Cardioprotective effects of the jiming formula on myocardial metabolism in Mice with myocardial infarction via the AMPK/SIRT1/PGC-1α pathway.

BACKGROUND

Cardiac dysfunction continues to represent a major global health burden, significantly impacting both disease prevalence and survival rates across populations. Mitochondrial dysfunction is a severe pathological characteristic of heart failure. Altered energy metabolism is intimately linked to the advancement and outcome of heart failure, and regulating myocardial energy metabolism has become an attractive treatment strategy for managing heart failure. Jiming formula (JMF), different from traditional Chinese medicine commonly used for heart protection, has been suggested to be effective in treating heart failure in experiments and clinical practice.

PURPOSE

This study integrated targeted metabolomics and transcriptomics to investigate the cardioprotective effects of JMF against myocardial infarction (MI) and the underlying molecular mechanism in mice.

METHODS

We first prepared a UHPLC-QTRAP-MS/MS method for analyzing JMF components. The cardioprotective effects of JMF in MI model mice were further identified using echocardiography, hematoxylin and eosin (HE) staining, Masson staining, Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, wheat germ agglutinin (WGA) staining and DHE staining. Differential gene expression in the hearts of the mice was detected using transcriptomics technology, and the cardiac metabolites were further quantified using LC‒ESI‒MS/MS. Network pharmacology was established to predict the cardioprotective components of JMF. Mitochondrial morphology and function in MI mice were evaluated using transmission electron microscopy and ATP assays. Finally, Western blotting and immunofluorescence were used to investigate the protective effects of JMF on the AMPK/SIRT 1/PGC-1 α signaling pathway.

RESULTS

A total of 191 and 40 components were identified in the JMF aqueous extract and rat plasma, respectively, indicating the quality of JMF. JMF improved survival rates and cardiac dysfunction in MI model mice in a quantitative manner and reduced adverse remodeling and mitochondrial damage. JMF protected cardiomyocytes from apoptosis and hypertrophy. Transcriptomic analysis revealed that JMF improved the mitochondrial tricarboxylic acid cycle (TCA cycle) in MI mice. Network pharmacology predicted that euodiae fructus may be the herb contributing the most to the effects of JMF. Targeted metabolomics analysis subsequently revealed that JMF treatment improved the substrate content in various pathways of glucose metabolism. JMF also improved poor metabolic remodeling in cardiomyocytes and enhanced glucose aerobic oxidation and ATP production. Enzyme assays revealed that JMF treatment increased the activity of key glycolytic enzymes and mitochondrial respiratory complexes I and IV. Furthermore, JMF activated the AMPK/SIRT1/PGC-1 α signaling pathway, resulting in the upregulation of GLUT4, PKM2, CPT1A and PPARα protein levels while reducing GLUT1 protein levels.

CONCLUSION

This research offers a novel perspective for treating MI using JMF. The underlying mechanism may involve the activation of the AMPK/SIRT1/PGC-1α signaling pathway and an increase in the aerobic respiration capacity of mitochondria. These findings provide valuable information regarding the pharmacological effects and mechanisms of JMF. In addition, this study provides a foundation for the application of euodiae fructus in the field of heart disease treatment.