Esmolol Alleviates Sepsis‐Associated Myocardial Injury by Acting on the APAF1 and JAK2/STAT3 Inflammatory Pathways

Figures

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  Figure 1. Screening and validation of core targets of esmolol in SAMI. (A) Venn diagram showing the overlap between SAMI‐related DEGs and esmolol target proteins. (B) GO/KEGG enrichment analysis of the 51 overlapping genes. (C) Volcano plot of SAMI‐related DEGs, highlighting core candidate genes. (D) The external dataset GSE243217 shows the expression of four targets with consistent trends. Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01.

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  Figure 2. Molecular docking of esmolol with key targets and CETSA validation. (A) Molecular docking of esmolol with APAF1 and JAK2 (binding affinities: −7.73 and −6.05 kcal/mol, respectively). The interaction bonds are shown on the right; (B) CETSA validation of the direct binding of esmolol to APAF1 in cells. Data are presented as mean ± SEM. ns, not significant; ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

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  Figure 3. Esmolol alleviates LPS‐induced injury, apoptosis, and oxidative stress in H9C2 cardiomyocytes. (A) CCK‐8 assay for cell viability (n = 6); (B–C) Flow cytometry analysis of apoptosis rates; (D) representative image of TUNEL staining; (E) flow cytometry analysis of intracellular ROS levels; (F) kit‐based assay for MDA levels; (G) flow cytometry profile of JC‐1 staining (n = 3). Data are presented as mean ± SEM. ∗∗∗∗ p < 0.0001.

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  Figure 4. Esmolol inhibits LPS‐induced apoptosis in H9C2 cells and the activation of the JAK2/STAT3 inflammatory pathway (A) Western blot analysis of APAF1 and the apoptosis pathway; (B) ELISA analysis of proinflammatory cytokines; (C) Western blot analysis of the JAK2/STAT3 pathway (n = 3). Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

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  Figure 6. Esmolol improves cardiac function, myocardial pathological damage, oxidative stress, and inflammatory responses in rats with sepsis. (A) Representative images of rat echocardiography and quantitative analysis of cardiac function parameters (HR, EF, FS, LVEDD, LVESD) (n = 3); (B) representative images of HE‐stained rat myocardial tissue (n = 3); (C) detection of ATP content, MDA content, and SOD activity in rat myocardial tissue (n = 6); (D) detection of myocardial injury markers (LDH, CK‐MB, cTnI, cTnT) in rats (n = 6); (E) detection of myocardial inflammatory factors (IL‐6, TNF‐α, IL‐1β) in rats (n = 6). Data are presented as mean ± SEM. ns, not significant; ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001.

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  Figure 7. Esmolol exerts cardioprotective effects by regulating the APAF1 apoptotic pathway and the JAK2/STAT3 inflammatory pathway. (A) Expression of APAF1 and apoptosis‐related proteins in rat myocardial tissue (n = 3); (B) detection of JAK2/STAT3 phosphorylation levels in rat myocardial tissue (n = 3). Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.