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Human umbilical cord-derived mesenchymal stem cells ameliorate experimental colitis by normalizing the gut microbiota - PubMed

  • ️Sat Jan 01 2022

Human umbilical cord-derived mesenchymal stem cells ameliorate experimental colitis by normalizing the gut microbiota

Fan Yang et al. Stem Cell Res Ther. 2022.

Abstract

Background: Crohn's disease (CD) is a chronic non-specific inflammatory bowel disease. Current CD therapeutics cannot fundamentally change the natural course of CD. Therefore, it is of great significance to find new treatment strategies for CD. Preclinical and clinical studies have shown that mesenchymal stromal cells (MSCs) are a promising therapeutic approach. However, the mechanism by which MSCs alleviate CD and how MSCs affect gut microbes are still unclear and need further elucidation.

Methods: We used 2,4,6-trinitrobenzenesulfonic acid (TNBS) to induce experimental colitis in mice and analysed the microbiota in faecal samples from the control group, the TNBS group and the TNBS + MSC group with faecal 16S rDNA sequencing. Subsequent analyses of alpha and beta diversity were all performed based on the rarified data. PICRUStII analysis was performed on the 16S rRNA gene sequences to infer the gut microbiome functions.

Results: MSC Treatment improved TNBS-induced colitis by increasing survival rates and relieving symptoms. A distinct bacterial signature was found in the TNBS group that differed from the TNBS + MSC group and controls. MSCs prevented gut microbiota dysbiosis, including increasing α-diversity and the amount of Bacteroidetes Firmicutes and Tenericutes at the phylum level and decreasing the amount of Proteobacteria at the phylum level. MSCs alleviated the increased activities of sulphur and riboflavin metabolism. Meanwhile some metabolic pathways such as biosynthesis of amino acids lysine biosynthesis sphingolipid metabolism and secondary bile acid biosynthesis were decreased in the TNBS group compared with the control group and the TNBS + MSC group CONCLUSIONS: Overall, our findings preliminarily confirmed that colitis in mice is closely related to microbial and metabolic dysbiosis. MSC treatment could modulate the dysregulated metabolism pathways in mice with colitis, restoring the abnormal microbiota function to that of the normal control group. This study provides insight into specific intestinal microbiota and metabolism pathways linked with MSC treatment, suggesting a new approach to the treatment of CD.

Keywords: 16S rRNA gene sequences; Crohn's disease; Gut microbiota; Mesenchymal stem cells; Metabolism; TNBS.

© 2022. The Author(s).

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1

Human umbilical cord-derived mesenchymal stem cell (hUC-MSC) therapy mitigates TNBS-induced colitis. Colitis BALB/c mice were treated with hUC-MSCs (106 per mouse), while the control mice received only control medium (saline) 2 h following rectal administration of 2,4,6-trinitrobenzenesulfonic acid (TNBS). The animals were monitored every day for a total of 7 days for weight loss, the consistency of their stool, the presence of blood in their stool, disease activity and histologic scores, and the expression of mucosal barrier proteins. a Decreased weight loss; b alleviated colitis symptoms; c, d Colons were examined for their general form and length 3 days after TNBS intracolonic administration; e Histopathologic analysis (H&E staining and histological score). Inflammation was graded from 0 to 4; f Expression of the tight junction-related proteins occludin and ZO-1. The stained sections were read under a microscope and quantified using Image-Pro Plus (IPP) 6.0 software. The quantified results of occludin(g) and ZO-1(h) are presented as the mean density. The mean densities of 5 randomly selected fields in each group. Data are shown as the mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, and n.s. means not significant

Fig. 2
Fig. 2

Changes in faecal microbial diversities among the control group (n = 4), TNBS group (n = 5) and TNBS + MSC group (n = 5). Faecal pellets were collected 3 days after MSC injection from the control TNBS and TNBS + MSC groups, and 16SrRNA sequences were determined as described in the "Methods". Alpha-diversity analysis and beta-diversity analysis among the three groups. a Pan genome analysis, b core genome analysis, c rank abundance curve of bacterial OTUs among the three groups. d–f The Sob, Shannon and Shannoneven indices were used to estimate the diversity of the faecal microbiota among the three groups (data expressed as the mean ± SD). g The plots shown were generated using principal coordinate analysis (PCoA). h Adonis shows that the difference between groups is significantly greater than that within groups (P < 0.001). i The Venn plot can be used to count the number of species that are common and unique among the three groups

Fig. 3
Fig. 3

Structural comparison of the faecal microbiota among the control group (n = 4), TNBS group (n = 5) and TNBS + MSC group (n = 5). a Clustering of bacterial microbial composition at the phylum level in different samples. b The bacterial microbial composition in different experimental groups at the phylum levels. c The relative abundance of bacterial groups at the phylum level between groups tested by means of one-way nonparametric Analysis of Variance (ANOVA) (Kruskal–Wallis H test). *P < 0.05, **P < 0.01, ***P < 0.001. d Clustering of the bacterial microbial composition at the genus level in different samples. e The composition of bacterial microbial composition in different experimental groups at the genus level. f The relative abundance of bacterial groups at the genus level between groups tested by one-way nonparametric ANOVA(Kruskal–Wallis H test). *P < 0.05, **P < 0.01, ***P < 0.001

Fig. 4
Fig. 4

Differences in dominant microorganisms between the control and TNBS groups. In the comparisons of the control group versus the TNBS groups, colitis mice were enriched in the Proteobacteria phylum based on the LDA scores and cladogram. a LDA scores from LefSe analysis were performed on relative OTU abundances. Least discriminant analysis (LDA) effect size taxonomic cladogram comparing all samples categorized by control (red) and TNBS groups (blue). b Distribution histogram based on LDA (LDA score > 3). Significantly discriminant taxon nodes are coloured, and the branch areas are shaded according to the highest-ranked variety for that taxon. For each taxon detected, the corresponding node in the taxonomic cladogram is coloured according to the highest-ranked group for that taxon. If the taxon is not significantly differentially represented between sample groups, the corresponding node is coloured yellow

Fig. 5
Fig. 5

Differences in dominant microorganisms between the TNBS and TNBS + MSCgroups. The TNBS group had higher enrichment of the Proteobacteria phylum, while the TNBS + MSC group had higher enrichment of the Firmicutes phylum a LDA scores from LefSe analysis were obtained from the relative OTU abundances and the least discriminant analysis (LDA) effect size taxonomic cladogram comparing all samples categorized by TNBS (red) and the TNBS + MSC group (blue). b Distribution histogram based on LDA (LDA score > 3). Significantly discriminant taxon nodes are coloured and the branch areas are shaded according to the highest-ranked variety for that taxon. For each taxon detected, the corresponding node in the taxonomic cladogram is coloured according to the highest-ranked group for that taxon. If the taxon is not significantly differentially represented between the sample groups, the corresponding node is depicted in yellow

Fig. 6
Fig. 6

Gut microbiome functions inferred by PICRUStII from 16S rRNA gene sequences among the three groups (the control group, the TNBS group and the TNBS + MSC group). a The blue, golden yellow and green bar graphs indicate the inferred metabolism value of control group, TNBS group and TNBS + MSC group. KEGG orthology was used for functional categorization. STAMP3 was used for functional profiling. Elevated metabolic pathways after modelling. b Decreased metabolic pathways after modelling. A box plot shows the top quartile, median, and bottom quartile, while white stars indicate the average and a plus sign indicates the outlier. All differences were analysed using one-way ANOVA followed by the Tukey–Kramer post hoc test. The multiple comparisons were not corrected

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