Effects of goat whey on cell lines.

<p>(A) Nitrite (NO) production and (B) interleukin (IL)-6 levels in Raw 264 and CMT-93 cells, respectively, in basal or LPS-stimulated conditions (100 ng/mL and 1 μg/mL, respectively). Data are expressed as the mean ± SEM. The bars with different letters are significantly different (one-way ANOVA post hoc Tukey’s test, P < 0.05).</p

Effects of goat whey on the experimental model of colitis induced by 2,4-dinitrobenzene sulfonic acid (DNBS).

<p>(A) Disease Activity Index (DAI); (B) food consumption; (C) weight/length ratio of the colon; and (D) colonic segment of the experimental groups. Data are expressed as the mean ± SEM (n = 12/group). Groups with different letters or with an asterisk (✶) differ significantly (one-way ANOVA post hoc Tukey’s test, P < 0.05).</p

Effects of goat whey on gene expression by RT-qPCR and immunofluorescence of the intestinal mucosal barrier proteins as measured.

<p>Colonic gene expression of the barrier function mediators gene expression (A) Mucin (MUC)-2, (B) MUC-3, (C) occludin, (D) zonula occludens (ZO)-1 analyzed by real-time qPCR and normalized with the housekeeping gene, Glyceraldehyde-3-phosphate dehydrohenase (GAPDH) in dinitrobenzene-sulphonic acid (DNBS) mice colitis 4 days after damage induction. Representative confocal photomicrographs of ZO-1 (E) immunoreactivity (green) in colons of the animals from each group; the sections are nuclear counterstained with DAPI (blue): (E.1) Healthy group had moderated ZO-1 labelling; (E.2) ZO-1 labelling was almost absence in DNBS control group; (E.3) ZO-1 labelling (red arrow) was strong in the treated group with goat whey; (E.4) Densitometric analysis confirmed a significant increases in ZO-1 in goat whey. Data are expressed as the means ± SEM. the groups with different letters differ significantly (one-way ANOVA post hoc Tukey’s test, P < 0.05).</p

Effects of goat whey on the colonic mucosa of colitic mice as assessed by histological examination.

<p>Sections of the colonic mucosa were stained with haematoxylin and eosin (x100): (A) Healthy, (B) DNBS control, and (C) Goat Whey. (D) Microscopic scores were assigned to the different groups according to the criteria described by Zea-Iriarte et al. (1996) [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0185382#pone.0185382.ref026" target="_blank">26</a>] and (E) Myeloperoxidase activity–MPO. Data are expressed as the means ± SEM (n = 12/group), and the groups with different letters differ significantly (one-way ANOVA post hoc Tukey’s test, P < 0.05).</p

Effects of goat whey on pro-inflammatory cytokines as measured by ELISA.

<p>Distal colon tissue samples were cultured overnight. The supernatants were assessed for cytokine levels using kits from R&D Systems (Minneapolis, MN, USA) following the manufacturer’s protocols. The cytokine levels in the supernatant were expressed as the concentration in pg/mL. (A) Interleukin (IL)-6 and (B) tumour necrosis factor (TNF)-α production in colonic tissues from mice with 2,4-dinitrobenzenesulfonic acid (DNBS)-induced colitis. Data are expressed as the mean ± SEM (n = 12). The groups with different letters are significantly different (one-way ANOVA post hoc Tukey’s test, P < 0.05).</p

Composition of goat whey (GW).

<p>Composition of goat whey (GW).</p

Immunohistochemical analysis of colonic tissue from mice with 2,4-dinitrobenzene sulfonic acid (DNBS)-induced colitis.

<p>Effects of goat whey on suppressor of cytokine signalling-1 (SOCs-1) (Panel A), inducible nitric oxide synthase (iNOS) (Panel B), NF-kappaB (NF-κB) p65 (Panel C) and p38 Mitogen Activated Protein Kinases (MAPK) (Panel D). For each antigen, three immunostained sections were examined per animal (n = 5, 3 sections per animal). 40 x magnification, scale bar = 100 μm; 1: Healthy; 2: DNBS Control; 3: Goat Whey; Arrow [⇩] = moderate marking; arrow and star [⇩★] = strong marking; star [★] = weak to moderate marking. Panel E (Immunohistochemical score)—E.1: SOCs-1; E.2: iNOS; E.3: NF-κB p65; E.4: p38 MAPK. Data are expressed as the means ± SEM; the groups with different letters differ significantly (one-way ANOVA post hoc Tukey’s test, P < 0.05).</p

Effect of goat whey on IL-17 expression in colitic mice.

<p>Representative confocal photomicrographs of IL-17 (Panel A) immunoreactivity (green) in colons of the animals from each group; the sections are nuclear counterstained with DAPI (blue): (A.1) Healthy group had absent or weak IL-17 labelling in all mucosa layers; (A.2) IL-17 labelling was strong in the DNBS control group; (A.3) weak to moderate IL-17 labelling (red arrow) was seen in the group treated with goat whey; (A.4) Densitometric analysis confirmed a significant reduction in IL-17 immunoreactivity in goat whey. Data are expressed as the means ± SEM; the groups with different letters differ significantly (one-way ANOVA post hoc Tukey’s test, P < 0.05).</p

Effects of goat whey on the gene expression of pro-inflammatory cytokines as measured by RT-qPCR.

<p>Colonic gene expression of the pro-inflammatory cytokines (A) Interleukin (IL)-1β, (B) IL-6, (C) tumour necrosis factor (TNF)-α, (D) inducible nitric oxide synthase (iNOS), (E) matrix metalloproteinase (MMP)-9, and (F) intercellular adhesion molecule (ICAM)-1 analyzed by real-time qPCR and normalized with the housekeeping gene, Glyceraldehyde-3-phosphate dehydrohenase (GAPDH) in dinitrobenzene-sulphonic acid (DNBS) mice colitis 4 days after damage induction. Data are expressed as the mean ± SEM (n = 12/group). The groups with different letters are significantly different (one-way ANOVA post hoc Tukey’s test, P < 0.05).</p