To determine therapeutic effect of Qishen Huoxue Granule (QHG) on the myocardial injury in sepsis and whether it is through the inhibition of excessive autophagy by using network pharmacological analysis and in vitro.120 SPF male Wistar rats were divided into 6 groups. Firstly, the function of the heart were evaluated through echocardiography, and pathological changes of myocardial tissue was observed by Hematoxylin-eosin (H&E) and Transmission electron microscopy (TEM). Subsequently, enzyme-linked immunosorbent assay (ELISA) was employed to measure the levels of myocardial injury markers (cTnT and BNP) and inflammatory cytokines (TNF-α and IL-1β). Finally, the expression of proteins associated with the MasR/PI3K-AKT-mTOR pathway and autophagy-related proteins was evaluated using immunohistochemistry, immunofluorescence, RT-qPCR, and Western blot analysis.The results demonstrated that QHG significantly improved systolic function in rats, reduced levels of myocardial injury markers and inflammatory cytokines, and downregulated autophagy-related genes (ATG5, LC3, and Beclin1). However, when QHG was combined with inhibitors of the MasR/PI3K-AKT-mTOR pathway, the mRNA and protein expression levels of pathway components were reduced. Concurrently, the mRNA and protein expressions of autophagy indicators were upregulated, counteracting the inhibitory effect of QHG on excessive autophagy.QHG ameliorated the myocardial injury in sepsis rats by inhibiting the excessive autophagy in myocardial cells via activating the MasR/PI3K-AKT-mTOR pathway.