NMR-Based Metabolomic Analysis of Sera in Mouse Models of CVB3-Induced Viral Myocarditis and Dilated Cardiomyopathy

Figures

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  Figure 1. Schematic diagram of the experimental design.

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  Figure 2. Representative histopathological images of myocardial tissues derived from the established models of aVMC, cVMC and DCM mice and the normal control mice (CON). (A) Histopathological images of the myocardial tissues; (B) Tissue sections stained with hematoxylin-eosin (×400); (C) Tissue sections stained with Masson (×400).

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  Figure 3. Multivariate analyses for 1D 1H-NMR spectra recorded on sera derived from six groups of mice (CON-w2, CON-w6, CON-w24, aVMC, cVMC, DCM). (A) HCA clustering tree for the six groups; (B) PCA scores plot for the six groups; (C–E) PCA scores plots for aVMC and CON-w2 (C); for cVMC and CON-w6 (D); for DCM and CON-w24 (E). Each point represents a serum sample derived from an individual mouse.

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  Figure 4. OPLS-DA analyses of the mouse sera to identify significant metabolites significantly responsible for distinguishing metabolic profiles between three pathologic groups (aVMC, cVMC, DCM)) and control groups (CON-w2, CON-w6, CON-w24). (A–C) OPLS-DA scores plots; (D–F) the corresponding OPLS-DA loading plots. The red color indicates that the variables are very significant (|r| > 0.482 in (D), |r| > 0.497 in (E), |r| > 0.514 in (F); VIP > 1); orange indicates that the variables are significant (0.349 <|r| < 0.482 in (D), 0.355 <|r| < 0.492 in (E), 0.361 <|r| < 0.514 in (F); VIP > 1); blue indicates that the variables are insignificant (NS).

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  Figure 5. Multivariate ROC analysis of mouse sera for identifying potential biomarkers. (A) Taurine, valine and acetate identified from the ROC analysis of aVMC vs. CON-w2. (B) Glycerol, valine and leucine identified from cVMC vs. CON-w6. (C) Citrate, glycine and isoleucine identified from DMC vs. CON-w24.

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  Figure 6. Significantly disturbed metabolic pathways identified from pathway analyses based on serum levels of metabolites. (A) aVMC vs. CON-w2; (B) cVMC vs. CON-w6; (C) DCM vs. CON-w24. Pathway impact values (PIVs) were calculated from pathway topology analysis, and p values were computed from metabolite set enrichment analysis. Significantly disturbed metabolic pathways were identified with pathway impact values > 0.2 and p values < 0.05, using the Pathway Analysis module provided by MetaboAnalyst 5.0 webserver.