Objective To investigate the effect of aucubin on the inflammatory injury of renal tubular epithelial cells induced by high glucose and its molecular mechanism. Methods Renal tubular epithelial cells HK-2 were divided into control (Con) group, high glucose (HG) group, aucubin low, medium and high concentration (HG+AU-L, M, H) groups, and HG+miR-NC Group, HG+miR-374 group, HG+AU+anti-miR-NC group, HG+AU+anti miR-374 group. Enzyme linked immunosorbent assay (ELISA) was used to detect IL-6 and TNF-α levels. The flow cytometry was used to detect cell apoptosis. The real time fluorescent quantitative PCR (RT-qPCR) was used to detect miR-374 expression level. Results The levels of IL-6 and TNF-α in renal tubular epithelial cells induced by high glucose were increased, the rate of apoptosis was increased, and the expression level of miR-374 was decreased (P＜0.05). After treatment with different concentrations of aucubin, the levels of IL-6 and TNF-α were decreased, the rate of apoptosis was decreased, and the expression level of miR-374 was increased, in a concentration-dependent manner (P＜0.05). After the overexpression of miR-374, the levels of IL-6 and TNF-α in renal tubular epithelial cells induced by high glucose were decreased, and the apoptosis rate was decreased (P＜0.05). Inhibition of miR-374 expression reversed the effect of aucubin on the inflammatory injury of renal tubular epithelial cells induced by high glucose. Conclusion Aucubin inhibits the inflammatory injury of renal tubular epithelial cells induced by high glucose by regulating the expression of miR-374.