Ileal Bile Acid Transporter Inhibitor Improves Hepatic Steatosis by Ameliorating Gut Microbiota Dysbiosis in NAFLD Model Mice

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

study design

laboratory experiment

Citation

PMID PubMed identifier for scientific articles.

34225483

DOI Digital object identifier for electronic documents.

10.1128/mBio.01155-21

URI

Authors

Matsui M, Fukunishi S, Nakano T, Ueno T, Higuchi K, Asai A

Journal

mBio

Year

2021

Keywords:

gut microbiome, ileal bile acid transporter inhibitor, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis

Nonalcoholic fatty liver disease (NAFLD), characterized by excessive fat deposition in the liver unrelated to alcohol consumption, is highly prevalent worldwide. However, effective therapeutic agents approved for NAFLD treatment are lacking. An ileal bile acid transporter inhibitor (IBATi), which represents a new mode of treatment of chronic idiopathic constipation, leads to increased delivery of bile acids to the colon. We investigated the effect of IBATi against NAFLD through modification of the gut microbiota in mice. IBATi treatment significantly suppressed body weight gain, liver dysfunction, and serum low-density lipoprotein levels and significantly decreased NAFLD activity scores in high-fat diet (HFD) mice. Treatment with IBATi ameliorated the decreased hepatic cholesterol 7-a-monooxygenase (Cyp7a1) and increased ileal fibroblast growth factor 15 (Fgf15) mRNA expression in HFD mice. Further, IBATi treatment changed the α-diversity in the gut microbiota reduced by HFD, which was analyzed in feces using 16S rRNA sequencing. To establish the mechanism underlying improvement in NAFLD induced by IBATi, we recolonized antibiotic solution-treated mice by fecal microbiome transplantation (FMT) using stool from HFD or HFD plus IBATi mice. This is the first report that fecally transplanted gut microbiota from HFD plus IBATi mice prevented hepatic steatosis caused by HFD. In conclusion, IBATi improved hepatic steatosis by ameliorating gut microbiota dysbiosis in NAFLD model mice, suggesting a potential therapeutic agent for NAFLD treatment. IMPORTANCE NAFLD is an increasingly recognized condition that may progress to liver cirrhosis and hepatocellular carcinoma, and community surveys have assessed that the prevalence is 14 to 32% worldwide. The first line of treatment for NAFLD is lifestyle modification to achieve weight reduction, particularly through diet and exercise. However, weight reduction is difficult to achieve and maintain, and pharmacological agents approved for the treatment of NAFLD are lacking. This study investigated the influence of the gut microbiota and the effect of an IBATi on NAFLD using a murine model. Treatment with IBATi significantly improved NAFLD in HFD mice. Further, fecal microbiome transplantation using stool from HFD plus IBATi mice prevented hepatic steatosis caused by HFD. Our study makes a significant contribution to the literature because the study findings suggest a potential treatment strategy for NAFLD patients by ameliorating gut microbiota dysbiosis.

Experiment 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/13

Curator: Tosin

Revision editor(s): Tosin

Subjects

Location of subjects

Japan

Host species Species from which microbiome was sampled. Contact us to have more species added.

Mus musculus

Body site Anatomical site where microbial samples were extracted from according to the Uber Anatomy Ontology

Feces Cow dung,Cow pat,Droppings,Dung,Excrement,Excreta,Faeces,Fecal material,Fecal matter,Fewmet,Frass,Guano,Matières fécales@fr,Merde@fr,Ordure,Partie de la merde@fr,Piece of shit,Porción de mierda@es,Portion of dung,Portion of excrement,Portion of faeces,Portion of fecal material,Portion of fecal matter,Portion of feces,Portion of guano,Portion of scat,Portionem cacas,Scat,Spoor,Spraint,Stool,Teil der fäkalien@de,Feces,feces

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology

Treatment outcome measurement Treatment outcome measurement,treatment outcome measurement

Group 0 name Corresponds to the control (unexposed) group for case-control studies

Combination of IBATi (ileal bile acid transporter inhibitor), HFD (High fat diet) and HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) groups

Group 1 name Corresponds to the case (exposed) group for case-control studies

Control diet

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Mice in the control diet groups were subjected to the same oral dose of phosphate-buffered saline (PBS).

Group 0 sample size Number of subjects in the control (unexposed) group

9

Group 1 sample size Number of subjects in the case (exposed) group

3

Lab analysis

Sequencing type

16S

16S variable region One or more hypervariable region(s) of the bacterial 16S gene

V3-V4

Sequencing platform Manufacturer and experimental platform used for quantifying microbial abundance

Illumina

Statistical Analysis

Data transformation Data transformation applied to microbial abundance measurements prior to differential abundance testing (if any).

relative abundances

Statistical test

LEfSe

Significance threshold p-value or FDR threshold used for differential abundance testing (if any)

0.05

MHT correction Have statistical tests be corrected for multiple hypothesis testing (MHT)?

