Plumbagin protect against sepsis-induced myocardial injury in mice by inhibiting the JAK2/STAT3 signaling pathway to reduce cardiomyocyte pyroptosis

doi:10.12122/j.issn.1673-4254.2024.11.18

Abstract

Abstract:

Objective To explore the mechanism of plumbagin for protecting against sepsis-induced myocardial injury in mice. Methods Network pharmacology analysis was used to obtain the key targets of plumbagin and diseases, which were subjected to GO and KEGG analysis, and the binding energy was verified using molecular docking. In a mouse model of cecal ligation and puncture (CLP), the protective effect of plumbagin treatment prior to CLP against sepsis-induced myocardial injury was evaluated by examination of myocardial function and pathology using echocardiography and HE staining. Serum levels of CK-MB, LDH, MDA, IL-1β and IL-18 and myocardial ROS level in the mice were detected, and Western blotting was used to determine the protein expression levels of STAT3, GSDMD, caspase-11, JAK2, P-STAT3, P-JAK2, GSDMD-N and HMGB1 in the myocardial tissues. Results Five core targets were screened from the 10 intersecting genes. Molecular docking showed strong binding affinity of plumbagin to STAT3, p-STAT3, and JAK2. Compared with the sham-operated mice, the mouse models of CLP-induced sepsis had significantly decreased CO, LVEF, LVFS and SV and increased serum levels of CK-MB, LDH, MDA and myocardial inflammatory factors and ROS. HE staining and Western blotting showed obvious myocardial injury in the septic mice with increased expressions of JAK2/STAT3 signaling pathway and pyroptosis-related proteins (P<0.05). Pretreatment with plumbagin significantly improved cardiac functions of CLP mice, lowered serum levels of CK-MB, LDH, MDA, inflammatory factors and myocardial ROS, and decreased the expression levels of JAK2/STAT3 signaling pathway and pyroptosis-related proteins. Conclusion Plumbagin pretreatment alleviates myocardial injury in septic mice possibly by inhibiting the STAT3 signaling pathway to reduce cardiomyocyte pyroptosis.

Key words: plumbagin, sepsis induced myocardial injury, pyroptosis, network pharmacology

Ruoli DU, Qi YUN, Yiren WANG, Xinyu DOU, Hongwei YE, Jiahui WANG, Qin GAO. Plumbagin protect against sepsis-induced myocardial injury in mice by inhibiting the JAK2/STAT3 signaling pathway to reduce cardiomyocyte pyroptosis[J]. Journal of Southern Medical University, 2024, 44(11): 2209-2219.

Figures/Tables 11

Fig.1 Wein diagram of plumbagin-sepsis induced myocardial injury-pyroptosis.

Fig.2 Core protein-protein interaction (PPI) network. A: Visual regulatory network of plumbagin-sepsis myocardial injury-pyroptosis. B: PPI network of plumbagin-sepsis myocardial injury-pyroptosis related targets.

Fig.3 GO function analysis results.

Fig.4 Results of KEGG pathway enrichment analysis.

Fig.5 Molecular docking results. A: Molecular docking diagram of plumbagin and STAT3. B: Molecular docking diagram of plumbagin and JAK2. C: Molecular docking diagram of plumbagin and p-STAT3.

Fig.6 Cardiac ultrasound images and cardiac function indexes of the mice in different groups (Mean±SD, n=6). A: Evaluation of cardiac function by M-mode echocardiography. B: Comparison of the echocardiographic parameters (LVEF: Left ventricle ejection fraction; LVFS: Left ventricular fractional shortening). **P<0.01, ***P<0.001 vs Sham group, #P<0.05, ##P<0.01, ###P<0.001 vs CLP group.

Fig.7 HE staining of myocardial tissues of the mice in different groups.

Fig.8 DHE fluorescence staining of myocardial fibers in different groups. A: Fluorescence probe for ROS in the cardiac tissues (Original magnification: ×200). B: ROS fluorescence intensity in each group (Mean±SD, n=6). ***P<0.001 vs sham group; ##P<0.01, ###P<0.001 vs the CLP group.

Fig.9 Serum IL-18 (A) and IL-1β (B) levels in the myocardial tissue in different groups (Mean±SD, n=6). ***P<0.001 vs sham group; #P<0.05 vs CLP group.

Fig.10 Serum MDA (A), CK-MB (B) and LDH (C) levels in the myocardial tissue in different groups (Mean±SD, n=6). ***P<0.001 vs sham group; #P<0.05, ##P<0.01 ###P<0.001 vs CLP group.

Fig.11 Comparison of GSDMD、Caspase-11、HMGB1、P-JAK2、JAK2、P-STAT3、STAT3 and GSDMD-N protein levels in the myocardial tissues among the 4 groups. A-C: Protein bands in Western blotting. D-I: Relative protein levels of GSDMD、Caspase-11、HMGB1、P-JAK2、JAK2、P-STAT3、STAT3 and GSDMD-N proteins (Mean±SD, n=3). *P<0.05, **P<0.01, ***P<0.001 vs sham group; #P<0.05, ##P<0.01, ###P<0.001 vs CLP group.

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