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• W2912170897 abstract "Hypoxia, a state of oxygen deficiency that occurs when oxygen concentrations fall below the normoxic atmospheric levels, has a significant effect on several cellular processes and has been shown to influence the pathogenesis of several diseases. Within the human body however, tissues are supplied with different oxygen concentrations that are below atmospheric levels but that remain biologically sufficient, thus resulting in a state termed physioxia. The gastrointestinal tract has been shown to experience a steep oxygen gradient, resulting in physioxic levels ranging between 1% and 7% in the intestinal epithelium. Therefore, when trying to establish an in vitro intestinal epithelial cell model, it is critical to properly characterize the oxygen conditions found in the cell culture.  This study aimed to measuring oxygen levels in three different systems of Caco-2 cell culture, conventional polystyrene 24-well plates, special 24-well gas permeable plates and membrane inserts, using optical sensor spots. All culture systems displayed lower O2 levels than expected when cultured under normoxic conditions over the course of six days, with conventional 24 well plates reaching as low as 3% oxygen while remaining above 13% in gas permeable plates and membrane inserts. Incubation of the different cultures under hypoxic conditions (1%) resulted in a rapid equilibration to the environmental oxygen levels in the gas permeable and conventional 24 well plates, but not in the membrane inserts that remained at 3% after 6 hours. Studies have shown that cellular hypoxia results in the accumulation of the hypoxia-inducible factor, HIF-1α, that acts a global regulator of cellular adaptation to oxygen stress. Therefore, Western blot analysis was used to determine the protein levels of HIF-1α in the different cultures, revealing a distinct stabilization of HIF-1α protein after only 1 hour of hypoxic incubation of Caco-2 cultures in conventional 24-well and gas permeable plates, and after 4 hours in membrane inserts. Therefore, we suggest that the culture system and conditions should be taken into consideration when performing oxygen-dependent experiments. Furthermore, hypoxia has been shown to occur in tissues at sites of inflammation and infection, mainly because of decreased oxygen diffusion, and increased consumption by invading pathogens and recruited immune cells. When studying bacterial infections in the gastrointestinal tract, it is important to take into account the cellular responses to low oxygen concentrations and their effect on host-pathogen interactions. This study investigates the interaction between the gastrointestinal zoonotic bacterium Yersinia enterocolitica and intestinal epithelial cells under hypoxic conditions. It was found that hypoxic pre-incubation of Caco-2 cells resulted in significantly decreased bacterial internalization, a phenotype that was abolished after functionally blocking host β1 integrins as well as upon infection with an invasin-deficient Y. enterocolitica strain. Decreased protein levels of β1 integrins in hypoxic incubated cells corroborated these results. Finally, a potential role for HIF-1α was hypothesized, since treatment with the HIF-1α agonist, dimethyloxalylglycine, also resulted in reduced Y. enterocolitica internalization and decreased β1 integrin protein levels. The study highlights the relevance of hypoxia in the context of gastrointestinal infections and discusses a role for HIF-1α as a potential therapeutic agent." @default.
• W2912170897 created "2019-02-21" @default.
• W2912170897 creator A5025608457 @default.
• W2912170897 date "2015-01-01" @default.
• W2912170897 modified "2023-09-27" @default.
• W2912170897 title "The role of hypoxia in the intestinal epithelial cell response to Yersinia enterocolitica" @default.
• W2912170897 hasPublicationYear "2015" @default.
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