No

LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool

2

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Curated date: 2025/04/14

Curator: Tosin

Revision editor(s): Tosin

Source: Figure 1D, Figure S3A and Figure S3B

Description: Relative abundance at the family, genus and species level by linear discriminant analysis effect size (LEfSe) in each group

Abundance in Group 1: increased abundance in Control diet

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Cytophagaceae
Flammeovirgaceae
Flavobacteriaceae
Lachnoclostridium
Parabacteroides distasonis
Phocaeicola sartorii
Porphyromonadaceae
Prevotellaceae
Staphylococcaceae
Sutterellaceae
Lachnoclostridium indolisLachnoclostridium indolis

Revision editor(s): Tosin

Experiment 2

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Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

Combination of Control diet, HFD (High fat diet) and HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) groups

Group 1 name Corresponds to the case (exposed) group for case-control studies

IBATi (ileal bile acid transporter inhibitor) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Mice treated with IBATi (ileal bile acid transporter inhibitor)

Lab analysis

Statistical Analysis

Signature 1

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Curator: Tosin

Revision editor(s): Tosin

Source: Figure 1D, Figure S3A and Figure S3B

Description: Relative abundance at the family, genus and species level by linear discriminant analysis effect size (LEfSe) in each group

Abundance in Group 1: increased abundance in IBATi (ileal bile acid transporter inhibitor) group

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Erysipelotrichaceae
Lactobacillaceae
Lactobacillus
Alistipes
Eubacterium
Limosilactobacillus reuteri
Lactobacillus johnsonii
Alistipes putredinis

Revision editor(s): Tosin

Experiment 3

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/14

Curator: Tosin

Revision editor(s): Tosin

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

Combination of Control diet, IBATi (ileal bile acid transporter inhibitor) and HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) groups

Group 1 name Corresponds to the case (exposed) group for case-control studies

HFD (High fat diet) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Six weeks Old mice fed with High fat diet for 12 weeks

Lab analysis

Statistical Analysis

Signature 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/14

Curator: Tosin

Revision editor(s): Tosin

Source: Figure 1D, Figure S3A and Figure S3B

Description: Relative abundance at the family, genus and species level by linear discriminant analysis effect size (LEfSe) in each group

Abundance in Group 1: increased abundance in HFD (High fat diet) group

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Eubacteriaceae
Streptococcaceae
Bacteroides sp.
Lactococcus lactis
Lactococcus
Dorea
Roseburia
Tyzzerella

Revision editor(s): Tosin

Experiment 4

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/14

Curator: Tosin

Revision editor(s): Tosin

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

Combination of Control diet, IBATi (ileal bile acid transporter inhibitor) and HFD (High fat diet) groups

Group 1 name Corresponds to the case (exposed) group for case-control studies

HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Mice fed with High fat diet and ileal bile acid transporter inhibitor

Lab analysis

Statistical Analysis

Signature 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/14

Curator: Tosin

Revision editor(s): Tosin

Source: Figure 1D and Figure S3B

Description: Relative abundance at the family and species level by linear discriminant analysis effect size (LEfSe) in each group

Abundance in Group 1: increased abundance in HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) group

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Desulfovibrionaceae
Clostridium sp.

Revision editor(s): Tosin

Experiment 5

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Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology

Response to transplant Response to transplant,response to transplant

Group 0 name Corresponds to the control (unexposed) group for case-control studies

Mice with FMT (fecal microbiota transplantation) from HFD (high fat diet) mice

Group 1 name Corresponds to the case (exposed) group for case-control studies

Mice with FMT (fecal microbiota transplantation) from HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) mice

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Mice with FMT (fecal microbiota transplantation) from HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) mice

Group 0 sample size Number of subjects in the control (unexposed) group

3

Lab analysis

Statistical Analysis

Statistical test

T-Test

LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool

Not specified

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness

increased

Signature 1

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Curated date: 2025/04/14

Curator: Tosin

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Source: Figure 4C

Description: Microbial composition at the family level in each FMT group.

Abundance in Group 1: increased abundance in Mice with FMT (fecal microbiota transplantation) from HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) mice

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Porphyromonadaceae
Eubacteriaceae

Revision editor(s): Tosin

Signature 2

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Curated date: 2025/04/14

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Source: Figure 4C

Description: Microbial composition at the family level in each FMT group.

