This research aimed to elucidate the mechanisms through which the Zuogui Jiangtang Shuxin formula (ZGJTSXF) alleviates diabetic cardiomyopathy (DCM) using a combined approach of serum pharmacochemistry, network pharmacology, and experimental validation.Following oral administration of ZGJTSXF in rats, serum compounds were analyzed using UPLC-Q-Exactive-Orbitrap-MS. A network linking bioactive components of ZGJTSXF to DCM-associated targets was established with Cytoscape. Functional enrichment analyses, including Gene Ontology and KEGG pathway assessments, were performed to identify critical molecular targets and signaling pathways. DCM model mice were treated with ZGJTSXF, and the predicted key pathways were validated through qPCR and Western blot assays.Analysis identified 78 serum compounds, including flavonoids, peptides, nucleosides, organic acids, phenylpropanoids, alkaloids, phenanthrenequinones, iridoids, phenols, and saponins. Network pharmacology indicated that ZGJTSXF may influence targets such as ALB, TNF, AKT1, GAPDH, VEGFA, EGFR, SRC, CASP3, MAPK3, JUN, and modulate the PI3K/AKT pathway in DCM. Administration of ZGJTSXF improved glucose metabolism, cardiac function, and myocardial tissue structure in DCM mice. Importantly, ZGJTSXF reduced cardiomyocyte apoptosis, which correlated with activation of the PI3K/AKT signaling pathway and increased levels of anti-apoptotic proteins Bcl-2 and Bcl-xL.These findings provide evidence that ZGJTSXF exerts cardioprotective effects in DCM via PI3K/AKT pathway activation and inhibition of apoptosis. The study also identifies the potential bioactive constituents responsible, offering a basis for future development of ZGJTSXF-based therapies for diabetic cardiomyopathy.
