Abstract: Objective To investigate the effect of N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP) on podocyte injury induced by high glucose and its possible mechanisms. Methods The conditionally immortalized mouse podocytes were treated by high glucose with or without AcsDKP. The expression of nephrin and podocyte cytoskeleton was detected by immunofluorescence. Western blotting was used to detect the expression of fibronectin and α-SMA protein in podocytes. Flow cytometry was used to measure podocyte apoptosis. Results High glucose inhibited the expression of nephrin, and AcSDKP restored the expression of nephrin. High glucose promoted the expression of fibronectin and α-SMA, and AcSDKP inhibited the expression of fibronectin and α-SMA. High glucose induced the reorganization of podocyte cytoskeleton and decreased the podocyte adhesion, and AcSDKP rescued the podocyte cytoskeleton and podocyte adhesion. High glucose induced podocyte apoptosis, and AcSDKP inhibited podocyte apoptosis. Conclusions AcSDKP alleviated podocyte injury induced by high glucose via rescuing epithelial-mesenchymal transition in podocytes, making cytoskeleton stable and inhibiting podocyte apoptosis.