Cited by 754Gut microbiota and healthSirtuins and Resveratrol in MedicineCholesterol and Lipid Metabolism

The DOI matches its Crossref record; 49 of the 50 references it cites were found in a registry or an index; 10 of the 75 sources its claims cite were checked for support.

Resveratrol Attenuates Trimethylamine- N -Oxide (TMAO)-Induced Atherosclerosis by Regulating TMAO Synthesis and Bile Acid Metabolism via Remodeling of the Gut Microbiota

Open access
Chen, Ming-liang, Yi, Long, Zhang, Yong, Zhou, Xi, Ran, Li, Yang, Jining, Zhu, Jun-dong, Zhang, Qian-yong, Mi, Man-tian·2016·mBio

Abstract

The gut microbiota is found to be strongly associated with atherosclerosis (AS). Resveratrol (RSV) is a natural phytoalexin with anti-AS effects; however, its mechanisms of action remain unclear. Therefore, we sought to determine whether the anti-AS effects of RSV were related to changes in the gut microbiota. We found that RSV attenuated trimethylamine- N -oxide (TMAO)-induced AS in ApoE −/− mice. Meanwhile, RSV decreased TMAO levels by inhibiting commensal microbial trimethylamine (TMA) production via gut microbiota remodeling in mice. Moreover, RSV increased levels of the genera Lactobacillus and Bifidobacterium , which increased the bile salt hydrolase activity, thereby enhancing bile acid (BA) deconjugation and fecal excretion in C57BL/6J and ApoE −/− mice. This was associated with a decrease in ileal BA content, repression of the enterohepatic farnesoid X receptor (FXR)-fibroblast growth factor 15 (FGF15) axis, and increased cholesterol 7a-hydroxylase (CYP7A1) expression and hepatic BA neosynthesis. An FXR antagonist had the same effect on FGF15 and CYP7A1 expression as RSV, while an FXR agonist abolished RSV-induced alterations in FGF15 and CYP7A1 expression. In mice treated with antibiotics, RSV neither decreased TMAO levels nor increased hepatic BA synthesis. Additionally, RSV-induced inhibition of TMAO-caused AS was also markedly abolished by antibiotics. In conclusion, RSV attenuated TMAO-induced AS by decreasing TMAO levels and increasing hepatic BA neosynthesis via gut microbiota remodeling, and the BA neosynthesis was partially mediated through the enterohepatic FXR-FGF15 axis. IMPORTANCE Recently, trimethylamine- N -oxide (TMAO) has been identified as a novel and independent risk factor for promoting atherosclerosis (AS) partially through inhibiting hepatic bile acid (BA) synthesis. The gut microbiota plays a key role in the pathophysiology of TMAO-induced AS. Resveratrol (RSV) is a natural phytoalexin with prebiotic benefits. A growing body of evidence supports the hypothesis that phenolic phytochemicals with poor bioavailability are possibly acting primarily through remodeling of the gut microbiota. The current study showed that RSV attenuated TMAO-induced AS by decreasing TMAO levels and increasing hepatic BA neosynthesis via gut microbiota remodeling. And RSV-induced hepatic BA neosynthesis was partially mediated through downregulating the enterohepatic farnesoid X receptor-fibroblast growth factor 15 axis. These results offer new insights into the mechanisms responsible for RSV’s anti-AS effects and indicate that the gut microbiota may become an interesting target for pharmacological or dietary interventions to decrease the risk of developing cardiovascular diseases.

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

Resveratrol (RSV) attenuated trimethylamine-N-oxide (TMAO)-induced atherosclerosis in mice by remodeling the gut microbiota, reducing microbial TMA production, and increasing hepatic bile acid synthesis. The bile acid effects were partially mediated by downregulation of the enterohepatic FXR-FGF15 axis.

