Gut microbiome mediates the protective effects of exercise after myocardial infarction

Figures

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  Fig. 1. Running training protects against cardiac dysfunction after MI. A The schedule of running training after MI. B Running increased ejection fractions (EF) and fractional shortening (FS) post-MI (n = 10:12:17:12). C Running increased endurance capacity post-MI (n = 10:12:17:12). D, E Running decreased cardiac ANP and BNP mRNA expression post-MI (n = 6:6:6:6). F Running decreased cardiac fibrosis post-MI (n = 10:12:12:11). G Running decreased cardiac cross sectional area post-MI by H&E (n = 7:8:8:8). H Running decreased cardiac cross sectional area post-MI by WGA (n = 9:9:9:9). I Running decreased cardiac Bax/Bcl2, cleaved Caspase3/Caspase3 and Collagen1 post-MI (n = 6:6:6:6). Scale bar: 50 μm in F and 25 μm in G and H. Data were represented as mean ± SD. Significant differences were assessed by two-way ANOVA followed by Bonferroni’s multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001

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  Fig. 2. Running training changes gut microbial richness and community structure in mice after MI. A Pan analysis for four groups based on genus level (n = 7:18:12:12). B Fecal bacterial community at the phylum level among Sham + control, MI + control, Sham + Run, and MI + Run. C PCoA analysis based on the relative abundance of genus between Sham + control and MI + control groups. D PCoA analysis based on the relative abundance of genus between MI + control and MI + Run groups. E A total of 49 samples were clustered into enterotype 1 (green), enterotype 2 (red), and enterotype 3 (blue) at the genus level. F The percentage of Sham + control, MI + control, Sham + Run, and MI + Run samples distributed in three enterotypes. G Five changed genera among Sham + control, MI + control, Sham + Run, and MI + Run. Significant differences were assessed by ANOSIM analysis in C, D; abund jaccard analysis in E; Wilcoxon rank-sum with FDR in F. *p < 0.05, **p < 0.01, ***p < 0.001

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  Fig. 3. Gut microbiota pre-depletion by antibiotics abolishes the protection of running in MI. A Antibiotics (ABX and 1/4 ABX) decreased EF and FS in MI + Run mice (n = 8:10:10). B Antibiotics decreased endurance capacity in MI + Run mice (n = 7:7:7). C, D Antibiotics increased cardiac ANP and BNP mRNA expression in MI + Run mice (n = 6:6:6). E Antibiotics increased cardiac fibrosis in MI + Run mice (n = 7:6:8). F Antibiotics increased cardiac cross sectional area in MI + Run mice by H&E (n = 6:7:5). G Antibiotics increased cardiac cross sectional area in MI + Run mice by WGA (n = 8:8:8). H Antibiotics increased Bax/Bcl2, cleaved Caspase3/Caspase3 and Collagen1 in MI + Run mice (n = 6:6:6). Scale bar: 50 μm in E and 25 μm in F and G. Data were represented as mean ± SD. Significant differences were assessed by one-way ANOVA followed by Bonferroni's multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001

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  Fig. 4. FMT recovers the protection of running in MI. A The schedule of FMT after ABX treated. B FMT from MI + Run increased EF and FS compared to FMT from MI + Control post-MI (n = 14:14). C FMT from MI + Run increased endurance capacity compared to FMT from MI + Control post-MI (n = 11:11). D, E FMT from MI + Run decreased cardiac ANP and BNP mRNA expression compared to FMT from MI + control post-MI (n = 6:6). F FMT from MI + Run decreased cardiac fibrosis compared to FMT from MI + control post-MI (n = 9:9). G FMT from MI + Run decreased cardiac cross sectional area compared to FMT from MI + control post-MI by H&E (n = 8:8). H FMT from MI + Run decreased cardiac cross sectional area compared to FMT from MI + control post-MI by WGA (n = 12:14). I FMT from MI + Run decreased Bax/Bcl2, cleaved Caspase3/Caspase3 and collagen1 compared to FMT from MI + Control post-MI (n = 6:6). Scale bar: 50 μm in F and 25 μm in G and H. Data were represented as mean ± SD. Significant differences were assessed by two-tailed student t test. **p < 0.01, ***p < 0.001

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  Fig. 5. 3-HPA and 4-HPA are identified to be involved in the protection of running in MI. OPLS-DA Model Discrimination based on metabolic profiles in fecal samples. A The relative abundance of each metabolite classes in different groups (n = 10:10:10:10). B The abundance of phenols in different groups. C Volcano plot of univariate statistics across Sham + control group and MI + control group. D Volcano plot of univariate statistics across Sham + control group and Sham + Run group. E Pathway analysis bubble plot by mmu set in MI + Control vs MI + Run. F Network for statistically significant changed pathways in MI + control vs. MI + Run. G The relationship between metabolites and the top 50 genera in four groups was estimated by Spearman’s correlation analysis. Those with low correlated (|r|< 0.1) were not shown. Genera and cardiac function index were distinguished as positive (red) and negative (blue) correlation. *p < 0.05; **p < 0.01; ***p < 0.001

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  Fig. 6. 3-HPA and 4-HBA protect against cardiac dysfunction after MI. A 3-HPA and 4-HBA increased EF and FS post-MI (n = 10:10:9:11:9:11:11). B 3-HPA and 4-HBA increased endurance capacity post-MI (n = 10:11:10:10:11:11:11). C, D 3-HPA and 4-HBA decreased cardiac ANP and BNP mRNA expression post-MI (n = 6:6:6:6). E 3-HPA and 4-HBA decreased cardiac fibrosis post-MI (n = 7:7:7:7). F 3-HPA and 4-HBA decreased cardiac cross sectional area post-MI by H&E (n = 8:8:8:8). G 3-HPA and 4-HBA decreased cardiac cross sectional area post-MI by WGA (n = 9:9:9:9). H 3-HPA and 4-HBA decreased Bax/Bcl2, cleaved Caspase3/Caspase3 and Collagen1 post-MI (n = 6:6:6:6). Scale bar: 50 μm in E and 25 μm in F and G. Data were represented as mean ± SD. Significant differences were assessed by one-way ANOVA followed by Bonferroni's multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001

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  Fig. 7. 3-HPA and 4-HBA mediate cardiac protection through activating NRF2. A, B 3-HPA (100 μM, 24 h) reduced the percentage of TUNEL staining positive cardiomyocytes (n = 6:6:6:6), Bax/Bcl2, cleaved Caspase3/Caspase3, NRF2 (n = 3:3:3:3) in NRCMs under oxygen glucose deprivation/reperfusion (OGD/R). C, D 4-HBA (100 μM, 24 h) decreased the percentage of TUNEL staining positive cardiomyocytes (n = 6:6:6:6), Bax/Bcl2, cleaved Caspase3/Caspase3, NRF2 (n = 3:3:3:3) in NRCMs under OGD/R. E, F Inhibition of NRF2 increased the percentage of TUNEL staining positive cardiomyocytes decreased by 3-HPA and 4-HBA in NRCMs under OGD/R (n = 6:6:6:6). Scale bar: 100 μm. Data were represented as mean ± SD. Significant differences were assessed by two-way ANOVA followed by Bonferroni's multiple comparisons test. *p < 0.05, **p < 0.01, ***p < 0.001 versus respective control