Fig 1.
Loss of the gut microbiota aggravated helminth-induced cardiac fibrosis.
(A) Experimental scheme of Ts-infected mice received antibiotics (ABX) treatment. (B) Whole heart images of the controls (Con) (n = 6), Ts-infected mice (Ts) (n = 6) and Ts-infected mice with antibiotics (Ts+ABX) (n = 6). Representative images are shown. (C) Heart mass-to-body weight ratio (HM/BW). (D and E) Ejection fraction (EF) and left ventricular weight (LVM) obtained by cardiac ultrasound. (F, G and H) qPCR analysis of Collagen-1 (Col1), α-SMA and TGF-β expression in the heart tissue of mice. (I and J) Masson staining and Col1 immunohistochemistry results of heart tissues. Magnification, 200×. Scale bars, 100 μm. Representative images are shown. Data are shown as individual data points and mean ± SD. Data were compared by one-way ANOVA followed by Tukey multiple comparison tests. ns, not significant; *, p < 0.05; **, p < 0.01, ***, p <0.001, ****, p <0.0001.
Fig 2.
Characterization of the gut microbiota dysbiosis in Ts-infected mice with antibiotics (ABX) treatment using 16s rRNA sequencing.
(A) α diversity (Shannon index and Simpson index) in the microbiota from the controls (Con), Ts-infected mice and Ts-infected mice with antibiotics treatment (TsABX). (B) Principal component analysis (PCA) and principal coordinates analysis (PCoA) plot (β diversity). (C) Phylum and genus-level median relative abundances. (D) Linear discriminant analysis (LDA) effect size showing the most significantly differentially abundant taxa enriched in the microbiota. (E) Comparison proportion of genus levels of Akkermansia based on 16S rRNA gene sequencing.
Fig 3.
Restoration of gut microbiome alleviated helminth-induced cardiac fibrosis.
(A) Experimental scheme of Ts-infected mice received healthy gut microbiota. (B) Whole heart images of Ts-infected mice (Ts) (n = 6) and Ts-infected mice received healthy fecal microbiota transplantation (FMT) (Ts+FMT-H) (n = 6). Representative images are shown. (C) Heart mass-to-body weight ratio (HM/BW). (D and E) Ejection fraction (EF) and left ventricular weight (LVM) obtained by cardiac ultrasound. (F and G) qPCR analysis of Collagen-1 (Col1) and α-SMA expression in the heart tissue of mice. (H and I) Masson staining and Col1 immunohistochemistry results of heart tissues. Magnification, 200×. Scale bars, 100 μm. Representative images are shown. Data are shown as individual data points and mean ± SD. Statistical significance is calculated using paired student t-test. ns, not significant; *, p < 0.05; **, p < 0.01, ***, p <0.001, ****, p <0.0001.
Fig 4.
Characterization of the gut microbiota in Ts-infected mice using metagenomic sequencing.
(A) Chao1 index, Shannon index and Simpson index was analyzed by metagenomic. Data are shown as individual data points and mean ± SD. Difference was tested with Wilcoxon signed-rank test. (B) Principal component analysis (PCA) and principal coordinates analysis (PCoA) plot (β diversity) in the microbiota from the controls (Con) (n = 6) and Ts-infected mice (n = 6). (C) Phylum-level and genus-level median relative abundances of Con and Ts group. (D) Krona plot of identified bacteria species of Con and Ts group. (E) Comparison proportion of genus and species levels of Akkermansia muciniphila based on metagenomics.
Fig 5.
Both alive and pasteurized Akkermansia muciniphila had beneficial effect on helminth-induced cardiac fibrosis.
(A) Experimental scheme of alive or pasteurized A. muciniphila administration (8 × 108 CFU per dose orally) after Ts infection. (B) Whole heart images of Ts-infected mice (Ts) (n = 6) and Ts-infected mice treated with alive A. muciniphila (Ts+AAm) (n = 6) or pasteurized A. muciniphila (Ts+PAm) (n = 6). Representative images are shown. (C) Heart mass-to-body weight ratio (HM/BW). (D and E) Ejection fraction (EF) and left ventricular weight (LVM) obtained by cardiac ultrasound. (F and G) qPCR analysis of Collagen-1 (Col1) and α-SMA expression in the heart tissue of mice. (H and I) Masson staining and Col1 immunohistochemistry results of heart tissues. Magnification, 200×. Scale bars, 100 μm. Representative images are shown. Data are shown as individual data points and mean ± SD. Data were compared by one-way ANOVA followed by Tukey multiple comparison tests. ns, not significant; *, p < 0.05; **, p < 0.01, ***, p <0.001, ****, p <0.0001.
Fig 6.
TLR2 participated in the protection of PAm against helminth-induced cardiac fibrosis.
(A) Experimental scheme of pasteurized A. muciniphila administration (8 × 108 CFU per dose orally) after Ts infection in wild type (WT) or TLR2 knockout (TLR2 KO) mice. (B) Whole heart images of Ts-infected WT or TLR2 KO mice (Ts) (n = 6) and Ts-infected mice treated with pasteurized A. muciniphila (Ts+PAm) (n = 6). Representative images are shown. (C) Heart mass-to-body weight ratio (HM/BW). (D and E) Ejection fraction (EF) and left ventricular weight (LVM) obtained by cardiac ultrasound. (F and G) qPCR analysis of Collagen-1 (Col1) and α-SMA expression in the heart tissue of mice. (H and I) Masson staining and Col1 immunohistochemistry results of heart tissues. Magnification, 200×. Scale bars, 100 μm. Representative images are shown. Data are shown as individual data points and mean ± SD. Data were compared by one-way ANOVA followed by Tukey multiple comparison tests. ns, not significant; *, p < 0.05; **, p < 0.01, ***, p <0.001, ****, p <0.0001.