Parabacteroides distasonis ameliorates insulin resistance via activation of intestinal GPR109a - PubMed (original) (raw)
. 2023 Nov 25;14(1):7740.
doi: 10.1038/s41467-023-43622-3.
Yonggan Sun # 1 2 3, Qixing Nie # 1 2 3, Shanshan Zhang 1 2 3, Huijun He 1 2 3, Sheng Zuo 1 2 3, Chunhua Chen 1 2 3, Jingrui Yang 1 2 3, Haihong Chen 1 2 3, Jielun Hu 1 2 3, Song Li 1 2 3, Baojie Zhang 1, Zhitian Zheng 1, Shijie Pan 1, Ping Huang 4, Lu Lian 4, Shaoping Nie 5 6 7
Affiliations
- PMID: 38007572
- PMCID: PMC10676405
- DOI: 10.1038/s41467-023-43622-3
Parabacteroides distasonis ameliorates insulin resistance via activation of intestinal GPR109a
Yonggan Sun et al. Nat Commun. 2023.
Abstract
Gut microbiota plays a key role in insulin resistance (IR). Here we perform a case-control study of Chinese adults (ChiCTR2200065715) and identify that Parabacteroides distasonis is inversely correlated with IR. Treatment with P. distasonis improves IR, strengthens intestinal integrity, and reduces systemic inflammation in mice. We further demonstrate that P. distasonis-derived nicotinic acid (NA) is a vital bioactive molecule that fortifies intestinal barrier function via activating intestinal G-protein-coupled receptor 109a (GPR109a), leading to ameliorating IR. We also conduct a bioactive dietary fiber screening to induce P. distasonis growth. Dendrobium officinale polysaccharide (DOP) shows favorable growth-promoting effects on P. distasonis and protects against IR in mice simultaneously. Finally, the reduced P. distasonis and NA levels were also validated in another human type 2 diabetes mellitus cohort. These findings reveal the unique mechanisms of P. distasonis on IR and provide viable strategies for the treatment and prevention of IR by bioactive dietary fiber.
© 2023. The Author(s).
Conflict of interest statement
The authors declare no competing interests.
Figures
Fig. 1. Decreased abundance of P. distasonis is associated with IR.
The 16S rRNA gene amplicon sequencing datasets displayed the change of fecal microbiota in T2DM (n = 30) and HC (n = 30) groups. a–c α-diversity indicated by the Shannon, ACE, and Chao1 index. The box plots showing the minima, maxima, center, bounds of box and whiskers and percentile. d Principal coordinate analysis (PCoA) of all samples by weighted UniFrac distance (ANOSIM test). e Taxonomic cladogram generated from LEfSe analysis of 16S rRNA gene sequences. Each circle’s size is proportional to the taxon’s abundance. f LDA score representing the taxonomic data with significant differences between T2DM and HC groups. Only LDA scores >3 are shown. Blue indicates enriched taxa in the T2DM group. Red indicates enriched taxa in the HC group. g Spearman correlations (two-tailed Spearman’s rank test) between the genus-level abundance and IR phenotypes. The color represents positive (red) or negative (blue) correlations and FDRs are denoted: *FDR < 0.05; **FDR < 0.01. h The relative abundance of fecal P. distasonis, P. merdae, P. johnsonii, and P. gordonii was determined by qPCR. Data are presented as the mean ± SEM. n = 30 per group. i The relative abundance of fecal F. prausnitzii was determined by qPCR. Data are presented as the mean ± SEM. n = 30 per group. j Spearman correlations (two-tailed Spearman’s rank test) between the species-level abundance and IR phenotypes. The color represents positive (red) or negative (blue) correlations and FDRs are denoted: *, FDR < 0.05; **, FDR < 0.01. T2DM (type 2 diabetes mellitus); HC (health control). Statistical analysis was performed using two-tailed Mann–Whitney test for a–c, h, and i. Source data are provided as a Source Data file.