Key findings

  • RSV attenuated TMAO-induced atherosclerosis in ApoE−/− mice; this protection was markedly abolished by antibiotics, indicating dependence on the gut microbiota.
  • RSV decreased serum TMA and TMAO levels by inhibiting gut microbial TMA production, while increasing hepatic FMO3 expression and FMO activity, so the TMAO reduction was not due to decreased liver FMO3.
  • RSV increased Lactobacillus and Bifidobacterium, increased bile salt hydrolase (BSH) activity, enhanced bile acid deconjugation and fecal bile acid excretion, decreased ileal bile acid content, repressed ileal FXR-FGF15, and increased hepatic CYP7A1 expression and bile acid neosynthesis. An FXR antagonist mimicked RSV effects on FGF15/CYP7A1, while an FXR agonist abolished them.
  • In choline-treated ApoE−/− mice, RSV increased Bacteroidetes abundance from 20.6% to 34.0% and decreased Firmicutes from 60.1% to 50.1%; RSV also increased Bacteroides, Lactobacillus, Bifidobacterium, and Akkermansia and decreased Prevotella, Ruminococcaceae_unclassified, and Biophila.

Methods

  • Female 8-week-old C57BL/6J and ApoE−/− mice were fed chow with or without RSV (0.4%), in the presence or absence of choline (1%) and/or antibiotics; some experiments used single doses of choline (400 mg/kg) or TMA (40 mg/kg). Treatment durations were 30 days for C57BL/6J studies and 4 months for ApoE−/− atherosclerosis studies, with n = 5 or 10 per group.
  • TMA/TMAO and bile acids were measured by LC/MS; gut microbiota was analyzed by 16S rRNA gene sequencing and qPCR; liver/ileal expression was assessed by qPCR and Western blotting; FMO and BSH activities were measured; atherosclerosis was assessed by ultrasound, Oil red O staining, and aortic cholesterol content.
  • Statistical analysis used Student’s t test or one-way ANOVA with Tukey-Kramer post hoc tests; correlations used Pearson’s tests with FDR adjustment for multiple comparisons.

Limitations

  • The paper states that the exact mechanisms by which circulating TMAO promotes atherosclerosis, whether TMAO acts through a specific receptor, and whether RSV reverses other TMAO-caused signaling changes remain largely unknown and need further elucidation.
  • The paper notes that RSV has poor bioavailability, and the exact mechanisms linking RSV, SIRT1-FXR signaling, FMO3 regulation, and bile acid transport need further clarification.
  • Evident from the text: findings are from female mouse models only, with no human clinical outcomes; antibiotic treatment is nonselective and may affect microbiota beyond TMA production, so microbiota-dependent conclusions are correlative and ablation-based rather than mechanism-specific.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1128/mbio.02210-15

