Levocarnitine improves cardiac energy metabolic remodeling in myocarditis mice

Figures

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  FIGURE 1. Workflow of the study based on network pharmacology and molecular docking aimed at exploring the mechanism of levocarnitine in treating myocarditis.

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  FIGURE 2. LC improves cardiac insufficiency and reduces myocardial inflammatory infiltration in EAM mice. (A) Changes in body weight and (B) body weight of mice in each group on day 21, n = 3. (C) Parasternal left ventricular short-axis M-mode echocardiograms of mice in each group on day 21. (D) Changes in left ventricular ejection fraction and (E) left ventricular shortening fraction, n = 3. (F) Myocardial inflammatory infiltration scores in each group of mice, n = 3. (G) HE staining showing inflammatory lesions in cardiac tissue (×200). Data are expressed as Mean ± SEM with individual data points shown. LVEF, left ventricular ejection fraction; FS, fractional shortening.

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  FIGURE 3. LC attenuated myocardial mitochondrial structural damage and respiratory chain abnormalities in EAM mice. (A) Transmission electron microscopy showing morphological changes in mouse heart mitochondria (×5,000, scale bar = 2 μm). Changes in (B) mitochondrial complex IV activity and (C) ATP levels in each group, n = 3. (D,E) DCFH-DA staining flow cytometry to detect ROS levels in each group, n = 3. Data are expressed as Mean ± SEM with individual data points shown. ATP, adenosine triphosphate; ROS, reactive oxygen species.

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  FIGURE 4. LC improves myocardial mitochondrial substrate utilization in EAM mice. (A) Serum H-FABP levels in each group of mice, n = 3. (B–D) Western blot analysis of OCTN-2 and CPT-1B proteins, n = 3. (E) FFA and (F) LAC levels in myocardial tissue from each group of mice, n = 3. Data are expressed as Mean ± SEM with individual data points shown. FABP, heart-type fatty acid-binding protein; OCTN2, organic carnitine transporter novel type 2; CPT-1B, carnitine palmitoyltransferase-1B; FFA, free fatty acid; LAC, lactic acid.

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  FIGURE 5. Network pharmacology and enrichment analysis of levocarnitine for myocarditis. (A) Venn diagram showing overlapping targets linked to levocarnitine and myocarditis, and (B) PPI diagram of potential targets of levocarnitine for myocarditis. (C) GO functional enrichment analysis. (D) KEGG pathway enrichment analysis. The closer the color is to red, the smaller the P value, indicating more significant enrichment.

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  FIGURE 6. Docking model of LC with AKT1. The yellow region represents the ligand LC, while the purple region corresponds to the AKT1 protein.

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  FIGURE 7. Effect of LC on Akt and PGC-1α in EAM mice. (A–C) Changes in Akt, p-Akt, and PGC-1α were analyzed by Western blots, n = 3. Data are expressed as Mean ± SEM with individual data points shown. PGC-1α, peroxisome proliferator-activated receptor gamma coactivator-1α.

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  FIGURE 8. Pearson’s correlation analysis of p-Akt and PGC-1α with mitochondrial metabolic function. Scatter plots showing the correlation between myocardial p-Akt protein expression and (A) COX IV activity, (B) CPT-1B expression, (C) ROS levels, and (D) ATP. Scatter plots showing the correlation between myocardial PGC-1α protein expression and (E) COX IV activity, (F) CPT-1B expression, (G) ROS levels, and (H) ATP. (n = 9 from 3 groups).