Objective: To investigate the role of circAff4 in high glucose-cultured podocytes in vitro and its relationship with Notch pathway and nephrin. Methods: After culturing podocytes in 30mmol?L-1 high glucose for 0, 12, 24, 48, and 72 h, the levels of circAff4 were measured using PCR. Following the overexpression of circAff4 via an overexpression vector, the levels of Notch1, Hes1, and nephrin were assessed using immunocytochemistry, Western blot, and PCR methods. Results: High glucose cultivation of podocytes could inhibit circAff4 expression (P<0.05 or P<0.01). After adding the circAff4 overexpression vector to high glucose-cultured podocytes to increase circAff4 expression, the expression of Notch1 and Hes1 significantly increased, while the expression of nephrin significantly decreased (P<0.01). Conclusion: In high glucose-cultured podocytes, circAff4 may participate in podocyte injury through the Notch pathway and nephrin.