Xionggui Decoction alleviates heart failure in mice with myocardial infarction by inhibiting oxidative stress-induced cardiomyocyte apoptosis

doi:10.12122/j.issn.1673-4254.2024.07.22

Abstract

Abstract:

Objective To explore the protective effect of Xionggui Decoction against cardiac myopathy in a mouse model of heart failure following myocardial infarction (MI) and explore the underlying mechanism. Methods We searched TCMSP, GeneCards, and CTD databases for the targets of active ingredients Xionggui Decoction and heart failure, and the intersecting targets were analyzed with GO and KEGG pathway enrichment analysis using DAVID database. In a mouse model of heart failure following acute MI induced by coronary artery ligation, the cardiac protective effects of 3 g/kg Xionggui Decoction were evaluated by assessing cardiac function, cardiac myopathy and ventricular remodeling of the mice using HE staining, Masson staining, RT-qPCR, and immunohistochemistry. We also tested the effect of Xionggui Decoction at 50 and 100 μg/mL on tert-butylhydrogen peroxide (TBHP)-induced apoptosis of H9C2 cells using CCK8 assay, detection kits for ROS, MDA, SOD, JC-1 and Hoechst 33342/PI staining. Results Network pharmacological analysis identified 62 potential targets of Xionggui Decoction for treatment of heart failure, and the core targets included PTGS2, ESR1, caspase-3, PPARG, HSP90AA1, BCL2, JUN, and GSK3B, which were involved in cell apoptosis and the AGE-RAGE, P53, PI3K-Akt, and VEGF signaling pathways. In the mouse models of heart failure, treatment with Xionggui Decoction significantly alleviated cardiac myopathy and ventricular remodeling, obviously improved heart function of the mice, lowered myocardial expressions of caspase-3 and BAX, and enhanced the expression of BCL2. In H9C2 cells, Xionggui Decoction significantly alleviated TBHP-induced cell apoptosis by inhibiting oxidative stress in the cells. Conclusion Xionggui Decoction can alleviate myocardial injury and improve cardiac function in mice with heart failure following acute MI possibly by inhibiting cardiomyocyte apoptosis induced by oxidative stress.

Key words: Xionggui Decoction, heart failure, network pharmacology, apoptosis, oxidative stress

Zhijun REN, Jianxin DIAO, Yiting WANG. Xionggui Decoction alleviates heart failure in mice with myocardial infarction by inhibiting oxidative stress-induced cardiomyocyte apoptosis[J]. Journal of Southern Medical University, 2024, 44(7): 1416-1424.

Figures/Tables 9

Fig.2 GO and KEGG pathway enrichment analysis of the potential targets.

Fig.3 Xionggui Decoction improves heart function of the mice with heart failure. A: Left ventricular end-systolic dimension (LVEDs). B: Left ventricular end-diastolic dimension (LVEDd). C: Left ventricular ejection fraction (LVEF). D: Left ventricular fractional shortening (LVFS). ##P<0.01 vs Sham group; *P<0.05, **P<0.01 vs Model group.

Fig.4 Xionggui Decoction alleviates myocardial damage in mice with heart failure. A: Heart weight/body weight ratio. B-D: Serum LDH, CK, and AST levels. ##P<0.01 vs Sham group; *P<0.05, **P<0.01 vs Model group.

Fig.5 Xionggui Decoction alleviates myocardial necrosis and inflammation in mice with heart failure. The green arrows indicate the inflammatory cells (scale bar: 100 μm).

Fig.6 Xionggui Decoction alleviates myocardial fibrosis in mice with heart failure. A: Masson staining of the myocardial tissue. B: Collagen volume fraction. C: Relative expression of Collagen I. D: Relative expression of Collagen III. ##P<0.01 vs Sham group; **P<0.01 vs Model group.

Fig.7 Xionggui Decoction alleviates myocardial cell apoptosis in mice with heart failure. A: Immunohistochemical analysis. B: Relative expression of BAX. C: Relative expression of BCL2. D: Relative expression of caspase-3. #P<0.05, ##P<0.01 vs Sham group; **P<0.01 vs Model group.

Fig.8 Xionggui Decoction alleviate H9C2 cell injury induced by oxidative stress. A: Viability of H9C2 cells incubated with different concentrations of Xionggui Decoction (XGT). B: Viability of H9C2 cells incubated with different concentrations of TBHP. C: Viability of H9C2 cells incubated with different concentrations of XGT (50 and 100 µg/mL), followed by treatment with TBHP (200 μmol/L). ##P<0.01 vs Control group; *P<0.05, **P<0.01 vs TBHP group.

Fig.9 Xionggui Decoction protects H9C2 cells from TBHP-induced oxidative stress. A: Reactive oxygen species (ROS) levels. B: Malondialdehyde (MDA) content. C: Superoxide dismutase (SOD) activity. ##P<0.01 vs Control; *P<0.05 vs TBHP; **P<0.01 vs TBHP.

Fig.10 Xionggui Decoction alleviates TBHP-induced H9C2 cell apoptosis. A: Hoechst 33342/PI fluorescent staining. B: JC-1 staining. ##P<0.01 vs Control; **P<0.01 vs TBHP.

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