AIM: To explore the protective mechanism of Mongolian medicine Gaoyou on renal function through network pharmacology molecular docking and animal experiments. METHODS: The effective active components and targets of Gao you acting on acute kidney injury (AKI) and the related targets of AKI disease were obtained by using the relevant database of traditional Chinese medicine. The targets and components were mutually constructed to act on the network, and the signal pathways of drugs and diseases were enriched and analyzed. And the molecular docking validation study was carried out on the pharmacodynamic components with high moderate value in the network and the core action target. On this basis, after establishing the rat model of acute renal injury and giving Gaoyou intervention, the contents of creatinine (CR), urea nitrogen (BUN) and tumor necrosis factor-A (TNF-α), Kidney damage molecule-1 (Kim-1), cystatin C (Cys-C), neutrophil gelatinase related lipid transporter (NGAL) in serum of rats were detected, and renal tissue sections were observed with HE staining to verify the protective effect of Mongolian medicine Gaoyou on renal function of rats with acute renal injury. The predicted signal pathway was verified by Western blot. RESULTS: 40 key targets were obtained by network pharmacology and molecular docking analysis. KEGG and go enrichment analysis were carried out on the key targets. ALB, AKT1, VEGFA, IL-6, CASP3 and other key targets were predicted, and cancer signal pathway, TNF signal pathway, PI3K/AKT signal pathway and other key pathways were predicted. The results of molecular docking showed that the main active components of Gaoyou had good binding activity with related targets. The results of animal experiments showed that Gao you had a good protective effect on renal function, and Cr and BUN were significantly lower than those in the model group (P<0.05). The content of TNF-α decreased significantly (P<0.05). Kim-1, Cys-C and NGAL decreased significantly (P<0.05). HE staining showed that Gao can obviously improve the pathological state of acute renal damage. The expression of AKT1, p-AKT1 and CASP3 in renal tissue of rats with acute renal injury increased (P<0.05). Compared with the model group, the expression of AKT1, p-AKT1 and CASP3 in renal tissue of rats in Gaoyou group decreased significantly (P<0.05). CONCLUSION: Gaoyou can protect the renal function of rats with renal injury through multiple pathways such as anti-inflammatory, antioxidant, antithrombotic, anti apoptotic and so on through the characteristics of multi-component, multi target. Intervention of PKA3/AKT signal pathway and apoptosis factor may be one of its main mechanisms.