Mufangji decoction ameliorates myocardial hypertrophy by regulating apoptosis via the PI3K/AKT pathway

Abstract

Mufangji decoction(MFJD), a classical traditional Chinese medicine formula, has been clinically used for cardiovascular disorders such as heart failure. However, its use in treating cardiac hypertrophy is relatively limited, and its specific mechanisms of action remain unclear. This study aims to investigate the active components and underlying mechanisms of MFJD in treating myocardial hypertrophy using network pharmacology, molecular docking, and in vitro experiments. We screened active components of MFJD from the TCMSP database and supplemented mineral components from Gypsum Fibrosum based on the literature. We predicted potential targets using PubChem and Swiss Target Prediction, retrieved myocardial hypertrophy-related targets from OMIM, GeneCards, and DrugBank, and identified common targets using Venn analysis. We then imported the intersecting targets into the STRING database to construct a PPI network, performed GO and KEGG enrichment analyses in R, and established a Disease-target-component-pathway network in Cytoscape. Subsequently, we performed molecular docking using AutoDockTools and PyMOL. Based on these findings, we established an isoproterenol-induced HL-1 cell model to simulate in vitro myocardial hypertrophy and divided cells into control, model, and MFJD-treated groups. We evaluated the therapeutic effects and mechanisms of MFJD using CCK-8, Western Blot, and Annexin V-FITC/PI apoptosis assays. In addition, we performed a rescue experiment with the PI3K inhibitor LY294002 to verify pathway specificity. Network pharmacology identified 33 active components and 93 common targets associated with myocardial hypertrophy, with the PI3K/AKT pathway being significantly enriched. In vitro experiments demonstrated that MFJD reduced ISO-induced increases in cell area and BNP mRNA expression, decreased apoptosis rates, and upregulated the expression of p-AKT/AKT, PI3K/GAPDH, and Bcl-2/Bax proteins. Notably, MFJD exhibited a non-monotonic concentration–response pattern, with the 100 μg/ml group showing the most pronounced phenotypic effects. Furthermore, LY294002 partially reversed MFJD-induced restoration of PI3K/AKT signaling and the Bcl-2/Bax balance, supporting the involvement of the PI3K/AKT pathway in MFJD’s protective effects. MFJD alleviates ISO-induced myocardial hypertrophy and apoptosis, and these effects are associated with activation of the PI3K/AKT signaling pathway. The findings provide mechanistic insights into the potential protective effects of MFJD against myocardial hypertrophy and support further in vivo and clinical investigations.