ETHNOPHARMACOLOGICAL RELEVANCE: Chronic heart failure (CHF) is characterized by complex pathogenesis involving various pathogenic mechanisms such as dysregulated autophagy, mitochondrial dysfunction, cardiomyocyte apoptosis, and chronic inflammation. Ershen Zhenwu Decoction (ESZWD) is a traditional Chinese herbal formulation. Several prospective, randomized, controlled clinical trials have indicated its clinical efficacy. However, whether ESZWD confers cardioprotective effects by regulating mitophagy and the associated molecular pathways remains elusive.
AIM OF THE STUDY: This study aimed to investigate whether ESZWD ameliorates CHF by modulating PTEN-induced putative kinase 1(PINK1)/Parkin pathway mediated mitophagy.
METHODS: UHPLC-Q-TOF MS was performed to identify the chemical compounds in ESZWD. Network pharmacology, molecular docking and dynamics were predict possible therapeutic targets. A rat model of CHF with heart-kidney yang deficiency syndrome was created for in vivo investigations via thyroidectomy and doxorubicin induction, followed by treatment with ESZWD. Furthermore, the impact of ESZWD was determined by monitoring cardiac function, serum biomarkers (ATP, ADP, NT-proBNP), myocardial histopathology, mitochondrial ultrastructure, and ROS levels. In addition, the expression of mitophagy-related proteins (PINK1, Parkin, LC3II/I, Beclin-1 and p62) in myocardial tissues was assessed via Western blotting. In vitro, DOX-stimulated AC16 cardiomyocyte injury model cells were exposed to ESZWD or the autophagy suppressor 3-methyladenine (3-MA). Then, the expression of PINK1, Parkin, LC3II/I, Beclin1 and p62 was assessed by PCR and Western blotting. Similarly, the cellular localization and expression of PINK1 and Parkin were confirmed by immunofluorescence staining.
RESULTS: The data identified 785 compounds in ESZWD, with saponins, terpenes, alkaloids, phenolic acids and tanshinones being particularly abundant. Network pharmacology analysis revealed that ESZWD's therapeutic benefits via the PINK1/Parkin signaling pathway to regulate mitophagy, decreasing CHF onset and progression. Molecular dynamics simulations demonstrated persistent interactions between the active chemical 8-gingerol and the PINK1 protein. In vivo, ESZWD therapy improved cardiac function in CHF rats and mitophagy-related proteins, indicating its ability to inhibit mitophagy. In vitro experiments confirmed that ESZWD significantly reduced autophagy-related proteins, with effects consistent with those of the autophagy inhibitor 3-MA, validating that ESZWD mitigates pathological mitophagy.
CONCLUSIONS: This study preliminarily revealed that ESZWD may alleviates CHF by modulating the PINK1/Parkin signaling pathway to inhibit mitophagy.
