CoChat
Sign Up
Cited by 405Gut microbiota and healthBacterial biofilms and quorum sensingGastrointestinal motility and disorders

The DOI matches its Crossref record; 41 of the 41 references it cites were found in a registry or an index.

Commensal Bacteria-Dependent Indole Production Enhances Epithelial Barrier Function in the Colon

Open access
Shimada, Yosuke, Kinoshita, Makoto, Harada, Kazuo, Mizutani, Masafumi, Masahata, Kazunori, Kayama, Hisako, Takeda, Kiyoshi·2013·PLoS ONE
DOI 10.1371/journal.pone.0080604PMC PMC3835565
Open this paper in CoChat — free accountOpen PDF

Abstract

Microbiota have been shown to have a great influence on functions of intestinal epithelial cells (ECs). The role of indole as a quorum-sensing (QS) molecule mediating intercellular signals in bacteria has been well appreciated. However, it remains unknown whether indole has beneficial effects on maintaining intestinal barriers in vivo. In this study, we analyzed the effect of indole on ECs using a germ free (GF) mouse model. GF mice showed decreased expression of junctional complex molecules in colonic ECs. The feces of specific pathogen-free (SPF) mice contained a high amount of indole; however the amount was significantly decreased in the feces of GF mice by 27-fold. Oral administration of indole-containing capsules resulted in increased expression of both tight junction (TJ)- and adherens junction (AJ)-associated molecules in colonic ECs in GF mice. In accordance with the increased expression of these junctional complex molecules, GF mice given indole-containing capsules showed higher resistance to dextran sodium sulfate (DSS)-induced colitis. A similar protective effect of indole on DSS-induced epithelial damage was also observed in mice bred in SPF conditions. These findings highlight the beneficial role of indole in establishing an epithelial barrier in vivo.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0080604