Fig. 2. P. distasonis is selectively enriched by DOP.
a Schematic of P. distasonis enrichment by DOP in vitro. Briefly, we incubated these three dietary fibers (DOP, or inulin or β-glucan) independently with fecal bacteria from T2DM patients for 48 h. The majority of the fermented broth underwent centrifugation, while the resulting precipitation was used for 16S rRNA gene sequencing. Simultaneously, the supernatant was employed for the sugar content determination. Another portion of the fermented broth was coated in BHI agar medium, resulting in the isolation of 43 strains of P. distasonis. Finally, the 43 strains of P. distasonis were cultured in BHI medium (DOP as the sole carbon) for 24 h, and the OD600 was measured. The strain with the highest △OD600 was selected for further experiments. The tube in the image is referenced from Figure 1c of Ziesack et al (10.1128/msystems.00352-19) with appropriate color modifications. b Relative abundance of bacteria at the genus level in different groups. c Principal coordinate analysis (PCoA) of DOP and Vehicle group by weighted UniFrac distance (ANOSIM test). d Taxonomic cladogram generated from LEfSe analysis (DOP and Vehicle group) by 16S rRNA gene amplicon sequencing. Each circle’s size is proportional to the taxon’s abundance. e LDA score representing the taxonomic data with significant differences between DOP and Vehicle groups. Only LDA scores >2 are shown. Green indicates enriched taxa in the Vehicle group. Red indicates enriched taxa in the DOP group. f The relative abundance of P. distasonis in DOP and Vehicle group was determined by qPCR. n = 30 per group. g The relative abundance of P. merdae, P. johnsonii, P. goldsteinii, and P. gordonii in DOP and Vehicle groups were determined by qPCR. n = 30 per group. h–i Mice were treated with 400 mg/kg DOP by daily gavage for 7 days. h Experimental scheme for i. n = 6 mice per group. i The change of relative abundance of fecal P. distasonis by DOP treatment. DOP (Dendrobium officinale polysaccharide); BG (β-glucan); IN (inulin). Data are presented as the mean ± SEM. Statistical analysis was performed using two-tailed Mann–Whitney test for f and g, two-tailed paired _t_-test for i. Source data are provided as a Source Data file.
Fig. 3. DOP enrich P. distasonis and ameliorates HFD-induced IR.
After an 8-week HFD treatment, mice were given PBS (HFD group) or DOP (DOP group) for 5 weeks, and the control group (Ctrl) was fed with a chow diet and given an equivalent volume of PBS. a Experimental scheme for (b–p). b–l n = 8 mice per group, m–p n = 6 mice per group. b Body weight curve. c Body weight change (%). OGTT (d) and AUC (e). ITT (f) and AUC (g). h Fasting insulin level. i HOMA-IR index. j Serum TNF-α level. k Serum IL-1β level. l Serum LPS level. m Relative mRNA expression of genes related to intestinal permeability. n Claudin-1, Muc-2, Occludin, and ZO-1 immunohistologic staining of colonic sections (top), and quantified positive area (bottom). Scale bars, 50 μm. o LDA score represents the taxonomic data with significant differences between DOP and HFD group. Only LDA scores >3 are shown. Green indicates enriched taxa in the HFD group. Red indicates enriched taxa in the DOP group. p The relative abundance of P. distasonis in different groups assessed by qPCR. q Spearman correlations (two-tailed Spearman’s rank test) between the fecal P. distasonis abundance and IR phenotypes. DOP (Dendrobium officinale polysaccharide); HFD (High-fat diet). Data are presented as the mean ± SEM. Statistical analysis was performed using One-way ANOVA with Tukey’s post hoc test for b, 0, 15, 60, and 90 min of d, e, 60 and 90 min of f, h, i, j, m, Claudin-1, Occludin, and Zo-1 of n, One-way ANOVA with Dunnett’s T3 post hoc test for c, 30 min of d, 30 min of f, g, k, l, and p, Kruskal–Wallis test for 0 and 15 min of f, and Muc-2 of n. *P < 0.05; **P < 0.01. Source data are provided as a Source Data file.