Cites

49 of 49 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Figures

FIG 1 RSV inhibited TMAO synthesis in C57BL/6J mice. (A and B) Eight-week-old female C57BL/6J mice were administered choline (400 mg/kg of body weight, n = 10) (A) or TMA (40 mg/kg, n = 10) (B). Blood samples were collected at the indicated times. Serum TMA and TMAO levels were determined by LC/MS. (C and D) Eight-week-old female C57BL/6J mice were fed a chow diet with or without RSV (0.4%) in the presence or absence of choline (1%) or Abs. After 30 days, several mice of the chow-and-RSV-fed group were administered choline (400 mg/kg, n = 10) (C) or TMA (40 mg/kg, n = 10) (D). At 4 h after choline was given, or 1 h after TMA was given, the mice were euthanized and blood was collected. Serum TMA and TMAO levels were determined by LC/MS. (E and F) The other mice were also euthanized, and blood was collected. Serum TMA and TMAO levels (E) and RSV levels (F) were determined by LC/MS. Values are presented as means ± SD (n = 10). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus group cotreated with choline and RSV).
FIG 1 RSV inhibited TMAO synthesis in C57BL/6J mice. (A and B) Eight-week-old female C57BL/6J mice were administered choline (400 mg/kg of body weight, n = 10) (A) or TMA (40 mg/kg, n = 10) (B). Blood samples were collected at the indicated times. Serum TMA and TMAO levels were determined by LC/MS. (C and D) Eight-week-old female C57BL/6J mice were fed a chow diet with or without RSV (0.4%) in the presence or absence of choline (1%) or Abs. After 30 days, several mice of the chow-and-RSV-fed group were administered choline (400 mg/kg, n = 10) (C) or TMA (40 mg/kg, n = 10) (D). At 4 h after choline was given, or 1 h after TMA was given, the mice were euthanized and blood was collected. Serum TMA and TMAO levels were determined by LC/MS. (E and F) The other mice were also euthanized, and blood was collected. Serum TMA and TMAO levels (E) and RSV levels (F) were determined by LC/MS. Values are presented as means ± SD (n = 10). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus group cotreated with choline and RSV).CC BY · PubMed Central
FIG 2 RSV inhibited TMAO synthesis by decreasing TMA generation via remodeling microbiota in C57BL/6J mice. Eight-week-old female C57BL/6J mice (n = 5 per group) were fed chow with or without RSV (0.4%) for 30 days. ACTB, β-actin. (A) Western blotting detection of FMO3 in the liver. (B) Expression levels of FMO3 gene mRNAs were quantified using qPCR assays. (C) Liver FMO activity was assessed as described in Materials and Methods. (D) 16S rRNA gene sequencing analysis of cecal content at the phylum level. (E) Heat map of 16S rRNA gene sequencing analysis of cecal content at the genus level. The scale reflects the data as follows: red indicates high values whereas blue indicates low values for the percentages of reads that were classified at that rank. (F) Linear discriminant analysis (LDA) coupled with effect size measurements identifies the taxons most differentially abundant between the chow and RSV diets at the genus level. RSV-diet-enriched taxa are indicated with a positive LDA score (green), and taxa enriched in the normal chow diet have a negative score (red). Only taxa meeting an LDA significant threshold value of >2 are shown. (G) Correlation heat map demonstrating the association between the indicated microbiota taxonomic genera and TMA and TMAO concentrations (all reported as means ± SD in micromoles) from mice grouped by dietary status (chow and RSV). Red denotes a positive association, blue a negative association, and white no association. A single asterisk indicates a significant FDR-adjusted association at P values of ≤0.05, and a double asterisk indicates a significant FDR-adjusted association at P values of ≤0.01. (H) The production of TMA from choline by cecal content in vitro. (I) Cecal RSV content. (J) The same 8-week-old female C57BL/6J mice (n = 5 per group) were administered choline (400 mg/kg) at visit 1 (baseline), visit 2 (RSV treatment for 1 month), and visit 3 (washout for 1 month). Four hours after choline was given at each visit, blood samples were obtained from the tail veins. Serum TMA and TMAO levels were determined by LC/MS. Values are presented as means ± SD (n = 5). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); &, P < 0.01 (versus RSV-treated group).
