascorbato de sodio) o de vitamina E (DL tocoferol) disminuyó significativamente la liberación de CR en respuesta a O3; el efecto protector fue más pronunciado al aumentar la dosis de vitamina E. Se presenta una explicación del efecto protector de las vitaminas E y C, y se propone una hipótesis que relacionaría nuestros hallazgos con otros recientes que demuestran que la ingesta de suplementos antioxidantes contribuye a mantenimiento de la funcion pulmonar en sujetos voluntarios normales y asmáticos expuestos a ozono.
The damaging effects of a 60 minute ozone exposure (0,594 ppm) on the cell membrane of rat alveolar macrophages was assessed by measuring specific release of51Cr label from the cells. Preincubation of the macrophages in the presence of vitamin C (sodium ascorbate) or vitamin E (DL tocoferol) prior to ozone exposure significantly diminished 51Cr release. The protective effect of vitamin E was dose dependent. A proposal accounting for the protective effect of vitamins E and C on the cell membrane is presented, and our findings are discussed in relation to recent reports showing that antioxidant supplementation contributes to preserve pulmonary function in ozone-exposed normal and asthmatic volunteers.
Assuntos
Animais , Ratos , Ácido Ascórbico/uso terapêutico , Macrófagos Alveolares , Ozônio , Vitamina E , Costa RicaRESUMO
OBJECTIVES: This study was conducted to evaluate the cytotoxicity of gallium arsenide(GaAs), indium phosphide(InP) and indium arsenide(InAs) all of which are used as the semiconductor eletments in semiconductor industry. METHODS: Cytotoxicity in the alveolar macrophage was evaluated by the measurement of in vitro magnetometry, LDH release assay and histological examination. RESULTS: The relaxation curves by the in vitro magnetometry showed that GaAs has the cytotoxicity for the alveolar macrophage which is more significant in the higher dosages, while this cytotoxicity is not appeared in the groups added with InP or InAs or PBS. In the decay constant for two minutes after magnetization, GaAs-added groups showed a significant decrease with increasing doses, but both InP- and InAs-added groups did not show any significance. The LDH release assay showed a dose-dependent increasing tendency in the GaAs-, InP- and InAs-added groups. In terms of cellular morphological changes, GaAs-added groups revealed such severe cellular damages as prominent destructions in cell membranes and their morphological changes of nucleus, while InP- and InAs-added groups remained intact in intracellular structures, except for cytoplasmic degenerations. CONCLUSIONS: It is suggested that GaAs is more influential to cytotoxicity of alveolar macrophages than InP and InAs.
Assuntos
Arsênio , Arsenicais , Membrana Celular , Citoplasma , Gálio , Índio , Macrófagos , Macrófagos Alveolares , Magnetometria , Relaxamento , SemicondutoresRESUMO
Inducible nitric oxide synthase (iNOS) expression of several organs on the lipopolysaccharides (LPS)-injected rats and on excisional wound was observed by immunohistochemical methods to investigate iNOS-positive cells during inflammation. iNOS expression was induced in response to LPS in the brain and these reactions were observed in the choroidal epithelium, ependymal cells and a few of nerve cells and fiber. A more intensive reaction of nerve cell and fiber was mainly observed in the corpus callosum and hypothalamus. Induction of iNOS of the lung was observed in alveolar macrophage, smooth muscle, pneumocytes and inflammatory cells infilterated in the alveolar septum. iNOS expression of the liver was detected in Kupffer cells, hepatocytes, bile duct and inflammatory cells of spotty necrosis. The cardiac muscle and endothelial cell of the heart showed positive iNOS expression. In the excisional wound, inflammatory cells including macrophages, neutrophil and fibrobast showed iNOS expression and mainly detected necrobiotic layer. Collectively, iNOS expression was induced in the several cell types during inflammatory process. So for better understanding the function of iNOS, more research should be done in relation to each cell type of organ.
Assuntos
Animais , Ratos , Ductos Biliares , Encéfalo , Corioide , Corpo Caloso , Células Endoteliais , Epitélio , Coração , Hepatócitos , Hipotálamo , Inflamação , Células de Kupffer , Lipopolissacarídeos , Fígado , Pulmão , Macrófagos , Macrófagos Alveolares , Músculo Liso , Miocárdio , Necrose , Neurônios , Neutrófilos , Óxido Nítrico Sintase Tipo II , Células Epiteliais Alveolares , Choque , Cicatrização , Ferimentos e LesõesRESUMO
BACKGROUND: In the pathogenesis of acute lung injury induced by lipopolysaccharide(LPS), oxygen radiclls are known to be involved in one part. Superoxide dismutase(SOD) protects oxygen radical-induced tissue damage by dismutating superoxide to hydrogen peroxide. In eukaryotic cells, two forms of SOD exist intracellularly as a cytosolic, dimeric copper/zinc-containing SOD (CuZnSOD) and a mitochondrial, tetrameric manganese-containing SOD(MnSOD). But there has been little information about SOD gene expression and its regulation in pulmonary alveolar macrophages(PAMs). The objective of this study is to evaluate the SOD gene expression induced by LPS and its regulation in PAMs of rat. METHOD: In Sprague-Dawley rats, PAMs obtained by broncholaveolar lavage were purified by adherence to plastic plate. To study the effect of LPS on the SOD gene expression of PAMs, they were stimulated with different doses of LPS(0.01microg/ml ~ 10microg/ml) and for different intervals(0, 2, 4, 8, 24hrs). Also for evaluating the level of SOD gene regulation actinomycin D(AD) or cyclo- heximide(CHX) were added respectively. To assess whether LPS altered SOD mRNA stability, the rate of mRNA decay was determined in control group and LPS-treated group. Total cellular RNA extraction by guanidinium thiocyanate/phenol/chloroform method and Northern blot analysis by using a 32P-labelled rat MnSOD and CuZnSOD cDNAs were performed. RESULTS: The expression of mRNA in MnSOD increased dose-dependently, but not in CuZnSOD. MnSOD mRNA expression peaked at 8 hours after LPS treatment. Upregulation of MnSOD mRNA expression induced by LPS was suppressed by adding AD or CHX respectively. MnSOD mRNA stability was not altered by LPS. CONCLUSION: These findings show that PAMs of rat could be an important source of SOD in response to LPS, and suggest that their MnSOD mRNA expression may be regulated transcriptionally and require de novo protein synthesis without affecting mRNA stability.