Fig. 4. P. distasonis ameliorates HFD-induced IR.
After an 8-week HFD treatment, mice were given PBS (Vehicle group), P. distasonis NSP007 (LPD group), or heat-killed P. distasonis NSP007 (KPD group) for 5 weeks. a Experimental scheme for b–o. b–m n = 8 mice per group, n–o n = 6 mice per group. b The relative abundance of P. distasonis in different groups assessed by qPCR. c Body weight curve. d Body weight change (%). OGTT (e) and AUC (f). ITT (g) and AUC (h). i Fasting insulin level. j HOMA-IR index. k Serum TNF-α level. l Serum IL-1β level. m Serum LPS level. n Relative mRNA expression of genes related to intestinal permeability. o Claudin-1, Muc-2, Occludin, and ZO-1 immunohistologic staining of colonic sections (left), and quantified positive area (right). Scale bars, 50 μm. Data are presented as the mean ± SEM. Statistical analysis was performed using One-way ANOVA with Tukey’s post hoc test for b–d, 0 and 30 min of e, 0 and 30 min of g, j, k, m, Claudin1, Occludin, and Zo1 of n, and o, One-way ANOVA with Dunnett’s T3 post hoc test for 60 and 90 min of e, f, i, and Muc2 of n. Kruskal–Wallis test for 15 min of e, 15, 60, and 90 min of g, h, and l. *P < 0.05; **P < 0.01. Source data are provided as a Source Data file.
Fig. 5. Nicotinic acid is a bioactive metabolite of P. distasonis.
a–e Non-targeted metabolomic analysis of the BHI medium inoculated with P. distasonis or not. n = 3 biologically independent samples per group. a OPLS-DA analysis of metabolic profile in the supernatant of P. distasonis and BHI medium. b The enriched pathways calculated from mummichog algorithm based on the nontargeted metabolomics analysis of samples from P. distasonis incubation (hypergeometric test). c Enrichment analysis of the top 25 enrichment metabolic pathways (hypergeometric test). d Heat map of metabolites involved in the significantly changed metabolic pathways. e The MS/MS spectra of NA standard and NA detected in _P. distasonis_-incubated samples. f Extracted ion chromatograms of NA from cultured P. distasonis compared to the Vehicle. g The intensity of NA in the culture supernatant of P. distasonis at different time points. n = 3 biologically independent samples per group. h The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the encoded genes of P. distasonis NSP007. i–k Changes in the P. distasonis abundance and NA levels in the feces of mice before and after gavage with P. distasonis NSP007. Mice were treated with P. distasonis (2 × 108 CFU/mice/d) for 7 days. n = 12 mice per group. i Experimental scheme for j–k. j The relative abundance of P. distasonis. k The changes of fecal NA level. l Colonic NA level in Vehicle, LPD, and KPD group of mice. n = 8 mice per group. m Colonic NA level in HFD and DOP group of mice. n = 8 mice per group. n Spearman correlations (two-tailed Spearman’s rank test) between the colonic NA level and abundance of fecal P. distasonis in mice (Vehicle, LPD, KPD, HFD, and DOP group). o Fecal NA level in HC and T2DM group of humans. p Spearman correlations (two-tailed Spearman’s rank test) between the colonic NA level and fecal P. distasonis abundance (HC and T2DM group). q, r Level of fecal NA in mice after antibiotic cocktails (Abx) or fecal microbiota transplantation (FMT) treatment. n = 8 mice per group. q Experimental scheme for r. r Fecal NA level. DOP (Dendrobium officinale polysaccharide); HFD (High-fat diet); LPD (P. distasonis NSP007); KPD (heat-killed P. distasonis NSP007); T2DM (type 2 diabetes mellitus); HC (health control). Data are presented as the mean ± SEM. Statistical analysis was performed using One-way ANOVA with Tukey’s post hoc test for l, two-tailed paired _t_-test for j and k, and two-tailed unpaired _t_-test for m and o. Source data are provided as a Source Data file.
