AIM: To observe the effect and mechanism of cuttlebone extract regulating vascular endothelial growth factor (VEGF)/phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway on refractory wounds in rats. METHODS: Cuttlebone extract(CE) was obtained by water extraction of cuttlebone. Fifty SD rats were randomly divided into negative Control group, Model group, Cuttlebone extract low-dose (CE(L)) group, Cuttlebone extract high-dose (CE(H)) group, and cuttlebone extract high-dose+inhibitor (CE(H)+LY294002) group. After the refractory wound model was successfully established, 0.02% furacillin solution or cuttlebone extract solution were applied to the wound area of rats in each group, and the treatment was administered once a day. After 14 days of treatment for refractory wounds, the changes in wound healing, angiogenesis, inflammation and expression of related regulatory proteins were quantitatively analyzed by measuring skin ulcer wound area, pathological sections, immunofluorescence staining, Elisa, Western blot, RT-qPCR and other methods. RESULTS: Compared with Model group, CE(L) and CE(H) groups can increase the number of epithelial cells and collagen, and promote the healing of refractory wound in rats. Serum VEGF, skin tissue microvascular density, P-PI3K, P-AKT, VEGF protein expression and mRNA expression levels of PI3K, Akt, VEGF and eNOS were increased (P<0.05), while serum TNF-α and IL-6 levels were decreased (P<0.05). LY294002 could partially reverse the repairing effect of high dose cuttlebone extract on refractory wound (P<0.05). CONCLUSION: Cuttlebone extract can regulate the VEGF/PI3K/AKT signaling pathway, inhibit the inflammatory response of refractory wounds in rats, induce angiogenesis and promote wound healing.