Objective: To discuss the effect of Astragaloside Ⅳ (ASⅣ) on wound healing in rats with deep second degree burns by adjusting the Notch1/twist-related protein 1 (Twist1) signaling pathway. Methods: SD rats were randomly assigned into Con group, Model group, low-dose ASⅣ group (L-ASⅣ group), high-dose ASⅣ group (H-ASⅣ group), positive drug group (Shirun Shaoshang ointment group), and high-dose ASⅣ+Notch1 inhibitor DAPT group (DAPT group), with 15 rats per group. Except for the Con group, all other groups established deep second degree burn rat models. The healing rate and moisture content of burn wounds of rats in each group were detected. The levels of IL-8, IL-18, and IL-6 in serum of rats in each group were measured. The histopathological and apoptotic changes of burn wound tissues of rats in each group were measured. Moreover, the expression of epidermal growth factor (EGF), basic fibroblast growth factor (b-FGF), VEGF, platelet endothelial cell adhesion molecule CD31, and Notch1/Twist1 pathway proteins in burn wound tissues of rats in each group were measured. Results: Compared with the Con group, the Model group showed clear edema, structural integrity damage, and inflammatory cell infiltration in the burn wound tissue of rats, increased water content, IL-8, IL-18, IL-6, apoptosis rate, and the average fluorescence intensities of EGF and b-FGF, while decreased average optical densities of VEGF and CD31, and the expression of Notch1 and Twist1 proteins (P<0.05). Compared with the Model group, the L-ASⅣ group, H-ASⅣ group, and Shirun Shaoshang ointment group showed reduced tissue edema and inflammatory cell infiltration in burn wounds, restored structural integrity, increased wound healing rate, average fluorescence intensities of EGF and b-FGF, average optical densities of VEGF and CD31, expression of Notch1 and Twist1, and decreased water content, IL-8, IL-18, IL-6, and cell apoptosis rate (P<0.05). Compared with the H-ASⅣ group, the DAPT group showed increased tissue edema and inflammatory cell infiltration in burn wounds of rats, structural integrity damage, reduced wound healing rate, average fluorescence intensities of EGF and b-FGF, average optical densities of VEGF and CD31, expression of Notch1 and Twist1, and increased water content, IL-8, IL-18, IL-6, and cell apoptosis rate (P<0.05). Conclusion: ASⅣ may promote wound healing in rats with deep second degree burns by activating the Notch1/Twist1 signaling pathway.