Fig 1: Effect of saliva fluid fractions, MIF, and TFF2 on development of DSS-induced colitis. (A) Coomassie-stained proteins in whole saliva fluid, (1) >100-kDa fraction, (2) >50-kDa fraction, (3) >30-kDa fraction, and (4) <30-kDa fraction. (B and C), WT (B) and Aqp5-/- mice (C) were fed with water (black) or 3% DSS (blue, green, red and dark yellow) for 6 days and allowed to recover for 9 days. The mice were gavaged with the indicated fractions of the salivary fluid on days 1, 3, and 6, as indicated. Mice were 18 to 22 weeks old, and the numbers in parenthesis indicate the number of mice in each group. The P values are relative to mice fed with DSS. The results with DSS shown in dotted lines were taken from Figure 7 and are shown to better illustrate the effect of the fractions. (D) Venn chart of the proteins in each fraction. (E) A list of the proteins in the 50-KDa fraction. MIF and TFF2 are highlighted in red. (F and G) Protocol as in (B and C), except that the mice were gavaged with 50 μL of 0.01 μg/μL MIF (green) or 0.01 μg/μL TFF2 (red). (H and I) Example images (H) and averages of intestinal length (I) measured on day 8. (J–L) Example H&E images (J) and analysis of cell proliferation (K) and goblet cell density (L). (M) Intestinal permeability expressed as fold change measured on day 8. (N–P) Example of immunofluorescence and bright field images (N) and analysis of ZO1 (O) and Occludin (P) measured on day 8. (Q and R) Intestinal (Q) and serum (R) MPO levels.
Fig 2: Scavenging saliva fluid MIF and TFF2 ameliorates effects of saliva fluid on development of DSS-induced colitis. (A and B) Standard curves (left) and the level of MIF (A) and TFF2 (B) in saliva from control (black) and DSS-treated mice (red) and saliva treated with the respective antibodies (green). (C and E) WT (C) and Aqp5-/- mice (E) were fed with water (black) or 3% DSS (blue, green and red) for 6 days and allowed to recover for 6 days. The mice were gavaged with the salivary fluid treated with anti-MIF (green) and anti-TFF2 antibodies (red) on days 1, 3, and 6, as indicated. Mice were 18 to 22 weeks old, and the numbers in parenthesis indicate the number of mice in each group. The P values are relative to mice fed with DSS. The results with DSS shown in dotted lines were taken from Figure 7 and are shown to better illustrate the effect of MIF and TFF2 depletion. (D and F) Time course of development of disease activity of the mice in (C and E), respectively. (G) Intestinal length measured on day 8. (H and I) Analysis of cell proliferation (left) and goblet cell density (right). (J and K) Intestinal (J) and serum (K) MPO levels. (L) Intestinal permeability expressed as fold change measured on day 8. (M and N) Analysis of ZO1 (M) and Occludin (N) measured on day 8.
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