Abundance in Group 1: decreased abundance in Mice with FMT (fecal microbiota transplantation) from HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) mice

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Bacteroidaceae
Streptococcaceae

Revision editor(s): Tosin

Experiment 6

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/14

Curator: Tosin

Revision editor(s): Tosin

Differences from previous experiment shown

Subjects

Lab analysis

Statistical Analysis

Statistical test

LEfSe

LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool

2

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness

increased

Signature 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/14

Curator: Tosin

Revision editor(s): Tosin

Source: Figure 4D, Figure S3C and S3D

Description: Relative abundance at the family, genus and species level by LEfSe in each FMT group.

Abundance in Group 1: increased abundance in Mice with FMT (fecal microbiota transplantation) from HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) mice

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Acetatifactor muris
Adlercreutzia caecimuris
Alistipes
Bacillaceae
Blautia
Clostridium
Cytophagaceae
Erysipelotrichaceae
Eubacteriaceae
Faecalibacterium
Kineosporiaceae
Mediterraneibacter gnavus
Oscillibacter
Parabacteroides
Parabacteroides goldsteinii
Porphyromonadaceae
Proteus
Rikenellaceae
Ruminococcus
Streptococcus
Streptococcus danieliae
Streptomycetaceae
Syntrophomonadaceae
Thomasclavelia
Thomasclavelia cocleata
Erysipelatoclostridium cocleatumErysipelatoclostridium cocleatum

Revision editor(s): Tosin

Signature 2

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/14

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Source: Figure 4D, Figure S3C and S3D

Description: Relative abundance at the family, genus and species level by LEfSe in each FMT group.

Abundance in Group 1: decreased abundance in Mice with FMT (fecal microbiota transplantation) from HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) mice

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Anaerotruncus
Bacteroidaceae
Bacteroides
Bacteroides acidifaciens
Bacteroides sp.
Bacteroides xylanisolvens
Deferribacteraceae
Desulfovibrio
Desulfovibrionaceae
Dorea
Dorea formicigenerans
Ligilactobacillus animalis
Ligilactobacillus murinus
Mucispirillum
Mucispirillum schaedleri
Parabacteroides distasonis
Parvibacter
Roseburia
Roseburia faecis
Roseburia intestinalis
Thermoanaerobacteraceae
Parvibacter distasonisParvibacter distasonis
Desulfovibrio C21 c20Desulfovibrio C21 c20

Revision editor(s): Tosin

Experiment 7

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin

Differences from previous experiment shown

Subjects

Condition The experimental condition / phenotype studied according to the Experimental Factor Ontology

Treatment outcome measurement Treatment outcome measurement,treatment outcome measurement

Group 0 name Corresponds to the control (unexposed) group for case-control studies

Control diet group

Group 1 name Corresponds to the case (exposed) group for case-control studies

HFD (High fat diet) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Mice fed with High fat diet

Lab analysis

Statistical Analysis

Statistical test

T-Test

LDA Score above Threshold for the linear discriminant analysis (LDA) score for studies using the popular LEfSe tool

Not specified

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness

decreased

Signature 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin

Source: Figure 1C

Description: Microbial composition at the family level in each group

Abundance in Group 1: increased abundance in HFD (High fat diet) group

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Streptococcaceae
Desulfovibrionaceae
Eubacteriaceae

Revision editor(s): Tosin

Signature 2

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin

Source: Figure 1C

Description: Microbial composition at the family level in each group

Abundance in Group 1: decreased abundance in HFD (High fat diet) group

NCBI	Quality Control	Links
Porphyromonadaceae
Prevotellaceae
Flavobacteriaceae

Revision editor(s): Tosin

Experiment 8

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin

Differences from previous experiment shown

Subjects

Group 1 name Corresponds to the case (exposed) group for case-control studies

HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Mice fed with High fat diet and ileal bile acid transporter inhibitor

Lab analysis

Statistical Analysis

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness

unchanged

Signature 1

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Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin, KateRasheed

Source: Figure 1C

Description: Microbial composition at the family level in each group

Abundance in Group 1: increased abundance in HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) group

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Clostridiaceae
Geobacteraceae
Lachnospiraceae
Streptococcaceae

Revision editor(s): Tosin, KateRasheed

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin, KateRasheed

Source: Figure 1C

Description: Microbial composition at the family level in each group

Abundance in Group 1: decreased abundance in HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) group

NCBI	Quality Control	Links
Flavobacteriaceae
Porphyromonadaceae
Prevotellaceae

Revision editor(s): Tosin, KateRasheed

Experiment 9

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin

Differences from previous experiment shown

Subjects

Group 1 name Corresponds to the case (exposed) group for case-control studies

IBATi (ileal bile acid transporter inhibitor) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Mice treated with IBATi (ileal bile acid transporter inhibitor)

Lab analysis

Statistical Analysis

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness

unchanged

Signature 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin

Source: Figure 1C

Description: Microbial composition at the family level in each group

Abundance in Group 1: increased abundance in IBATi (ileal bile acid transporter inhibitor) group