FIG 2 RSV inhibited TMAO synthesis by decreasing TMA generation via remodeling microbiota in C57BL/6J mice. Eight-week-old female C57BL/6J mice (n = 5 per group) were fed chow with or without RSV (0.4%) for 30 days. ACTB, β-actin. (A) Western blotting detection of FMO3 in the liver. (B) Expression levels of FMO3 gene mRNAs were quantified using qPCR assays. (C) Liver FMO activity was assessed as described in Materials and Methods. (D) 16S rRNA gene sequencing analysis of cecal content at the phylum level. (E) Heat map of 16S rRNA gene sequencing analysis of cecal content at the genus level. The scale reflects the data as follows: red indicates high values whereas blue indicates low values for the percentages of reads that were classified at that rank. (F) Linear discriminant analysis (LDA) coupled with effect size measurements identifies the taxons most differentially abundant between the chow and RSV diets at the genus level. RSV-diet-enriched taxa are indicated with a positive LDA score (green), and taxa enriched in the normal chow diet have a negative score (red). Only taxa meeting an LDA significant threshold value of >2 are shown. (G) Correlation heat map demonstrating the association between the indicated microbiota taxonomic genera and TMA and TMAO concentrations (all reported as means ± SD in micromoles) from mice grouped by dietary status (chow and RSV). Red denotes a positive association, blue a negative association, and white no association. A single asterisk indicates a significant FDR-adjusted association at P values of ≤0.05, and a double asterisk indicates a significant FDR-adjusted association at P values of ≤0.01. (H) The production of TMA from choline by cecal content in vitro. (I) Cecal RSV content. (J) The same 8-week-old female C57BL/6J mice (n = 5 per group) were administered choline (400 mg/kg) at visit 1 (baseline), visit 2 (RSV treatment for 1 month), and visit 3 (washout for 1 month). Four hours after choline was given at each visit, blood samples were obtained from the tail veins. Serum TMA and TMAO levels were determined by LC/MS. Values are presented as means ± SD (n = 5). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); &, P < 0.01 (versus RSV-treated group).CC BY · PubMed Central
FIG 3 RSV enhanced BA deconjugation and fecal excretion in C57BL/6J mice. Eight-week-old female C57BL/6J mice (n = 10 per group) were fed chow with or without RSV (0.4%) in the presence or absence of Abs for 30 days. Fecal samples were collected. (A) Fecal BA excretion was measured by a total BA test (Wako). Fecal BA composition was determined by LC/MS as described in Materials and Methods. (B) CA/DCA ratio. (C) Conjugated/unconjugated (conj/unconj) BA ratio in fecal samples. (D) BSH activity. Values are expressed as means ± SD (n = 10). b, P < 0.01 (versus vehicle-treated control group); &, P < 0.01 (versus RSV-treated group).
FIG 3 RSV enhanced BA deconjugation and fecal excretion in C57BL/6J mice. Eight-week-old female C57BL/6J mice (n = 10 per group) were fed chow with or without RSV (0.4%) in the presence or absence of Abs for 30 days. Fecal samples were collected. (A) Fecal BA excretion was measured by a total BA test (Wako). Fecal BA composition was determined by LC/MS as described in Materials and Methods. (B) CA/DCA ratio. (C) Conjugated/unconjugated (conj/unconj) BA ratio in fecal samples. (D) BSH activity. Values are expressed as means ± SD (n = 10). b, P < 0.01 (versus vehicle-treated control group); &, P < 0.01 (versus RSV-treated group).CC BY · PubMed Central
FIG 4 RSV induced hepatic BA synthesis in C57BL/6J mice. Eight-week-old female C57BL/6J mice (n = 10 per group) were fed chow with or without RSV (0.4%) in the presence or absence of Abs for 30 days. (A) Liver cholesterol content. (B) The BA pool size was determined for the total BA content of gallbladder bile, liver, and the SI lumenal. (C to F) Total biliary BA content (C) and total BA content in SI lumenal (D), serum (E), and liver (F) were detected. (G) Gallbladder bile samples were subjected to LC/MS, and TCA/TβMCA ratios were calculated. (H) Relative expression levels of the indicated mRNAs in the liver were determined by qPCR. (I) Expression of CYP7A1 was analyzed by Western blotting. Values are expressed as means ± SD (n = 10). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); &, P < 0.01 (versus RSV-treated group).
