Transforming growth factor-β3 attenuates septic cardiomyopathy by reversing cardiomyocyte metabolic reprogramming through Smad7 signaling

Figures

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  Fig. 1. Myocardial tissue injury accompanied by metabolic changes in SCM. A Representative H&E staining and pathological damage scores of myocardial tissues from SCM rats at different time points. Each group n = 9. Scale bar was 50 μm. Differences were assessed using one-way ANOVA and Tukey’s multiple comparison test. B ELSIA was performed to detect lactate levels in myocardial tissues of SCM rats at different time points. Each group n = 3. Differences were assessed using one-way ANOVA and Tukey’s multiple comparison test.

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  Fig. 2. TGF-β3 showed protective effects against SCM. A Representative H&E staining and pathological damage scores of rat myocardial tissue. Each group n = 3. Scale bar was 20 μm. Differences were assessed using one-way ANOVA and Tukey’s multiple comparison test. B TUNEL staining was used to detect cardiomyocyte apoptosis. Each group n = 3. Differences were assessed using one-way ANOVA and Tukey’s multiple comparison test. C Transmission electron microscopy was used to detect mitochondrial damage. Each group n = 3. Scale bars are 2 μm above and 500 nm below. red arrows point to mitochondria. Differences were assessed using one-way ANOVA and Tukey’s multiple comparison test.

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  Fig. 3. TGF-β3 reversed cardiomyocyte metabolic reprogramming in SCM. A The extracellular acidification rate (ECAR) of cardiomyocytes was detected by seahorse. Each group n = 3. Differences were assessed using one-way ANOVA and Tukey’s multiple comparison test. B Oxygen consumption rate (OCR) of cardiomyocytes was assayed by seahorse. Each group n = 3. Differences were assessed using one-way ANOVA and Tukey’s multiple comparison test.

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  Fig. 4. TGF-β3 promotes Smad7 phosphorylation and inhibits SKP1-mediated ubiquitination degradation of Smad7. A Western blot was performed to detect the total protein and phosphorylation levels of TGF-β3, SKP1, Smad7 and Smad2/3 in myocardial tissues of SCM rats. Each group n = 3. and subsequently analyzed quantitatively by image lab software. Differences were assessed using one-way ANOVA and Tukey’s multiple comparison test. B Western blot was performed to detect the total protein and phosphorylation levels of TGF-β3, Smad7 and Smad2/3 in SCM primary rat cardiomyocytes. Each group n = 3. and subsequently analyzed quantitatively by image lab software. Differences were assessed using one-way ANOVA and Tukey’s multiple comparison test. C Immunoprecipitation assay to detect ubiquitinated degradation of Smad7 in cardiomyocytes.