Diet Alters Serum Metabolomic Profiling in the Mouse Model of Chronic Chagas Cardiomyopathy

Figures

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  Figure 1. T. cruzi infection altered body weights and glucose levels in chronic mice fed on different diets. (a) T. cruzi-infected HFD mice displayed greater body weight compared to infected RD-fed mice at 150 DPI. (b) T. cruzi infection increased the weights of the hearts compared to uninfected mice irrespective of the diets fed at 150 DPI. (c) The weights of the livers of HFD-fed mice (both uninfected and infected) were significantly increased compared to RD-fed (both uninfected and infected) mice. T. cruzi infection showed no significant effect on the weights of the livers. (d) Serum glucose measurements demonstrated that chronic T. cruzi infection has no significant effect on the levels of fasting glucose in RD-fed mice. However, the levels of fasting glucose significantly increased in infected HFD mice compared to infected RD-fed mice. (e) Infected HFD mice showed a better clearance of glucose during OGTT analysis compared to infected RD mice even though the basal levels of glucose were significantly greater in infected HFD mice compared to infected RD mice. The error bars represent standard error of the mean. ∗p ≤ 0.05, ∗∗p ≤ 0.01, or ∗∗∗p ≤ 0.001 compared to uninfected RD mice. #p ≤…

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  Figure 2. Immunoblot analysis of the livers demonstrated disrupted lipid metabolism in T. cruzi-infected mice and HFD further enhanced interruption of hepatic lipid metabolism during chronic Chagas disease. (a) T. cruzi-infected HFD-fed mice showed significantly reduced hepatic lipid metabolism compared to infected RD-fed mice as demonstrated by immunoblot analysis probed for various lipid metabolism markers such as fatty acid synthase (FAS), phosphorylated ATP-citrate lyase (pACL), phosphorylated acetyl co A carboxylase (p-ACC), carnitine palmitoyltransferase I (CPT1), and Lipin 1. Infected mice showed significantly increased hepatic levels of PPARα, a regulator of fatty acid oxidation compared to uninfected mice, and HFD further enhanced the levels of PPARα in the livers of infected mice. (b–h) Fold changes in the protein levels of FAS, pACL, acetyl CoA-synthase (AceCS1), pACC, CPT1, Lipin 1, and PPARα were normalized to GDI expression and represented as the bar graphs (b–h, respectively). The error bars represent the standard error of the mean. ∗p ≤ 0.05, ∗∗p ≤ 0.01, or ∗∗∗p ≤ 0.001 compared to uninfected RD mice. #p ≤ 0.05, ##p ≤ 0.01, or ###p ≤ 0.001 compared to infected RD mice. ^p ≤…

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  Figure 3. Comparison of amino acid profiling between T. cruzi-infected and uninfected mice fed on different diets at 150 DPI. (a) Heat map representing the serum levels of various amino acids during chronic infection in mice fed on either a RD or a HFD compared to uninfected RD mice at 150 DPI. Each row represents data for a specific metabolite, and each column represents the RD-fed uninfected (RD), RD-fed infected (RDI), HFD-fed uninfected (HFD), or HFD-fed infected (HFDI) mouse group. Different colors correspond to the different intensity level of metabolites (red > the control RD group and green < the control RD group). Statistically significant differences were tested using Student's t-test, and the corrected p values (q-value) are presented next to the heat map. Red, yellow, and green rectangles indicate high, moderate, and low expressions, respectively. (b) The percentage changes in the levels of amino acid metabolites in infected mice compared to the respective diet-fed (RD and HFD) uninfected mice represented by a bar graph (∗significant change).

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  Figure 4. Comparison of biogenic amines profiling between T. cruzi-infected and uninfected mice fed on different diets at 150 DPI. (a) Heat map representing the serum levels of various biogenic amines during chronic infection in mice fed on either a RD or a HFD compared to uninfected RD mice at 150 DPI. Each row represents data for a specific metabolite, and each column represents the RD-fed uninfected (RD), RD-fed infected (RDI), HFD-fed uninfected (HFD), or HFD-fed infected (HFDI) mouse group. Different colors correspond to the different intensity level of metabolites (red > the control RD group and green < the control RD group). Statistically significant differences were tested using Student's t-test, and the corrected p values (q-value) are presented next to the heat map. Red, yellow, and green rectangles indicate high, moderate, and low expressions, respectively. (b) The percentage changes in the levels of biogenic amines in infected mice compared to the respective diet-fed (RD and HFD) uninfected mice represented by a bar graph (∗significant change).