FIG 4 RSV induced hepatic BA synthesis in C57BL/6J mice. Eight-week-old female C57BL/6J mice (n = 10 per group) were fed chow with or without RSV (0.4%) in the presence or absence of Abs for 30 days. (A) Liver cholesterol content. (B) The BA pool size was determined for the total BA content of gallbladder bile, liver, and the SI lumenal. (C to F) Total biliary BA content (C) and total BA content in SI lumenal (D), serum (E), and liver (F) were detected. (G) Gallbladder bile samples were subjected to LC/MS, and TCA/TβMCA ratios were calculated. (H) Relative expression levels of the indicated mRNAs in the liver were determined by qPCR. (I) Expression of CYP7A1 was analyzed by Western blotting. Values are expressed as means ± SD (n = 10). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); &, P < 0.01 (versus RSV-treated group).CC BY · PubMed Central
FIG 5 The enterohepatic FXR-FGF15 axis played a key role in RSV-induced BA synthesis. Eight-week-old female C57BL/6J mice (n = 10 per group) were fed chow with or without RSV (0.4%) for 30 days. Z-Gug (100 mg/kg body weight) or GW4064 (75 mg/kg body weight) was given 7 days prior to surgical procedures. (A) Relative expression levels of ileal FXR and FGF15 gene mRNAs. (B) Western blotting was used to detect ileal FXR and FGF15 expression. (C) Expression of the indicated proteins in ileal tissues. (D) Liver CYP7A1 expression was detected by Western blotting. (E) FGF15 expression in ileal tissues. (F) CYP7A1 expression in liver samples. Values are expressed as means ± SD (n = 10). b, P < 0.01 (versus vehicle-treated control group); &, P < 0.01 (versus RSV-treated group).
FIG 5 The enterohepatic FXR-FGF15 axis played a key role in RSV-induced BA synthesis. Eight-week-old female C57BL/6J mice (n = 10 per group) were fed chow with or without RSV (0.4%) for 30 days. Z-Gug (100 mg/kg body weight) or GW4064 (75 mg/kg body weight) was given 7 days prior to surgical procedures. (A) Relative expression levels of ileal FXR and FGF15 gene mRNAs. (B) Western blotting was used to detect ileal FXR and FGF15 expression. (C) Expression of the indicated proteins in ileal tissues. (D) Liver CYP7A1 expression was detected by Western blotting. (E) FGF15 expression in ileal tissues. (F) CYP7A1 expression in liver samples. Values are expressed as means ± SD (n = 10). b, P < 0.01 (versus vehicle-treated control group); &, P < 0.01 (versus RSV-treated group).CC BY · PubMed Central
FIG 6 RSV protected ApoE−/− mice from TMAO-induced AS. Eight-week-old female ApoE−/− mice (n = 10 per group) were given RSV (0.4%) with or without choline (1%) in the absence or presence of Abs for 4 months. The control group was fed with a chow diet. (A) Ultrasound B-mode images of the aortic sinus and quantification. Arrows indicate the regions of interest. RV, right ventricle; RA, right atrium; LA, left atrium. (B) Oil red O-stained aortic roots (counterstained with hematoxylin) and quantification. (C) Oil red O staining of whole aortas, including the aortic arch, thoracic, and abdominal regions, and their quantitation. (D) Total cholesterol content in the thoracic and abdominal aorta. Values are expressed as means ± SD (n = 10). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); c, P < 0.05; d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus group cotreated with RSV and choline).
FIG 6 RSV protected ApoE−/− mice from TMAO-induced AS. Eight-week-old female ApoE−/− mice (n = 10 per group) were given RSV (0.4%) with or without choline (1%) in the absence or presence of Abs for 4 months. The control group was fed with a chow diet. (A) Ultrasound B-mode images of the aortic sinus and quantification. Arrows indicate the regions of interest. RV, right ventricle; RA, right atrium; LA, left atrium. (B) Oil red O-stained aortic roots (counterstained with hematoxylin) and quantification. (C) Oil red O staining of whole aortas, including the aortic arch, thoracic, and abdominal regions, and their quantitation. (D) Total cholesterol content in the thoracic and abdominal aorta. Values are expressed as means ± SD (n = 10). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); c, P < 0.05; d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus group cotreated with RSV and choline).CC BY · PubMed Central
