Integrated Metabolomics and Proteomics Analysis of the Myocardium in a Mouse Model of Acute Viral Myocarditis

Figures

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  Figure 1. Overall experimental procedures and assessment of myocardial inflammation. (A) Schematic illustration showing the design of animal experiments. (B) Gross examination of the hearts from Control and AVMC mice on Day 7 postinfection. (C) Comparison of cardiac pathological scores between the two groups. Each dot represents an individual mouse. (D) Representative HE staining images of heart sections from the two groups (scale bar = 500 μm, 100 μm). **p value < 0.01.

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  Figure 2. Altered myocardial metabolomic profiles in AVMC mice. (A) 3D scatter plot of PCA results. (B) OPLS‐DA analysis showing good discrimination between the Control and AVMC groups (R2X = 0.575, R2Y = 0.979, Q2Y = 0.954). (C) The permutation test (n = 200) for the OPLS‐DA model. (D) Volcano plot of DMs between the two groups. DMs were screened based on the criterion of VIP > 1.0 and p value < 0.05. Blue dots indicate downregulated metabolites, and red dots indicate upregulated metabolites. (E) Hierarchical clustering heat map analysis of DMs at the MS2 level between the two groups.

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  Figure 3. Pathways associated with myocardial DMs. (A) KEGG pathway enrichment analysis of DMs. The significantly enriched pathways are displayed by a bar plot (p value < 0.05), with the bar length indicating the number of enriched metabolites and the bar color indicating the ‐log10(p value). (B) Summary of pathway analysis using MetPA (top 25). The color and size of each circle are based on ‐log10(p value) and pathway impact value, respectively. Statistically significant pathways are annotated with their names in blue font (p value < 0.05 and impact value > 0.2).

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  Figure 4. Altered myocardial proteomics profiles in AVMC mice. (A) Venn diagram of proteins identified in the Control and AVMC groups. (B) 3D PCA analysis of all samples based on their protein expression. (C) Volcano plot of DPs between the two groups. DPs were screened based on the criterion of |Fold Change| > 1.5 and p value < 0.05. Blue dots indicate downregulated proteins, and red dots indicate upregulated proteins. (D) Heatmap of DPs using unsupervised hierarchical clustering.

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  Figure 5. GO and KEGG enrichment analyses of DPs. (A) GO enrichment terms for the upregulated and downregulated DPs were grouped into three main ontologies: BP, CC, and MF. (B) KEGG pathway functional classification and annotation of all DPs. The number shown on the right side of each bar represents the number of proteins.

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  Figure 6. Integrated metabolomics and proteomics pathway analysis. (A) Venn diagram of the pathways in which DMs and DPs were involved. (B) The most enriched shared KEGG pathways were shown on the graph based on the p value of DMs < 0.05.