Objective：To investigate the expression changes of Notch3 in cisplatin-induced renal tubular cell injury and the potential upstream regulation mechanism of its related microRNA. Methods：Human renal tubular epithelial cells (HK-2 cells) were cultured in vitro and transfected with miR-192-5p mimic (miR-192-5p mimic) and negative control mimic (NC mimic), respectively. Then, they were divided into blank control group and cisplatin treatment group (20μmol/L). After 24 hours, The mRNA expression levels of miR-192-5p and Notch3 were quantified by real-time quantitative PCR and the protein expression levels of Notch3 and apoptosis-related protein cleaved-caspase3 were detected by Western blotting. The apoptosis rate of cells was evaluated by flow cytometry. The reporter plasmids containing the 3'non-coding region (3'-UTR) of Notch3 was constructed and the luciferase activity of target gene Notch3 were detected by dual luciferase report assay. Results：After 24 hours of cisplatin treatment, compared with the blank control group, the expression of miR-192-5p was reduced, while the mRNA and protein expression of Notch3 increased, and the expression of apoptosis-related protein clear-caspase3 protein and apoptosis rate increased in HK-2 cells (all p <0.05). Then, compared with NC mimic group treated with cisplatin, overexpression of miR-192-5p markedly decreased the mRNA and protein expression of Notch3, which was predicted to be the target gene of miR-192-5p. Meanwhile, the protein expression of apoptosis-related protein cleaved-caspase3 and apoptosis rate was significantly lower than that of cisplatin group (all p<0.05). The dual luciferase reporter assay showed that miR-192-5p+pmiR-Notch3 3’-UTR-WT co-transfection group had lower Notch3 luciferase activity than miR-192-5p+pmiR-Notch3 3’-UTR-Mut group, demonstrating the direct interaction between miR-192-5p and Notch3 (P<0.05). Conclusions: miR-192-5p can attenuate the apoptosis of renal tubular epithelial cells induced by cisplatin through directly inhibiting the expression of Notch3.