This study preliminarily investigated the potential mechanisms of the Huoxue Tongluo prescription(HXTLP) in treating spinal cord injury(SCI) through a combination of network pharmacology, molecular docking technology, and in vivo experiment verification. TCMSP database and Swiss Target Prediction platform were utilized to select the active ingredients and targets in HXTLP with therapeutic properties for SCI, and the "active ingredients - targets" network was construted.SCI related targets were searched in OMIM and GeneCards, and the protein interaction network (PPI) of the common targets of HXTLP and SCI was established based on the STRING database. The Metascape database was used for KEGG pathway enrichment and GO analysis of the common targets. Molecular docking of active ingredients and key targets was performed through Autodock1.5.7 software and the results were visualized by Pymol software. Finally, the effect of HXTLP on SCI was verified by animal experiments.The results indicated that A total of 184 intersection targets were obtained, and the key targets were AKT1, STAT3, HSP90AA1, PIK3CA, PIK3R1, HRAS, MAPK1, EGFR, etc. Molecular docking results showed a strong binding ability between the core active components and key targets. Animal experimental results showed that HXTLP group had significant therapeutic potential for SCI compared with model group. Behavioral scores and footprint analysis showed improved hind limb motor function, and histological examination showed more complete structure and morphology of the injured area.WB result revealed that HXTLP can effectively inhibit the key target protein HSP90AA1 and the expression of P-STAT3, promoted the expression of P-AKT1. This study verifies that HXTLP has the characteristics of multi-component, multi-target and multi-pathway synergistic effect in the treatment of SCI, and provides experimental theoretical basis for clinical medication and further research of SCI.