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  Figure 5. Comparison of acylcarnitine profiling between T. cruzi-infected and uninfected mice fed on different diets at 150 DPI. (a) Heat map representing the serum levels of various acylcarnitines during chronic infection in mice fed on either a RD or a HFD compared to uninfected RD mice at 150 DPI. Each row represents data for a specific metabolite, and each column represents the RD-fed uninfected (RD), RD-fed infected (RDI), HFD-fed uninfected (HFD), or HFD-fed infected (HFDI) mouse group. Different colors correspond to the different intensity level of metabolites (red > the control RD group and green < the control RD group). Statistically significant differences were tested using Student's t-test, and the corrected p values (q-value) are presented next to the heat map. Red, yellow, and green rectangles indicate high, moderate, and low expressions, respectively. (b) The percentage changes in the levels of acylcarnitines in infected mice compared to the respective diet-fed (RD and HFD) uninfected mice represented by a bar graph (∗significant change).

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  Figure 6. Comparison of sphingolipid profiling between T. cruzi-infected and uninfected mice fed on different diets at 150 DPI. (a) Heat map representing the serum levels of various sphingolipids during chronic infection in mice fed on either a RD or a HFD compared to uninfected RD mice at 135 DPI. Each row represents data for a specific metabolite, and each column represents the RD-fed uninfected (RD), RD-fed infected (RDI), HFD-fed uninfected (HFD), or HFD-fed infected (HFDI) mouse group. Different colors correspond to the different intensity level of metabolites (red > the control RD group and green < the control RD group). Statistically significant differences were tested using Student's t-test, and the corrected p values (q-value) are presented next to the heat map. Red, yellow, and green rectangles indicate high, moderate, and low expressions, respectively. (b) The percentage changes in the levels of sphingolipid metabolites in infected mice compared to the respective diet-fed (RD and HFD) uninfected mice represented by a bar graph (∗significant change).

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  Figure 7. Comparison of 14 lysophosphatidylcholines profiling between T. cruzi-infected and uninfected mice fed on different diets at 150 DPI. (a) Heat map representing the serum levels of various lysophosphatidylcholines during chronic infection in mice fed on either a RD or a HFD compared to uninfected RD mice at 150 DPI. Each row represents data for a specific metabolite, and each column represents the RD-fed uninfected (RD), RD-fed infected (RDI), HFD-fed uninfected (HFD), or HFD-fed infected (HFDI) mouse group. Different colors correspond to the different intensity level of metabolites (red > the control RD group and green < the control RD group). Statistically significant differences were tested using Student's t-test, and the corrected p values (q-value) are presented next to the heat map. Red, yellow, and green rectangles indicate high, moderate, and low expressions, respectively. (b) The percentage changes in the levels of LPC metabolites in infected mice compared to the respective diet-fed (RD and HFD) uninfected mice represented by a bar graph (∗significant change).

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  Figure 8. Comparison of phosphocholine (73) profiling between T. cruzi-infected and uninfected mice fed on different diets at 150 DPI. (a, b) Heat map representing the serum levels of various glycerol-phospholipids that included (a) diacyl PC and (b) acyl-alkyl PC during chronic infection in mice fed on either a RD or a HFD compared to uninfected RD mice at 150 DPI. Each row represents data for a specific metabolite, and each column represents the RD-fed uninfected (RD), RD-fed infected (RDI), HFD-fed uninfected (HFD), or HFD-fed infected (HFDI) mouse group. Different colors correspond to the different intensity level of metabolites (red > the control RD group and green < the control RD group). Statistically significant differences were tested using Student's t-test, and the corrected p values (q-value) are presented next to the heat map. Red, yellow, and green rectangles indicate high, moderate, and low expressions, respectively. (c, d) The percentage changes in the levels of phosphocholine metabolites that included (c) diacyl PC and (d) acyl-alkyl PC in infected mice compared to the respective diet-fed (RD and HFD) uninfected mice represented by a bar graph (∗significant change).