FIG 7 RSV reduced TMAO levels by decreasing TMA generation via remodeling microbiota in ApoE−/− mice. Eight-week-old female ApoE−/− mice (n = 5 per group) were fed chow, chow with RSV (0.4%), chow with choline (1%), or chow with choline (1%) plus RSV (0.4%) in the absence or presence of Abs for 4 months. (A) Serum TMA and TMAO levels were measured by LC/MS. (B) Western blotting detection of FMO3 expression in the liver. (C) Expression levels of FMO3 gene mRNAs were quantified using qPCR assays. (D) Liver FMO activity was assessed as described in Materials and Methods. (E) 16S rRNA gene sequencing analysis of cecal content at the phylum level. (F) Heat map of 16S rRNA gene sequencing analysis of cecal content at the genus level. The scale reflects the data as follows: red indicates high values whereas blue indicates low values for the spercentage of reads that were classified at that rank. (G) Linear discriminant analysis (LDA) coupled with effect size measurements identifies the most differentially abundant taxons at the genus level between the choline diet and the RSV-plus-choline diet. RSV-plus-choline-diet-enriched taxa are indicated with a positive LDA score (green), and taxa enriched in the choline diet have a negative score (red). Only taxa meeting an LDA significance threshold of >2 are shown. (H) Correlation heat map demonstrating the association between the indicated microbiota taxonomic genera and TMA and TMAO concentrations (all reported as means ± SD in micromoles) from mice grouped by dietary status (choline and RSV plus choline). Red denotes a positive association, blue a negative association, and white no association. A single asterisk indicates a significant FDR-adjusted association of P values of ≤0.05, and a double asterisk indicates a significant FDR-adjusted association of P values of ≤0.01. (I) Production of TMA from choline by cecal content in vitro. Values are expressed as means ± SD (n = 5). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); c, P < 0.05; d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus group cotreated with RSV and choline).
FIG 7 RSV reduced TMAO levels by decreasing TMA generation via remodeling microbiota in ApoE−/− mice. Eight-week-old female ApoE−/− mice (n = 5 per group) were fed chow, chow with RSV (0.4%), chow with choline (1%), or chow with choline (1%) plus RSV (0.4%) in the absence or presence of Abs for 4 months. (A) Serum TMA and TMAO levels were measured by LC/MS. (B) Western blotting detection of FMO3 expression in the liver. (C) Expression levels of FMO3 gene mRNAs were quantified using qPCR assays. (D) Liver FMO activity was assessed as described in Materials and Methods. (E) 16S rRNA gene sequencing analysis of cecal content at the phylum level. (F) Heat map of 16S rRNA gene sequencing analysis of cecal content at the genus level. The scale reflects the data as follows: red indicates high values whereas blue indicates low values for the spercentage of reads that were classified at that rank. (G) Linear discriminant analysis (LDA) coupled with effect size measurements identifies the most differentially abundant taxons at the genus level between the choline diet and the RSV-plus-choline diet. RSV-plus-choline-diet-enriched taxa are indicated with a positive LDA score (green), and taxa enriched in the choline diet have a negative score (red). Only taxa meeting an LDA significance threshold of >2 are shown. (H) Correlation heat map demonstrating the association between the indicated microbiota taxonomic genera and TMA and TMAO concentrations (all reported as means ± SD in micromoles) from mice grouped by dietary status (choline and RSV plus choline). Red denotes a positive association, blue a negative association, and white no association. A single asterisk indicates a significant FDR-adjusted association of P values of ≤0.05, and a double asterisk indicates a significant FDR-adjusted association of P values of ≤0.01. (I) Production of TMA from choline by cecal content in vitro. Values are expressed as means ± SD (n = 5). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); c, P < 0.05; d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus group cotreated with RSV and choline).CC BY · PubMed Central
FIG 8 RSV induced hepatic BA synthesis in choline-treated ApoE−/− mice. Eight-week-old female ApoE−/− mice (n = 10 per group) were fed chow, chow with RSV (0.4%), chow with choline (1%), or chow with choline (1%) plus RSV (0.4%) in the absence or presence of Abs for 4 months. (A) Serum TC levels were measured enzymatically using commercially available kits. (B) Liver cholesterol content. (C) The BA pool size was determined for the total BA content of gallbladder bile, liver, and the SI lumenal. BW, body weight. (D to F) Total biliary BA content (D) and total BA content in SI lumenal (E) and liver (F) were detected. (G) Relative expression levels of the indicated mRNAs in the liver were determined by qPCR. (H) Expression of CYP7A1 was analyzed by Western blotting. Values are expressed as means ± SD (n = 10). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); c, P < 0.05; d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus choline and RSV-cotreated group).
