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J Neurogastroenterol Motil. 2024;30(2):236-250.

The Effect of Clostridium butyricum on Gut Microbial Changes and Functional Profiles of Metabolism in High-fat Diet-fed Rats Depending on Age and Sex.

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제1저자 최수인 · 교신저자 김나영

소속 (4)
  1. Department of Internal Medicine, Seoul National University Bundang Hospital, Seongnam, Gyeonggi-do, Korea.
  2. Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Korea.
  3. Department of Medical Device Development, Seoul National University College of Medicine, Seoul, Korea.
  4. Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, Korea.

Abstract

BACKGROUND/AIMS A high-fat diet (HFD) causes dysbiosis and promotes inflammatory responses in the colon. This study aims to evaluate the effects of Clostridium butyricum on HFD-induced gut microbial changes in rats.

METHODS Six-week-old Fischer-344 rats with both sexes were given a control or HFD during 8 weeks, and 1-to-100-fold diluted Clostridium butyricum were administered by gavage. Fecal microbiota analyses were conducted using 16S ribosomal RNA metagenomic sequencing and predictive functional profiling of microbial communities in metabolism.

RESULTS A significant increase in Ruminococcaceae and Lachnospiraceae, which are butyric acid-producing bacterial families, was observed in the probiotics groups depending on sex. In contrast, Akkermansia muciniphila, which increased through a HFD regardless of sex, and decreased in the probiotics groups. A. muciniphila positively correlated with Claudin-1 expression in males (P < 0.001) and negatively correlated with the expression of Claudin-2 (P = 0.042), IL-1β (P = 0.037), and IL-6 (P = 0.044) in females. In terms of functional analyses, a HFD decreased the relative abundances of M00131 (carbohydrate metabolism module), M00579, and M00608 (energy metabolism), and increased those of M00307 (carbohydrate metabolism), regardless of sex. However, these changes recovered especially in male C. butyricum groups. Furthermore, M00131, M00579, and M00608 showed a positive correlation and M00307 showed a negative correlation with the relative abundance of A. muciniphila (P < 0.001).

CONCLUSION The beneficial effects of C. butyricum on HFD-induced gut dysbiosis in young male rats originate from the functional profiles of carbohydrate and energy metabolism.

Keywords

Clostridium butyricumGastrointestinal microbiomeHigh-fat dietMetabolismProbiotics

Figures

Figure 1
Figure 1 The characteristics of the stool microbiome. (A, B) The composition of each group at the phylum level. Bacteroidetes were commonly decreased by feeding rats with a high-fat diet (HFD). (C, D) The species richness was estimated by the Chao1 index. The Chao1 index decreased in the HFD group compared with the control group, regardless of sex, which was recovered by probiotic administration. (E, F) The Shannon index decreased in the HFD group compared with the control group, regardless of sex. M, male; F, female; CON, control diet; Bio, HFD with Biovita administration; LCB, HFD with a low concentration of Clostridium butyricum; MCB, HFD with a medium concentration of C. butyricum; and HCB, HFD with a high concentration of C. butyricum.
Figure 2
Figure 2 The compositional change of high-fat diet (HFD) and probiotic administration at the (A-D) family and (E, F) species levels. (A, B) Ruminococcaceae decreased in the HFD group regardless of sex, and especially increased in the female group administered a high concentration of Clostridium butyricum (P = 0.036) compared with the HFD group. (C, D) Lachnospiraceae family increased in female HFD group. However, C. butyricum administration increased Lachnospiraceae in the male group (low concentration, P = 0.021; high concentration, P = 0.027) compared with the HFD group. (E, F) Compared with the HFD group, Akkermansia muciniphila species increased in the HFD group regardless of sex, and decreased in the male group administered low and high concentrations of C. butyricum (low, P = 0.003; high, P = 0.027) and the female group administered a high concentration of C. butyricum (P = 0.030).
Figure 3
Figure 3 Correlation between Akkermansia muciniphila and the gene expression of colonic epithelial tight junction protein and inflammatory cytokines. A. muciniphila positively correlated with claudin-1 (cldn1) gene expression in males rats (A, P < 0.001), and negatively correlated with claudin-2 (cldn2) gene expression in females rats (B, P = 0.042). Conversely, in females, the relative abundance of A. muciniphila negatively correlated with inflammatory cytokines, IL-1β (C, P = 0.037), and IL-6 (D, P = 0.044). The statistical significance was determined by Spearman’s correlation. CON, control diet; HFD, high-fat diet; Bio, HFD with Biovita administration; LCB, HFD with a low concentration of Clostridium butyricum; MCB, HFD with a medium concentration of C. butyricum; HCB, HFD with a high concentration of C. butyricum.
Figure 4
Figure 4 Predictive functional profiling of microbial communities through the Kyoto Encyclopedia of Genes and Genomes (KEGG) database regarding carbohydrate metabolism based on KEGG modules. (A) The modules that showed significant difference between groups are represented as a heatmap, depending on the sex. The significantly different modules are shown in the scatter graph. (B) M00131, inositol phosphate metabolism, Ins(1,3,4,5)P4 → Ins(1,3,4)P3 → myo-inositol; (C) M00307, Pyruvate oxidation, pyruvate → acetyl-CoA. Akkermansia muciniphila showed negative correlation with M00131 (D, P < 0.001) and positive correlation with M00307 (E, P < 0.001). CON, control diet; HFD, high-fat diet; Bio, HFD with Biovita administration; LCB, HFD with a low concentration of Clostridium butyricum; MCB, HFD with a medium concentration of C. butyricum; HCB, HFD with a high concentration of C. butyricum.
Figure 5
Figure 5 Predictive functional profiling of microbial communities through the Kyoto Encyclopedia of Genes and Genomes (KEGG) database regarding energy metabolism based on KEGG modules. (A) The modules that showed significant differences between groups are represented as a heatmap, depending on the sex. The significantly different modules are shown in the scatter graph. (B) M00579, phosphate acetyltransferase-acetate kinase pathway, acetyl-CoA → acetate; (C) M00608, 2-oxocarboxylic acid chain extension, 2-oxoglutarate → 2-oxoadipate → 2-oxopimelate → 2-oxosuberate. Akkermansia muciniphila showed negative correlations with M00131 (D, P < 0.001) and M00307 (E, P < 0.001). CON, control diet; HFD, high-fat diet; Bio, HFD with Biovita administration; LCB, HFD with a low concentration of Clostridium butyricum; MCB, HFD with a medium concentration of C. butyricum; HCB, HFD with a high concentration of C. butyricum.
Figure 6
Figure 6 Graphical summary of the change of gut microbiota by a high-fat diet and Clostridium butyricum used as a probiotic. SCFAs, short-chain fatty acids.

그림 출처: PubMed Central 공개 논문 (PMC10999835)

Conflict of interest statement

Conflicts of interest: None.

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Journal of neurogastroenterology and motility · ISSN 2093-0879
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