NCBI	Quality Control	Links
Rikenellaceae

Revision editor(s): Tosin

Signature 2

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin

Source: Figure 1C

Description: Microbial composition at the family level in each group

Abundance in Group 1: decreased abundance in IBATi (ileal bile acid transporter inhibitor) group

NCBI	Quality Control	Links
Flavobacteriaceae
Prevotellaceae

Revision editor(s): Tosin

Experiment 10

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

HFD (High fat diet) groups

Group 1 name Corresponds to the case (exposed) group for case-control studies

HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) group

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Mice fed with High fat diet and ileal bile acid transporter inhibitor

Lab analysis

Statistical Analysis

Signature 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Tosin

Revision editor(s): Tosin

Source: Figure 1C

Description: Microbial composition at the family level in each group

Abundance in Group 1: decreased abundance in HFD + IBATi (High fat diet + ileal bile acid transporter inhibitor) group

NCBI	Quality Control	Links
Porphyromonadaceae
Streptococcaceae

Revision editor(s): Tosin

Experiment 11

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Anne-mariesharp

Revision editor(s): Anne-mariesharp

Differences from previous experiment shown

Subjects

Group 0 name Corresponds to the control (unexposed) group for case-control studies

Control

Group 1 name Corresponds to the case (exposed) group for case-control studies

IBATi -Tx (Ileal Bile Acid Transporter inhibitor - Treatment)

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Six-week-old mice were fed HFD(High Fat Diet) for 12 weeks and received IBATi (Ileal Bile Acid Transporter inhibitor) treatment between weeks 6 to 12.

Lab analysis

Statistical Analysis

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness

increased

Signature 1

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Anne-mariesharp

Revision editor(s): Anne-mariesharp

Source: Fig. 8C

Description: Microbial composition at the family level in both groups.

Abundance in Group 1: increased abundance in IBATi -Tx (Ileal Bile Acid Transporter inhibitor - Treatment)

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Porphyromonadaceae

Revision editor(s): Anne-mariesharp

Signature 2

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Curator: Anne-mariesharp

Revision editor(s): Anne-mariesharp

Source: Fig. 8C

Description: Microbial composition at the family level in both groups.

Abundance in Group 1: decreased abundance in IBATi -Tx (Ileal Bile Acid Transporter inhibitor - Treatment)

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Lachnospiraceae
Streptococcaceae

Revision editor(s): Anne-mariesharp

Experiment 12

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Anne-mariesharp

Revision editor(s): Anne-mariesharp

Differences from previous experiment shown

Subjects

Group 1 definition Diagnostic criteria applied to define the specific condition / phenotype represented in the case (exposed) group

Six-week-old mice were fed HFD(High Fat Diet) for 12 weeks and received IBATi (Ileal Bile Acid Transporter inhibitor) treatment between weeks 6 to 12.

Lab analysis

Statistical Analysis

Statistical test

LEfSe

Alpha Diversity

Shannon Estimator of species richness and species evenness: more weight on species richness

increased

Signature 1

EditHistoryDelete

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Anne-mariesharp

Revision editor(s): Anne-mariesharp

Source: Fig. 8D, Supplemental figure 3E & 3F

Description: Microbial composition at the family, genus and species level in both groups.

Abundance in Group 1: increased abundance in IBATi -Tx (Ileal Bile Acid Transporter inhibitor - Treatment)

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Atopobium
Bacteroides acidifaciens
Coriobacteriaceae
Cyclobacteriaceae
Flammeovirgaceae
Flavobacteriaceae
Oscillibacter
Porphyromonadaceae
Rikenellaceae
Streptomycetaceae
Clostridium sp.
Lactobacillus johnsonii
Peptacetobacter hiranonis

Revision editor(s): Anne-mariesharp

Signature 2

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Reviewed Marked as Reviewed by KateRasheed on 2025-4-28

Curated date: 2025/04/15

Curator: Anne-mariesharp

Revision editor(s): Anne-mariesharp

Source: Fig. 8D, Supplemental figure 3E & 3F

Description: Microbial composition at the family, genus and species level in both groups.

Abundance in Group 1: decreased abundance in IBATi -Tx (Ileal Bile Acid Transporter inhibitor - Treatment)

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Anaerotruncus colihominis
Blautia
Blautia producta
Dorea
Erysipelotrichaceae
Lactococcus
Lactococcus lactis
Marvinbryantia
Mediterraneibacter gnavus
Oscillibacter valericigenes
Roseburia
Ruminococcus
Streptococcaceae
Streptococcus
Veillonellaceae
Lachnoclostridium indolisLachnoclostridium indolis

Revision editor(s): Anne-mariesharp