This study aimed to uncover the mechanisms by which Agrimonia pilosa Ledeb. (APL) extract mitigates acute myocardial infarction (AMI) using a combination of network pharmacology and experimental validation. The main bioactive constituents of APL extract were identified through High-Performance Liquid Chromatography (HPLC). Potential targets shared between APL compounds and AMI were predicted via network pharmacology. An AMI model in mice was induced by subcutaneous isoproterenol (Iso) injection, followed by oral administration of APL extract. The study evaluated ECG patterns, cardiac injury markers, oxidative stress indicators, histopathology, PI3K/Akt signaling, and apoptosis-related proteins. Five active compounds were detected in APL extract. Functional enrichment suggested that cardioprotection by APL involves the PI3K/Akt pathway. APL treatment alleviated Iso-induced ST-segment elevation and tachycardia. It also reduced lactate dehydrogenase (LDH), creatine kinase (CK), CK-MB, malondialdehyde (MDA), and reactive oxygen species (ROS), while boosting superoxide dismutase (SOD) activity. Apoptosis of cardiomyocytes was markedly lowered after APL administration. Mechanistically, APL enhanced p-PI3K, p-Akt, and Bcl-2 expression and suppressed Bax, caspase-3, cleaved-caspase-3, and the Bax/Bcl-2 ratio in AMI mice. APL extract confers myocardial protection against Iso-induced AMI, likely through antioxidant and anti-apoptotic mechanisms mediated by the activation of the PI3K/Akt pathway.
