The Impact of Sleep Disturbance on Gut Microbiota, Atrial Substrate, and Atrial Fibrillation Inducibility in Mice: A Multi-Omics Analysis

Figures

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  Figure 1. Enhanced AF inducibility, atrial lipid accumulation, and fibrosis in SD mice. (A) Representative image of surface ECG recordings during burst pacing in control and SD group. AF inducibility (B), AF susceptibility (C), and AF duration (D) in the control (n = 6) and SD group (n = 6). Chi-square test. (E) Representative Masson trichrome, Sirius Red, and Oil red O staining for fibrosis disarray, collagen deposition, and lipid droplets in the atrium. (F) Representative Western blot and the expression levels of collagen I and α-SMA in mouse atria of the two groups. Data are presented as the mean ± SEM. **, p < 0.01;***, p < 0.001; n = 3 for each group, Student’s t-test.

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  Figure 2. Altered gut microbiota in SD mice. (A) The ASVs accumulation curve. The observed OTUs (B), Shannon index (C), and Chao1 richness (D) in the control and SD group. Boxes represent the interquartile ranges; the inside line or points represent the median, and circles are outliers, Kruskal–Wallis test. (E) Principal coordinates analysis (PCoA) based on Bray–Curtis distance. The green circles represent control, and the purple circles denote SD. (F) Phylum-level taxonomic abundance and proportion for the control (left) and SD (right) groups; different taxa are differentiated by color. n = 8 for each group. (G) The ratio of the Firmicutes to Bacteroidetes (F/B ratio) in the control and SD groups. (H) Differential enriched genera between control and SD group, t-test. (I) The cladogram of differential enriched genera. (J) Differential enriched functional pathways between control and SD group, t-test.

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  Figure 3. Changed metabolic patterns in plasma of SD mice. (A) Score scatter plots of orthogonal PLS-DA (OPLS-SA) comparing the plasma metabolic differences show the separation between control and SD in ES+ and ES−. ES+, positive ion mode; ES−, negative ion mode. (B) Volcano plots for the alterations of plasma metabolites between ES+ and ES− groups. Metabolite at p < 0.05 and variable importance (VIP) in the projection threshold of >1 was significantly different. Downregulation and upregulation were determined by fold-change. (C) Pie chart showing the proportion of metabolite annotated to the classification of superclass based on the metabolomics workbench. (D) Bubble analysis of metabolic KEGG pathways between control and SD group. (E) Circos plots showed Spearman’s correlation analysis (p < 0.05) between phyla and metabolites classified based on the superclass. Red indicates a positive correlation, and blue indicates a negative correlation. Each phylum and metabolic superclass were seen in a different color.

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  Figure 4. Modified transcriptional signatures in atria of SD mice. (A) Principal coordinates analysis (PCoA) based on Bray–Curtis distance. The green circles represent control and the purple circles denote SD. (B) Venn plot showed the mRNA detected in control and SD. (C) Volcano plots showing the results of DESeq2-based differential genes analyses. Adjusted p-value < 0.05 and |log2 (FoldChange)| > 1 were set as the threshold for significantly differential expression. Information of NES (D) and GeneRatio (E) of KEGG pathways revealed by the Gene Set Enrichment Analysis (GSEA) and the network (F) and ridge plot (G) of five metabolism-related pathways.