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. 2022 Sep 16;14(18):3833.
doi: 10.3390/nu14183833.

Barley Leaf Ameliorates Citrobacter rodentium-Induced Colitis through Preventive Effects

Yu Feng  1 , Daotong Li  1 , Chen Ma  1 , Meiling Tian  1 , Xiaosong Hu  1 , Fang Chen  1
Affiliations

Barley Leaf Ameliorates Citrobacter rodentium-Induced Colitis through Preventive Effects

Yu Feng et al. Nutrients. .

Abstract

The incidence and prevalence of inflammatory bowel disease (IBD) have been increasing globally and progressively in recent decades. Barley leaf (BL) is a nutritional supplement that is shown to have health-promoting effects on intestinal homeostasis. Our previous study demonstrated that BL could significantly attenuate Citrobacter rodentium (CR)-induced colitis, but whether it exerts a prophylactic or therapeutic effect remains elusive. In this study, we supplemented BL before or during CR infestation to investigate which way BL acts. The results showed that BL supplementation prior to infection significantly reduced the disease activity index (DAI) score, weight loss, colon shortening, colonic wall swelling, and transmissible murine colonic hyperplasia. It significantly reduced the amount of CR in the feces and also markedly inhibited the extraintestinal transmission of CR. Meanwhile, it significantly reduced the levels and expression of tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFNγ), and interleukin-1β (IL1β). In addition, pretreatment with BL improved CR-induced gut microbiota dysbiosis by reducing the content of Proteobacteria, while increasing the content of Lactobacillus. In contrast, the effect of BL supplementation during infestation on the improvement of CR-induced colitis was not as good as that of pretreatment with BL. In conclusion, BL protects against CR-caused colitis in a preventive manner.

Keywords: Citrobacter rodentium; barley leaf; colitis; gut microbiota; prophylactic effect.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Effect of BL supplementation on mice before and during CR infestation. (A) Study design of mice grouping and treatment (n = 6 per group). (B) Weight loss. (C) Disease activity index (DAI) scores. (D) Representative images showing the gross appearance of the intestinal tissues; (E) Colon length. (F) Spleen weight. The a, b means in the same bar without a common letter differ at p < 0.05.
Figure 2
Figure 2
Effect of the beCR and duCR intervention on improving colonic histopathology. (A) Representative H&E staining images of colon tissues. Scale bar, 100 μm. Representative Alcian blue staining images of colonic sections. Scale bar, 100 μm. Representative Ki67-stained immunofluorescence images of colon tissues. Ki67 immunofluorescence is indicated in red, and DAPI nuclear staining is blue. Scale bar, 50 μm. (B) Mucosal thickness. (C) Histopathology score of colonic section. (D) Goblet cell positive density. (E) Quantification of Ki67 positive cells. The a, b, c means in the same bar without a common letter differ at p < 0.05.
Figure 2
Figure 2
Effect of the beCR and duCR intervention on improving colonic histopathology. (A) Representative H&E staining images of colon tissues. Scale bar, 100 μm. Representative Alcian blue staining images of colonic sections. Scale bar, 100 μm. Representative Ki67-stained immunofluorescence images of colon tissues. Ki67 immunofluorescence is indicated in red, and DAPI nuclear staining is blue. Scale bar, 50 μm. (B) Mucosal thickness. (C) Histopathology score of colonic section. (D) Goblet cell positive density. (E) Quantification of Ki67 positive cells. The a, b, c means in the same bar without a common letter differ at p < 0.05.
Figure 3
Figure 3
Effect of the beCR and duCR intervention on inflammatory cytokines in colon. The concentrations of anti-inflammatory cytokines (IL-4) (A) and pro-inflammatory cytokines IL-1β (B), TNF-α (C), and IFNγ (D) in colonic tissue of mice. The a, b means in the same bar without a common letter differ at p < 0.05.
Figure 3
Figure 3
Effect of the beCR and duCR intervention on inflammatory cytokines in colon. The concentrations of anti-inflammatory cytokines (IL-4) (A) and pro-inflammatory cytokines IL-1β (B), TNF-α (C), and IFNγ (D) in colonic tissue of mice. The a, b means in the same bar without a common letter differ at p < 0.05.
Figure 4
Figure 4
Effect of the beCR and duCR intervention on CR colonization. (A) Number of CR in feces on day 1, 4, 7, and 10 p.i. (B) Number of CR in spleen on day 10 p.i. (C) Quantification of CR in liver on day 10 p.i. The a, b means in the same bar without a common letter differ at p < 0.05.
Figure 5
Figure 5
Effect of the beCR and duCR intervention on modulating alpha and beta diversity of intestinal flora. The richness of gut microbiota evaluated by (A) ACE index and (B) Chao index. The diversity of gut microbiota evaluated by (C) Shannon index and (D) Simpson index. (E) NMDS analysis on OTU level. (F) PLS-DA analysis on OTU level. The a, b means in the same bar without a common letter differ at p < 0.05.
Figure 5
Figure 5
Effect of the beCR and duCR intervention on modulating alpha and beta diversity of intestinal flora. The richness of gut microbiota evaluated by (A) ACE index and (B) Chao index. The diversity of gut microbiota evaluated by (C) Shannon index and (D) Simpson index. (E) NMDS analysis on OTU level. (F) PLS-DA analysis on OTU level. The a, b means in the same bar without a common letter differ at p < 0.05.
Figure 6
Figure 6
Effect of the beCR and duCR intervention on regulating taxonomic microbial community profiles. (A) Taxonomic distributions of gut bacterial composition at the phylum level. (B) Relative abundance of Firmicutes and Proteobacteria. (C) Relative abundance of Citrobacter, Lactobacillus, unclassified_f_Lachnospiraceae, norank_f_Lachnospiraceae, Enterorhabdus, and Erysipelatoclostridium. (D) Wilcoxon rank-sum test of group CD+CR and duCR at the genus level. (E) Wilcoxon rank-sum test of group CD+CR and beCR at the genus level. (F) Spearman correlations analysis between the microbiota and colitis-related index. The a, b means in the same bar without a common letter differ at p < 0.05.
Figure 7
Figure 7
Effect of the beCR and duCR intervention on key microbial phylotypes. (A) Cladogram and (B) LDA scores derived from LEfSe analysis.
Figure 8
Figure 8
Schematic diagram of BLs’ role in improving CR-causing colitis through a preventive approach.
See this image and copyright information in PMC

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