Long-Term Environmental Hypoxia Exposure and Haematopoietic Prolyl Hydroxylase-1 Deletion Do Not Impact Experimental Crohn’s Like Ileitis

Figures

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  Figure 1. Environmental hypoxia does not influence the general well-being of WT mice and the clinical course of gut inflammation in TNF∆ARE/+ mice. (a, left) Percentual body weight evolution of TNF∆ARE/+ mice () and their WT littermates () housed for 10 weeks in hypoxia and TNF∆ARE/+ () and WT mice () housed for 10 weeks in normoxia, all from 5 until 15 weeks of age. (a, right) Percentual body weight evolution of 8 till 15-week-old TNF∆ARE/+ mice () and their WT littermates () housed for 10 weeks in hypoxia and TNF∆ARE/+ () and WT mice () housed for 10 weeks in normoxia, all from 5 until 15 weeks of age. (b) Spleen and (c) liver weight of all mice measured at week 15. Data are represented as the mean ± standard error of the mean (SEM). N: Normoxia; H: Hypoxia. * p < 0.05.

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  Figure 2. Systemic and ileal hypoxic markers are increased in mice housed in environmental hypoxia. (a) Red blood cells, haemoglobin and haematocrit concentrations, (b) ileal vascular endothelial growth factor (Vegf) mRNA expression and (c) ileal pimonidazole intensity in WT and TNF∆ARE/+ mice housed in hypoxia and normoxia. Representative images of the pimonidazole staining in a (d) WT mouse housed in normoxia; (e) WT mouse housed in hypoxia; (f) TNF∆ARE/+ mouse housed in normoxia and (g) TNF∆ARE/+ mouse housed in hypoxia. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Data are represented as the mean ± SEM. N: Normoxia; H: Hypoxia.

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  Figure 3. Long-term environmental hypoxia exposure in WT mice induces systemic inflammation. (a) Circulatory monocytes were increased in WT mice housed in hypoxia. (b) In TNF∆ARE/+ mice housed in hypoxia, circulatory monocytes and neutrophils are increased in comparison to TNF∆ARE/+ mice housed in normoxia. Serum levels of TNF, CCL2 and IL1β in (c) WT littermates housed in normoxia and hypoxia and (d) TNF∆ARE/+ mice housed in normoxia and hypoxia. * p < 0.05, ** p < 0.01, *** p < 0.001. Data are represented as the mean ± SEM.

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  Figure 4. Long-term environmental hypoxia exposure in WT mice induces a proinflammatory ileal microenvironment. (a) Total histological inflammation score in WT mice housed in normoxia and hypoxia, comprising the subscores, (b) immune cell infiltration and (c) villi destruction. Representative images from haematoxylin-eosin (H&E) sections of the distal ileum from (d) a WT mouse housed in normoxia and (e) a WT mouse housed in hypoxia. (f) mRNA expression levels of Tnf, Ccl2 and Il1β in WT littermates housed in normoxia and hypoxia. * p < 0.05. Data are represented as the mean ± SEM. NRQ: Normalized relative quantities.

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  Figure 5. Long-term environmental hypoxia exposure to TNFΔARE/+ mice does not protect against ileitis. (a) Total histological inflammation score in TNFΔARE/+ mice housed in normoxia and hypoxia, comprising the subscores: (b) immune cell infiltration and (c) villi destruction. Representative images from H&E sections of the distal ileum from (d) a TNFΔARE/+ mouse housed in normoxia; (e) a TNFΔARE/+ mouse housed in hypoxia. (f) mRNA expression levels of Tnf, Ccl2 and Il1β in WT littermates housed in normoxia and hypoxia. Data are represented as the mean ± SEM. NRQ: Normalized relative quantities.

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  Figure 6. Haematopoietic deletion of Phd1 does not protect against ileitis in TNFΔARE/+ mice. (a) Percentual body weight evolution from week 8 till 17 of Phd1f/+Vav:cre+/− mice (), Phd1f/+ Vav:cre+/− TNF∆ARE/+ mice (), Phd1f/+Vav:cre−/− mice () and Phd1f/+Vav:cre−/− TNF∆ARE/+ mice (). (b) Total histological inflammation scores of the distal ileum from heterozygous haematopoietic Phd1 floxed mice. (c) Percentual body weight evolution from week 8 till 23 of Phd1f/fVav:cre+/− mice (), Phd1f/fVav:cre+/− TNF∆ARE/+ mice (), Phd1f/f Vav:cre−/− mice () and Phd1f/fVav:cre−/− TNF∆ARE/+ mice (). (d) Total histological inflammation scores of the distal ileum from homozygous haematopoietic Phd1 floxed mice. Representative H&E-stained ileal sections from (e) a Phd1f/fVav:cre−/− TNF∆ARE/+ and (f) a Phd1f/fVav:cre+/− TNF∆ARE/+ mouse. ** p < 0.01, *** p < 0.001, **** p < 0.0001. Data are represented as the mean ± SEM.

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  Figure 7. Haematopoietic deletion of Phd1 does not influence the proinflammatory gene expression profile in TNFΔARE/+ mice. (a) Serum levels of CCL2 and mRNA expression levels of (b) Tnf, (c) Ccl2 and (d) Il1β in the distal ileum of Phd1f/fVav:cre−/−, Phd1f/fVav:cre+/−, Phd1f/fVav:cre+/−TNFΔARE/+ and Phd1f/fVav:cre+/−TNFΔARE/+ mice. * p < 0.05; ** p < 0.01. Data are represented as the mean ± SEM. NRQ: Normalized relative quantities.