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. 2021 May 27;297(1):100834. doi: 10.1016/j.jbc.2021.100834

Figure 6.

Figure 6

Effects of hookworm-derived peptides in the 2,4,6-trinitrobenzenesulfonic acid (TNBS) mouse model of colitis.A, schematic overview of the experimental strategy followed. Five-week-old C57BL/6 male mice were analyzed to study the effect of different synthetic hookworm peptide doses in the intestine of mice with TNBS-induced colitis. Hookworm peptide was dosed 4 h before the administration of TNBS. Mice were monitored daily for weight loss and general well-being over 4 days before they were euthanized and their colons removed. Colons were measured in length; adhesion, edema, mucosal wall thickening, and ulceration were scored on a scale of 0 to 3, with 3 indicating the highest degree of damage. Colons were then opened up longitudinally and washed in PBS, and a 1-cm section from the distal colon was fixed in 4% paraformaldehyde. Histological micrographs of hematoxylin and eosin–stained tissue (×200) obtained from a representative mouse from each group were observed for epithelial changes (presence of goblet cells, hyperplasia, erosion, cell infiltrate and mucosal architecture). A section of colon from a representative mouse from each group was also used to assess the gene expression profile via Nanostring. B, body weight was recorded daily for the indicated groups. Data show mean ± SEM of a representative experiment out of three with n = 5. Two-way ANOVA with Dunnett’s multiple comparisons test was used to compare each group with vehicle over time. C and D, colons were removed and measured on day 4. E, macroscopic scoring of colon pathology; adhesion, edema, mucosal wall thickening, and ulceration were scored on a scale of 0 to 3, with 3 indicating the highest degree of damage. F, data show histological micrographs of hematoxylin and eosin–stained tissue (×200) obtained from a representative mouse from each group. Colons were assessed for epithelial changes (presence of goblet cells (1), hyperplasia (2), erosion, cell infiltrate (3, 4) and mucosal architecture. Data show mean ± SEM of a representative experiment with n = 5. Mann–Whitney U test was performed comparing each group with vehicle; ∗p < 0.05; ∗∗p < 0.01.