FIG 8 RSV induced hepatic BA synthesis in choline-treated ApoE−/− mice. Eight-week-old female ApoE−/− mice (n = 10 per group) were fed chow, chow with RSV (0.4%), chow with choline (1%), or chow with choline (1%) plus RSV (0.4%) in the absence or presence of Abs for 4 months. (A) Serum TC levels were measured enzymatically using commercially available kits. (B) Liver cholesterol content. (C) The BA pool size was determined for the total BA content of gallbladder bile, liver, and the SI lumenal. BW, body weight. (D to F) Total biliary BA content (D) and total BA content in SI lumenal (E) and liver (F) were detected. (G) Relative expression levels of the indicated mRNAs in the liver were determined by qPCR. (H) Expression of CYP7A1 was analyzed by Western blotting. Values are expressed as means ± SD (n = 10). a, P < 0.05; b, P < 0.01 (versus vehicle-treated control group); c, P < 0.05; d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus choline and RSV-cotreated group).CC BY · PubMed Central
FIG 9 RSV downregulated the enterohepatic FXR-FGF15 axis in choline-treated ApoE−/− mice. Eight-week-old female ApoE−/− mice (n = 10 per group) were fed chow, chow with RSV (0.4%), chow with choline (1%), or chow with choline (1%) plus RSV (0.4%) in the absence or presence of Abs for 4 months. (A) Expression of CYP7A1 was analyzed by Western blotting. (B and C) Relative expression levels of the indicated mRNAs in the liver (B) and ileal tissue (C) were determined by qPCR. (D) Western blotting was used to detect ileal FXR and FGF15 expression. (E and F) BA content in SI tissue (E) and fecal samples (F). b, P < 0.01 (versus vehicle-treated control group); d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus group cotreated with choline and RSV).
FIG 9 RSV downregulated the enterohepatic FXR-FGF15 axis in choline-treated ApoE−/− mice. Eight-week-old female ApoE−/− mice (n = 10 per group) were fed chow, chow with RSV (0.4%), chow with choline (1%), or chow with choline (1%) plus RSV (0.4%) in the absence or presence of Abs for 4 months. (A) Expression of CYP7A1 was analyzed by Western blotting. (B and C) Relative expression levels of the indicated mRNAs in the liver (B) and ileal tissue (C) were determined by qPCR. (D) Western blotting was used to detect ileal FXR and FGF15 expression. (E and F) BA content in SI tissue (E) and fecal samples (F). b, P < 0.01 (versus vehicle-treated control group); d, P < 0.01 (versus choline-treated group); #, P < 0.05 (versus group cotreated with choline and RSV).CC BY · PubMed Central
FIG 10 A model depicting use of RSV to attenuate AS by targeting gut microbiota. Dietary RSV can alter the composition of gut flora. On the one hand, this can result in reduced levels of gut microbial TMA production, subsequently leading to decreased TMAO synthesis in the liver and, ultimately, to inhibition of AS. On the other hand, RSV increases BSH activity by gut microbiota remodeling, which promotes the generation of unconjugated BAs from conjugated BAs and enhances fecal BA loss. RSV-induced fecal BA loss leads to a decrease in ileal BA content, thereby inhibiting the ileal FXR-FGF15 axis, and then increases the expression levels of CYP7A1 in the liver, subsequently inducing hepatic BA neosynthesis which contributes to cholesterol homeostasis, finally attenuating AS. Red indicates the effect of RSV, purple indicates downregulation by RSV, green indicates upregulation by RSV, and blue indicates the gut flora. LCA, lithocholic acid.
FIG 10 A model depicting use of RSV to attenuate AS by targeting gut microbiota. Dietary RSV can alter the composition of gut flora. On the one hand, this can result in reduced levels of gut microbial TMA production, subsequently leading to decreased TMAO synthesis in the liver and, ultimately, to inhibition of AS. On the other hand, RSV increases BSH activity by gut microbiota remodeling, which promotes the generation of unconjugated BAs from conjugated BAs and enhances fecal BA loss. RSV-induced fecal BA loss leads to a decrease in ileal BA content, thereby inhibiting the ileal FXR-FGF15 axis, and then increases the expression levels of CYP7A1 in the liver, subsequently inducing hepatic BA neosynthesis which contributes to cholesterol homeostasis, finally attenuating AS. Red indicates the effect of RSV, purple indicates downregulation by RSV, green indicates upregulation by RSV, and blue indicates the gut flora. LCA, lithocholic acid.CC BY · PubMed Central