Cites

41 of 41 references verified0 citation errors · 0 not checkable

Statistics

Figures

Figure 1 Epithelial barrier functions is impaired in GF mice. (A) Real-time quantitative RT-PCR analysis of mRNA expression of Cldn7, Ocln, Tjp1, Ctnnb1, Cdh1 in colonic ECs in SPF (n = 4) or GF (n = 4) mice. Values were normalized to that of Gapdh. Data are representative of two independent experiments and show mean values ± S.D. of 4 samples performed in duplicate. *P<0.05. (B) Mouse colonic tissue was stained with anti-occludin antibody. Sections were analyzed using a confocal microscope. Bars, 50 µm. Data are representative of two independent experiments. (C) SPF (n = 8) or GF (n = 8) mice were administered 4% DSS by drinking water for 3 days. Survival rates of the indicated mice are shown. Body weight changes relative to the value prior to colitis induction are shown. Data are mean ± S.E.M of 8 mice at each time point. SPF, specific pathogen free; GF, germ free.
Figure 1 Epithelial barrier functions is impaired in GF mice. (A) Real-time quantitative RT-PCR analysis of mRNA expression of Cldn7, Ocln, Tjp1, Ctnnb1, Cdh1 in colonic ECs in SPF (n = 4) or GF (n = 4) mice. Values were normalized to that of Gapdh. Data are representative of two independent experiments and show mean values ± S.D. of 4 samples performed in duplicate. *P<0.05. (B) Mouse colonic tissue was stained with anti-occludin antibody. Sections were analyzed using a confocal microscope. Bars, 50 µm. Data are representative of two independent experiments. (C) SPF (n = 8) or GF (n = 8) mice were administered 4% DSS by drinking water for 3 days. Survival rates of the indicated mice are shown. Body weight changes relative to the value prior to colitis induction are shown. Data are mean ± S.E.M of 8 mice at each time point. SPF, specific pathogen free; GF, germ free.CC BY · PubMed Central
Figure 2 Indole and indole metabolites are absent in GF mice. (A, B) Feces and serum were collected from either SPF (n = 3) or GF (n = 3) mice. The concentration of indole in the feces was measured by HPLC-FL, and the serum concentration of indoxyl sulfate was measured by LC-MS/MS. Data are representative of two independent experiments and show mean values ± S.D. of 3 mice. *P<0.05. SPF, specific pathogen free; GF, germ free.
Figure 2 Indole and indole metabolites are absent in GF mice. (A, B) Feces and serum were collected from either SPF (n = 3) or GF (n = 3) mice. The concentration of indole in the feces was measured by HPLC-FL, and the serum concentration of indoxyl sulfate was measured by LC-MS/MS. Data are representative of two independent experiments and show mean values ± S.D. of 3 mice. *P<0.05. SPF, specific pathogen free; GF, germ free.CC BY · PubMed Central
Figure 3 Indole, but not indoxyl sulfate, induces the expression of junctional complex molecules in Caco-2 cells. (A, B) Real-time quantitative RT-PCR analysis of mRNA expression of Cldn7, Ocln, Tjp1, Ctnnb1, Cdh1 in Caco-2 cells cultured with indole or indoxyl sulfate is shown. DMF or PBS was used as a control, respectively. Quadruplicate was used for each condition. Values were normalized to the expression of Gapdh. Data are representative of two independent experiments and show mean values ± S.D. of 4 samples performed in duplicate. *P<0.05. n.s., not significant.
Figure 3 Indole, but not indoxyl sulfate, induces the expression of junctional complex molecules in Caco-2 cells. (A, B) Real-time quantitative RT-PCR analysis of mRNA expression of Cldn7, Ocln, Tjp1, Ctnnb1, Cdh1 in Caco-2 cells cultured with indole or indoxyl sulfate is shown. DMF or PBS was used as a control, respectively. Quadruplicate was used for each condition. Values were normalized to the expression of Gapdh. Data are representative of two independent experiments and show mean values ± S.D. of 4 samples performed in duplicate. *P<0.05. n.s., not significant.CC BY · PubMed Central
Figure 4 Indole-containing capsules promote epithelial barrier function in GF mice. (A) Feces were collected from SPF mice, and GF mice treated with indole- or MCT- containing capsules. Three mice was analysed in each group. The concentration of indole in the feces was measured by HPLC-FL. Data show mean values ± S.D. of 3 samples. *P<0.05. n.s., not significant. SPF, specific pathogen free; GF, germ free; MCT, Medium-Chain Triglycerides. (B) Real-time quantitative RT-PCR analysis of mRNA expression of Cldn7, Ocln, Tjp1, Ctnnb1, and Cdh1 in colonic epithelial cells of GF mice treated with indole- (n = 4) or MCT- (n = 4) containing capsules. Values were normalized to the expression of Gapdh. Data are representative of two independent experiments and show mean values ± S.D. of 4 samples performed in duplicate. *P<0.05. (C) Colonic tissues of GF mice treated with indole- or MCT- containing capsules were stained with anti-occludin antibody. Sections were analyzed using a confocal microscope. Bars, 20 µm. Data are representative of two independent experiments. (D) After oral administration with either indole- (n = 6) or MCT- (n = 6) containing capsules for 2 weeks, GF mice were treated by 4% DSS in drinking water for 3 days. Survival rate of the mice in each group is shown. Data are representative of two independent experiments. MCT, Medium-Chain Triglycerides.
Figure 4 Indole-containing capsules promote epithelial barrier function in GF mice. (A) Feces were collected from SPF mice, and GF mice treated with indole- or MCT- containing capsules. Three mice was analysed in each group. The concentration of indole in the feces was measured by HPLC-FL. Data show mean values ± S.D. of 3 samples. *P<0.05. n.s., not significant. SPF, specific pathogen free; GF, germ free; MCT, Medium-Chain Triglycerides. (B) Real-time quantitative RT-PCR analysis of mRNA expression of Cldn7, Ocln, Tjp1, Ctnnb1, and Cdh1 in colonic epithelial cells of GF mice treated with indole- (n = 4) or MCT- (n = 4) containing capsules. Values were normalized to the expression of Gapdh. Data are representative of two independent experiments and show mean values ± S.D. of 4 samples performed in duplicate. *P<0.05. (C) Colonic tissues of GF mice treated with indole- or MCT- containing capsules were stained with anti-occludin antibody. Sections were analyzed using a confocal microscope. Bars, 20 µm. Data are representative of two independent experiments. (D) After oral administration with either indole- (n = 6) or MCT- (n = 6) containing capsules for 2 weeks, GF mice were treated by 4% DSS in drinking water for 3 days. Survival rate of the mice in each group is shown. Data are representative of two independent experiments. MCT, Medium-Chain Triglycerides.CC BY · PubMed Central
Figure 5 Indole-containing capsules show preventative effect on colitis development in SPF mice. SPF mice were treated with indole- (n = 7) or MCT- (n = 7) containing capsules for 1 week, and then challenged by 5% DSS for 6 days. Body weight changes relative to the value prior to colitis induction are shown. Data are representative of two independent experiments and mean ± S.E.M of 7 mice at each time point is shown. *P<0.05. MCT, Medium-Chain Triglycerides.
Figure 5 Indole-containing capsules show preventative effect on colitis development in SPF mice. SPF mice were treated with indole- (n = 7) or MCT- (n = 7) containing capsules for 1 week, and then challenged by 5% DSS for 6 days. Body weight changes relative to the value prior to colitis induction are shown. Data are representative of two independent experiments and mean ± S.E.M of 7 mice at each time point is shown. *P<0.05. MCT, Medium-Chain Triglycerides.CC BY · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Show all 41 references

Cited by

Sorted by citations

See all 405

Related

Metadata

Authors
Shimada, Y., Kinoshita, M., Harada, K., Mizutani, M., Masahata, K., Kayama, H., & Takeda, K.
Affiliations
  • Immunology Frontier Research Center, The University of Osaka (JP)
  • Japan Science and Technology Agency (JP)
  • The University of Osaka (JP)
  • Morishita Jintan (Japan) (JP)
Corresponding author
Takeda, Kiyoshi
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2013-11-20
Volume
8
Issue
11
Pages
e80604
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0080604
PMC
PMC3835565
OpenAlex
W2040501063
Cited by
404
References
41
Citations by year
2026: 23 · 2025: 43 · 2024: 50 · 2023: 53 · 2022: 44 · 2021: 47 · 2020: 34 · 2019: 27 · 2018: 24 · 2017: 23 · 2016: 15 · 2015: 15 · 2014: 6
FWCI
5.37
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
Funders
  • Ministry of Education, Culture, Sports, Science and Technology
  • Ministry of Health, Labour and Welfare
  • Japan Society for the Promotion of Science (24390120)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gut microbiota and health
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Bacterial biofilms and quorum sensing
  • Health Sciences › Medicine › Gastroenterology › Gastrointestinal motility and disorders
Keywords
indole, commensal bacteria, intestinal epithelial barrier, tight junctions, adherens junctions, quorum sensing, germ-free mice, DSS-induced colitis, colonic epithelial cells, gut microbiota, epithelial permeability
MeSH terms
Animals, Bacteria, Chromatography, High Pressure Liquid, Colon, Germ-Free Life, Indoles, Intestinal Mucosa, Mice, Tandem Mass Spectrometry, Real-Time Polymerase Chain Reaction

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review