Objective To explore the molecular mechanism of Smurf2's effect on hyperglycemia-induced mesangial cell proliferation and fibrosis by targeting EYA2. Methods Mouse mesangial cells (GMCs) were cultured, and sh-Smurf2 and sh-NC vectors were transfected into GMCs induced by high glucose. Cell proliferation in blank control group (NG), high glucose treatment group (HG), high glucose + sh-NC group (HG + sh-NC), and high glucose + sh-Smurf2 group (HG + sh-Smurf2) were detected by CCK-8 and EdU. The expressions of Fibronectin, Collagen I and α-SMA proteins were detected by Western blot. CO-IP assay was employed to detect the targeting interaction between Smurf2 and EYA2, and then sh-EYA2 and sh-NC vectors were used to transfect GMCs for backtracking experiments. Results Compared with NG group, cell proliferation and protein expression of Fibronectin, Collagen I and α-SMA in HG group were significantly increased. Compared with HG group, interference with Smurf2 inhibited cell proliferation and the expression of Fibronectin, Collagen I and α-SMA proteins. CO-IP results showed a targeted interaction between Smurf2 and EYA2. Compared with HG + sh-Smurf2 group, interference with EYA2 promoted cell proliferation and the expression of Fibronectin, Collagen I and α-SMA proteins. Conclusion Interference with Smurf2 can inhibit hyperproliferation and fibrosis of GMCs induced by high glucose by upregulating EYA2.