Abstract: OBJECTIVE To explore the potential targets and mechanisms of Rhodiolae Crenulatae Radix et Rhizoma in regulating autophagy in granulosa cells based on network pharmacology and experimental verification. METHODS With the help of the database, the active components, effect targets and autophagy-related targets of Rhodiolae Crenulatae Radix et Rhizoma were obtained, and a component-target network diagram and a protein-protein interaction network diagram were established. Enrichment analysis of genes and signaling pathways was conducted on key targets and the active ingredients and core targets were docked to explore the possible mechanism of Rhodiolae Crenulatae Radix et Rhizoma in regulating autophagy in granulosa cells. Western blotting, RT-PCR, transmission electron microscopy and double-labeled adenovirus were used to detect the effect of important active ingredients of Rhodiolae Crenulatae Radix et Rhizoma on autophagy in granulosa cells. RESULTS There were 7 active ingredients of Rhodiolae Crenulatae Radix et Rhizoma obtained, with 441 targets, 796 autophagy-related targets, and there were 834 biological processes, 60 cellular components, 69 molecular functions, and 150 signaling pathways obtained by enrichment analysis. Molecular docking results showed that the active components of Rhodiolae Crenulatae Radix et Rhizoma and the core targets AKT1, ERBB2, PIK3CA, PIK3R1 could form a stable target binding effect. The experimental results showed that salidroside, an important active component of Rhodiolae Crenulatae Radix et Rhizoma, could promote the formation of autophagosomes and autophagolysosomes, and inhibit the activity of the Pl3K/AKT signaling pathway. CONCLUSION The results show that the regulation of autophagy in granulosa cells by Rhodiolae Crenulatae Radix et Rhizoma has the characteristics of multi-component-multi-target-multi-pathway, and its mechanism is related to the regulation of PI3K/AKT signaling pathway by salidroside.