Allicin ameliorates acute myocardial infarction in rats by modulating calcium homeostasis in cardiomyocytes through the induction of hydrogen sulfide production

Figures

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  FIGURE 1. Allicin improved cardiac function in acute myocardial infarction model rats. (A‒G) Statistical scatter plots of the left ventricular (LV) internal diameter and thickness of the anterior wall at end-systole (LVID s, LVAW s) and end-diastole (LVID d, LVAW d), ejection fraction (EF), fractional shortening (FS), and stroke volume (SV) in the different groups. (H) Representative images of ultrasonic function in each group. Statistical analysis was performed by one-way ANOVA and Tukey’s honestly significant difference (HSD) post hoc test, and the data are expressed as the mean ± SD (n = 8–11). *p < 0.05, **p < 0.01 vs. Sham; △ p < 0.05, △△ p < 0.01 vs. Model; # p < 0.05, ## p < 0.01 vs. 14 mg/kg allicin.

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  FIGURE 2. Allicin reduced the myocardial necrosis area and serum myocardial enzyme levels in acute myocardial infarction rats. (A,B) Percentage of myocardial necrosis area relative to the ventricular and total heart areas, respectively. (C,D) Myocardial enzyme (cTnI, LDH) levels in the serum of acute myocardial infarction rats. (E) Representative myocardial necrosis areas in each group; scale bar = 10 mm. The area circled in green represents the area of myocardial necrosis. Statistical analysis was performed by one-way ANOVA, Tukey’s honestly significant difference (HSD) post hoc test and Tamhane post hoc test, and the data are expressed as the means ±SDs (n = 7–11). *p < 0.05, **p < 0.01 vs. Sham; △ p < 0.05, △△ p < 0.01 vs. Model; # p < 0.05, ## p < 0.01 vs. 14 mg/kg allicin.

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  FIGURE 3. Allicin increased H2S levels and H2S synthase (CBS, CSE) in myocardial tissue and serum. (A,B) H2S levels in the serum and myocardial tissues of rats in each group. (C,D) H2S synthase (CBS, CSE) levels in the serum of rats in each group. (E,F) Fluorescence images of representative CBS and CSE in myocardial tissue from each group. Scale bar = 40 µm. Statistical analysis was performed by one-way ANOVA, Tukey’s honestly significant difference (HSD) post hoc test and Tamhane post hoc test, and the data are expressed as the means ±SDs (n = 10). *p < 0.05, **p < 0.01 vs. Sham; △ p < 0.05, △△ p < 0.01 vs. Model; ## p < 0.01 vs. 14 mg/kg allicin.

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  FIGURE 4. Allicin improved the pathological morphology of the myocardium in acute myocardial infarction rats. H&E staining, magnification ×200. Scale bar = 100 µm (n = 4).

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  FIGURE 5. Allicin regulated cardiomyocyte contraction kinetics and cardiomyocyte calcium transient amplitude in acute myocardial infarction rats. (A) Shortening amplitude; (B) Peak time; (C) Systolic T50; (D) Diastolic T50; (E) Calcium transient amplitude; (F) calcium decline time constant; (G) Maximum ascending velocity; (H) Maximum descending velocity. Statistical analysis was performed by the Kruskal‒Wallis test, and the data are presented as the mean ± SD (n = 29–46 cardiomyocytes from four rats). *p < 0.05, **p < 0.01 vs. Sham; △ p < 0.05, △△ p < 0.01 vs. Model; # p < 0.05, ## p < 0.01 vs. 14 mg/kg allicin. Abbreviations: T50, half-life of decay.

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  FIGURE 6. Allicin regulated calcium transport in cardiomyocytes from acute myocardial infarction rats. (A) NCX calcium removal function, TauNCX (n = 15–34 cardiomyocytes from four rats); (B) SERCA calcium reuptake function, TauSERCA (n = 15–34 cardiomyocytes from four rats); (C) Sarcoplasmic reticulum calcium content, △F/F0 (n = 15–34 cardiomyocytes from four rats); (D) Calcium leakage levels, △Fleak/F0 (n = 22–123 cardiomyocytes from four rats). Statistical analysis was performed by the Kruskal‒Wallis test, and the data are presented as the means ±SDs. * p < 0.05, ** p < 0.01 vs. Sham; △ p < 0.05, △△ p < 0.01 vs. Model; # p < 0.05, ## p < 0.01 vs. 14 mg/kg allicin.

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  FIGURE 7. Allicin regulated the expression of key calcium transport proteins in the myocardial tissues of acute myocardial infarction rats. (A) Representative immunoblots of myocardial tissues for p-RyR2, total RyR, NCX-1, SERCA2a, p-CaMKII, CaMKII and GAPDH obtained from rats. (B–I) Quantitative analyses of total CaMKII, p-CaMKII, SERCA2a, NCX-1, total RyR, p-RyR2 normalized to GAPDH, the ratio of p-CaMKII/total CaMKII (D) and p-RyR2/total RyR (I). Statistical analysis was performed by one-way ANOVA, Tukey’s honestly significant difference (HSD) post hoc test and Tamhane post hoc test, and the data are presented as the means ±SDs (n = 4). *p < 0.05, **p < 0.01 vs. Sham; △ p < 0.05, △△ p < 0.01 vs. Model; # p < 0.05, ## p < 0.01 vs. 14 mg/kg allicin.