Quantification of the Expression of HIF-1alpha by Real-Time PCR in Rat Hepatocytes Cultures Invaded by Shigella flexneri under Normoxic and Hypoxic Conditions

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Citações na Scopus
Tipo de produção
article
Data de publicação
2015
DOI
Web of Science
Título da Revista
ISSN da Revista
Título do Volume
Editora
HANS PUBLISHERS
Autores
LIMA, Camila Barbara Cantalupo
Citação
ADVANCES IN MICROBIOLOGY, v.5, n.7, p.507-516, 2015
Projetos de Pesquisa
Unidades Organizacionais
Fascículo
Resumo
Aim: Shigella flexneri (S. flexneri) is a gram-negative enterobacterium responsible for severe intestinal end systemic infection in humans. The bacteria can reach the liver due to degeneration of the colonic epithelium. Hypoxia is present in many human diseases and can induce the expression of the transcription factor HIF-1alpha that may have a cell protective role. The influence of hypoxia and HIF-1alpha on bacterial infection, studied in this work, is unclear. Hypoxia inducible factor-1alpha (HIF-1alpha) is a transcription factor that acts as a master regulator of gene expression induced by hypoxia. Methods: We compared the ability of S. flexneri to invade rat hepatocytes in primary culture both in normoxic and hypoxic conditions. We evaluated TNF-alpha released by hepatocytes, apoptosis rate and HIF-1alpha expression by confocal microscopy as well as real time PCR technique. Results: We showed that S. flexneri invaded less hepatocytes previously submitted to 24 h hypoxia (6.5% O-2) than those cultivated in normoxia (21% O-2). S. flexneri also induced HIF-1 alpha expression in hepatocytes, TNF-alpha secretion and apoptosis. Conclusion: a) Hypoxia alone was not a stimulus to TNF-alpha secretion, but induced cell apoptosis and HIF-1 alpha expression; b) S. flexneri was able to invade rat hepatocytes and hypoxia apparently influenced significantly bacterial cell invasiveness; c) HIF-1 alpha was expressed in hypoxic conditions, and it was also stimulated by S. flexneri.
Palavras-chave
Shigella flexneri, Hepatocytes, Cell Hypoxia, Hypoxia-Inducible Factor l alpha, TNF-Alpha, Apoptosis
URI
https://observatorio.fm.usp.br/handle/OPI/29683
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Coleções
Artigos e Materiais de Revistas Científicas - HC/ICHC
Artigos e Materiais de Revistas Científicas - FM/MIP
Artigos e Materiais de Revistas Científicas - LIM/54