Verification

Claims the paper makes

67 Own result · 41 Not checked yet · 37 Own interpretation · 2 Supported · 2 Partly supported · 1 Couldn’t confirm · 1 No basis given · accuracy not yet measured

Verify all 65 references — free account

Show all 84 claims

  • Probably not a substantial claim. 27
  • It may state two separate claims as one. 43
  • The quote only partly states this claim. 8
67 of 67
  • The paper relies on this without support.

    The mechanisms by which resveratrol exerts its anti-atherosclerosis effects remain unclear.

    “Resveratrol (RSV) is a natural phytoalexin with anti-AS effects; however, its mechanisms of action remain unclear.”
  • The paper relies on this without support.

    The gut microbiota is strongly associated with atherosclerosis.

    “The gut microbiota is found to be strongly associated with atherosclerosis (AS).”
  • The paper relies on this without support.

    Antibiotics have side effects and resistance potential that limit their utility.

    “However, the side effects and resistance potential of Abs limit their utility.”
  • The paper relies on this without support.

    Phenolic phytochemicals with poor bioavailability possibly act primarily through remodeling the gut microbiota.

    “However, a growing body of evidence supports the hypothesis that phenolic phytochemicals with poor bioavailability are possibly acting primarily through remodeling the gut microbiota.”
  • The paper relies on this without support.

    Antibiotic treatment suppresses the gut microbiota sufficiently to test its role in the effects of resveratrol.

    “Moreover, when the gut microbiota was suppressed by Abs, RSV-induced inhibition of TMAO-caused AS was significantly abolished (Fig. 6).”
  • The paper relies on this without support.

    Suppression of gut microbiota by antibiotics specifically suppresses gut microbiota without affecting other host pathways relevant to bile acid synthesis and atherosclerosis.

    “when the gut microbiota was suppressed by Abs”
    “in the absence or presence of Abs”
  • The paper relies on this without support.

    Z-Gug and GW4064 specifically modulate FXR without off-target effects on FGF15 and CYP7A1 expression.

    “When Z-Gug, a natural antagonist of FXR, was administered”
    “the presence of GW4064, a synthetic FXR agonist”
7 of 7

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.

Each contradiction between two passages listed here was confirmed by an independent check.

2 of 2

Citations

Cites

Show all 50 references

Research works printed but not linked: 2

  1. 10.1172/jci16309 · DOI · Paper page
  2. 10.1016/j.cmet.2013.01.003 · DOI · Paper page

Cited by

Sorted by citations

See all 754

Metadata

Authors
Chen, M.-L., Yi, L., Zhang, Y., Zhou, X., Ran, L., Yang, J., Zhu, J.-D., Zhang, Q.-Y., & Mi, M.-T.
Affiliations
  • Research Center for Nutrition and Food Safety, Institute of Military Preventive Medicine, Third Military Medical University, Chongqing, People’s Republic of China
  • Army Medical University (CN)
Venue
mBio
Published
2016-05-04
Volume
7
Issue
2
Pages
e02210-15
ISSN
  • 2150-7511 (ISSN-L)
  • 2161-2129
OpenAlex
W2324161694
Cited by
754
References
50
Citations by year
2026: 45 · 2025: 76 · 2024: 93 · 2023: 98 · 2022: 107 · 2021: 92 · 2020: 82 · 2019: 71 · 2018: 51 · 2017: 30 · 2016: 8
FWCI
27.88
Type
journal article
Language
en
Open access
Locations
  • mBio (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
National Natural Science Foundation of China (81470562)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gut microbiota and health
  • Health Sciences › Medicine › Geriatrics and Gerontology › Sirtuins and Resveratrol in Medicine
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Cholesterol and Lipid Metabolism
Keywords
resveratrol, trimethylamine N-oxide, atherosclerosis, gut microbiota, bile acid metabolism, bile salt hydrolase, farnesoid X receptor, FGF15, CYP7A1, hepatic bile acid synthesis, ApoE-deficient mice
MeSH terms
Gastrointestinal Microbiome, Resveratrol, Animals, Bacteria, Bile Acids and Salts, Cholesterol 7-alpha-Hydroxylase, Female, Humans, Liver, Methylamines, Mice, Inbred C57BL, Stilbenes, Atherosclerosis, Mice

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.

A language model drew the claims from the paper on 2026-10-07, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-10-07 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

Cited sources are checked with a language model’s help (gpt-6-sol).

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