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1.
Acta cir. bras ; 34(11): e201901107, Nov. 2019. graf
Artigo em Inglês | LILACS | ID: biblio-1054678

RESUMO

Abstract Purpose: To establish a hypotensive brain death pig model and observe the effects of hypotension on small bowel donors. Methods: The hypotensive brain death model was produced using the modified intracranial water sac inflation method in ten domestic crossbred pigs. Effects of hypotensive brain death on small bowel tissue morphology were evaluated through changes in intestinal tissue pathology, tight junction protein of the intestinal mucosa and plasma intestinal fatty acid-binding protein (i-FABP) levels. The pathophysiological mechanism was examined based on changes in superior mesenteric artery (SMA) blood flow and systemic hemodynamics. Results: After model establishment, SMA blood flow, and the mean arterial pressure (MAP) significantly decreased, while heart rate increased rapidly and fluctuated significantly. Small bowel tissue morphology and levels of tight junction protein of the intestinal mucosa showed that after model establishment, small bowel tissue injury was gradually aggravated over time (P<0.05). Plasma i-FABP levels significantly increased after brain death (P<0.05). Conclusions: A hypotensive brain death pig model was successfully established using an improved intracranial water sac inflation method. This method offers a possibility of describing the injury mechanisms more clearly during and after brain death.


Assuntos
Animais , Masculino , Feminino , Morte Encefálica/fisiopatologia , Modelos Animais de Doenças , Hipotensão/fisiopatologia , Intestino Delgado/patologia , Intestino Delgado/transplante , Suínos , Fatores de Tempo , Biópsia , Ensaio de Imunoadsorção Enzimática , Western Blotting , Reprodutibilidade dos Testes , Microscopia Eletrônica de Transmissão , Proteínas de Ligação a Ácido Graxo/sangue , Proteína da Zônula de Oclusão-1/análise , Hemodinâmica , Intestino Delgado/irrigação sanguínea
2.
Braz. j. med. biol. res ; 51(2): e6950, 2018. tab, graf
Artigo em Inglês | LILACS | ID: biblio-889028

RESUMO

Alveolar epithelia play an essential role in maintaining the integrity and homeostasis of lungs, in which alveolar epithelial type II cells (AECII) are a cell type with stem cell potential for epithelial injury repair and regeneration. However, mechanisms behind the physiological and pathological roles of alveolar epithelia in human lungs remain largely unknown, partially owing to the difficulty of isolation and culture of primary human AECII cells. In the present study, we aimed to characterize alveolar epithelia generated from A549 lung adenocarcinoma cells that were cultured in an air-liquid interface (ALI) state. Morphological analysis demonstrated that A549 cells could reconstitute epithelial layers in ALI cultures as evaluated by histochemistry staining and electronic microscopy. Immunofluorescent staining further revealed an expression of alveolar epithelial type I cell (AECI) markers aquaporin-5 protein (AQP-5), and AECII cell marker surfactant protein C (SPC) in subpopulations of ALI cultured cells. Importantly, molecular analysis further revealed the expression of AQP-5, SPC, thyroid transcription factor-1, zonula occludens-1 and Mucin 5B in A549 ALI cultures as determined by both immunoblotting and quantitative RT-PCR assay. These results suggest that the ALI culture of A549 cells can partially mimic the property of alveolar epithelia, which may be a feasible and alternative model for investigating roles and mechanisms of alveolar epithelia in vitro.


Assuntos
Humanos , Meios de Cultivo Condicionados , Técnicas de Cultura de Células/métodos , Células Epiteliais Alveolares/fisiologia , Células A549/fisiologia , Valores de Referência , Fatores de Tempo , Microscopia Eletrônica de Varredura , Immunoblotting , Contagem de Células , Reprodutibilidade dos Testes , Análise de Variância , Proteína C Associada a Surfactante Pulmonar/análise , Aquaporina 5/análise , Mucina-5B/análise , Reação em Cadeia da Polimerase em Tempo Real , Proteína da Zônula de Oclusão-1/análise , Fator Nuclear 1 de Tireoide/análise
3.
Experimental & Molecular Medicine ; : e167-2015.
Artigo em Inglês | WPRIM | ID: wpr-30205

RESUMO

Pulmonary dysfunction caused by ischemia-reperfusion injury is the leading cause of mortality in lung transplantation. We aimed to investigate the effects of sevoflurane pretreatment on lung permeability, tight junction protein occludin and zona occludens 1 (ZO-1) expression, and translocation of protein kinase C (PKC)-alpha after ischemia-reperfusion. A lung ischemia-reperfusion injury model was established in 96 male Wistar rats following the modified Eppinger method. The rats were divided into four groups with 24 rats in each group: a control (group C), an ischemia-reperfusion group (IR group), a sevoflurane control group (sev-C group), and a sevoflurane ischemia-reperfusion group (sev-IR group). There were three time points in each group: ischemic occlusion for 45 min, reperfusion for 60 min and reperfusion for 120 min; and there were six rats per time point. For the 120-min reperfusion group, six extra rats underwent bronchoalveolar lavage. Mean arterial pressure (MAP) and pulse oxygen saturation (SpO2) were recorded at each time point. The wet/dry weight ratio and lung permeability index (LPI) were measured. Quantitative RT-PCR and Western blot were used to measure pulmonary occludin and ZO-1, and Western blot was used to measure cytosolic and membranous PKC-alpha in the lung. Lung permeability was significantly increased after ischemia-reperfusion. Sevoflurane pretreatment promoted pulmonary expression of occludin and ZO-1 after reperfusion and inhibited the translocation of PKC-alpha. In conclusion, sevoflurane pretreatment alleviated lung permeability by upregulating occludin and ZO-1 after ischemia-reperfusion. Sevoflurane pretreatment inhibited the translocation and activation of PKC-alpha, which also contributed to the lung-protective effect of sevoflurane.


Assuntos
Animais , Masculino , Anestésicos Inalatórios/uso terapêutico , Permeabilidade Capilar/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Pneumopatias/tratamento farmacológico , Éteres Metílicos/uso terapêutico , Proteína Quinase C-alfa/metabolismo , Transporte Proteico/efeitos dos fármacos , RNA Mensageiro/genética , Ratos Wistar , Traumatismo por Reperfusão/tratamento farmacológico , Proteína da Zônula de Oclusão-1/análise
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