Gemfibrozil mitigates caspase-11-driven myocardial pyroptosis in ischemia/reperfusion injury in mice

Figures

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  Fig. 1. Cleaved caspase-11 and GSDMD-N levels in the ischemic region at 0, 3, 6, 12, 24, and 48 h after reperfusion. (A) Scheme of the experimental design. (B) CK-MB activity. (C) Representative western blot images of caspase-11 and GSDMD. (D) Quantification of cleaved caspase-11 levels. (E) Quantification of GSDMD-N levels. Data are presented as the mean ± standard deviation of six animals. The experimental data were analyzed using Dunnett's method. *p < 0.05 vs. the control group.

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  Fig. 2. Effects of gemfibrozil (Gem) on caspase-11 and GSDMD levels in the ischemic region of mice 48 h after ischemia/reperfusion (I/R) (black plots) and Sham (white plots). (A) Stick model and (B) surface model from molecular docking simulations showing Gem binding to caspase-11. (C) Scheme of the experimental design. (D) Representative western blot images of caspase-11 and GSDMD. (E) Quantification of cleaved caspase-11 levels. (F) Quantification of GSDMD-N levels. Data are presented as the mean ± standard deviation of six animals. The experimental data were analyzed using Tukey's method. *p < 0.05 vs. the Sham group. #p < 0.05 vs. the vehicle-treated (0 μmol/L) group 48 h after I/R (IR48h).

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  Fig. 3. Effects of gemfibrozil (Gem) on infarct size and cardiac function 48 h after ischemia/reperfusion (I/R) (black plots) and Sham (white plots). (A) Representative Evans Blue (EB)/TTC-stained images showing infarct areas. (B) AAR/LV ratio (area at risk as a percentage of the left ventricle). (C) IA/AAR ratio (infarct area as a percentage of the area at risk). (D) Representative M-mode echocardiographic images. (E) Left ventricular internal diameter in diastole (LVIDd). (F) Left ventricular internal diameter in systole (LVIDs). (G) Fractional shortening. (H) Ejection fraction. (I) Representative immunohistochemical images showing GSDMD (red), FITC-WGA (green), and nuclei (blue) in ischemic regions. Data are presented as the mean ± standard deviations of six animals. The experimental data were analyzed using Tukey's method. *p < 0.05 vs. the Sham group. #p < 0.05 vs. the vehicle-treated (0 μmol/L) group 48 h after I/R (IR48h). (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

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  Fig. 4. Effects of gemfibrozil (Gem) on hypoxia/reoxygenation (H/R)-induced necrotic cell death in cultured adult mouse cardiomyocytes. (A) Scheme of the experimental design. (B) Representative images of cardiomyocytes stained with ethidium homodimer III (EthD-III, red), annexin V (green), and Hoechst 33342 (blue). (C) Percentage of EthD-III-labeled necrotic cells among total cells. (D) Lactate dehydrogenase (LDH) activity in cultured supernatants. Data are presented as the mean ± standard deviations of four independent experiments. The experimental data were analyzed using Tukey's method. *p < 0.05 vs. the Normoxia (Nor, white plot) group. #p < 0.05 vs. the vehicle-treated (0 μmol/L) H/R group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

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  Fig. 5. Effects of gemfibrozil (Gem) on caspase-11 and GSDMD levels in cultured adult mouse cardiomyocytes after hypoxia/reoxygenation (H/R). (A) Representative western blot images of caspase-11 and GSDMD. (B) Quantification of cleaved caspase-11 levels. (C) Quantification of GSDMD-N levels. (D) Representative immunohistochemical images showing GSDMD (red) and FITC-WGA (green) in cultured cardiomyocytes. Data are presented as the mean ± standard deviation of four independent experiments. The experimental data were analyzed using Tukey's method. *p < 0.05 vs. the Normoxia (Nor, white plot) group. #p < 0.05 vs. the vehicle-treated (0 μmol/L) H/R group. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)