Fig. 6. NA improves intestinal integrity and IR.
a, b Caco-2 cells were treated with serum-free medium (Control) or medium added with NA for 12 h. n = 3 biologically independent samples per group. a Gpr109a expression in Caco-2 cells was examined using qPCR. b Claudin1, Muc2, Occludin, and Zo1 expression in Caco-2 cells was examined using qPCR. c Gpr109a, Claudin1, Muc2, Occludin, and Zo1 expression in Caco-2 cells was examined using qPCR. Caco-2 cells were pre-treated with serum-free medium (Control) or MPN for 1 h and subsequently stimulated with NA for another 12 h. n = 3 biologically independent samples per group. d Gpr109a expression in colonic tissues of mice in the HFD and DOP group. n = 6 mice per group. e Gpr109a expression in colonic tissues of mice in the HFD-FMT and DOP-FMT groups. n = 6 mice per group. f Gpr109a expression in colonic tissues of mice in the Vehicle, LPD, and KPD groups. n = 6 mice per group. g–t After an 8-week HFD treatment, mice were treated with PBS (Vehicle group), NA (NA group), MPN (Mepenzolate bromide, MPN group), or a combination of NA and MPN (NA + MPN group) three times a week for 5 weeks. h–r n = 8 mice per group, s–t n = 6 mice per group. g Experimental scheme for g–t. h Body weight curve. i Body weight change (%). j OGTT and k AUC. l ITT and m AUC. n Fasting insulin level. o HOMA-IR index. p Serum TNF-α level. q Serum IL-1β level. r Serum LPS level. s Relative mRNA expression of genes related to intestinal permeability. t Relative mRNA expression of Gpr109a in different groups. DOP (Dendrobium officinale polysaccharide); HFD (High-fat diet); LPD (P. distasonis NSP007); KPD (Heat-killed P. distasonis NSP007). Data are presented as the mean ± SEM. Statistical analysis was performed using One-way ANOVA with Tukey’s post hoc test for a–c, f, h–k, 15 min of l–o, q–t, One-way ANOVA with Dunnett’s T3 post hoc test for p, two-tailed unpaired _t_-test for d, two-tailed Kruskal–Wallis test for e and 30 min of l. *P < 0.05; **P < 0.01. Source data are provided as a Source Data file.
Fig. 7. P. distasonis abundance and fecal NA levels are negatively correlated with IR in human.
a The relative abundance of P. distasonis in HC (n = 30) and T2DM (n = 30) groups. b The level of fecal NA in HC (n = 30) and T2DM (n = 30) groups. c Spearman correlations (two-tailed Spearman’s rank test) between the fecal NA level and P. distasonis abundance in the validation cohort. Spearman correlations (two-tailed Spearman’s rank test) between the P. distasonis abundance and (d) FBG (fasting blood glucose) level, (e) insulin level, and (f) HOMA-IR (homeostasis model assessment measure of insulin resistance) index in the validation cohort. Spearman correlations (two-tailed Spearman’s rank test) between the fecal NA level and (g) FBG, (h) insulin level, and (i) HOMA-IR index in the validation cohort. Spearman correlations (two-tailed Spearman’s rank test) between the abundances of Parabacteroides and (j) BMI (body mass index), (k) FPG (Fasting plasma glucose) level, and (l) SSPG (steady-state plasma glucose) level. The data is from the Human Microbiome Project (
). T2DM (type 2 diabetes mellitus); HC (health control). Data are presented as the mean ± SEM. Statistical analysis was performed using two-tailed Mann–Whitney test for a and unpaired _t_-test for b. Source data are provided as a Source Data file.
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