Objective：To investigate the expression changes of Notch3 in cisplatin-induced renal tubular epithelial cells（HK-2） injury and the potential regulation mechanism related to upstream microRNA. Methods：Based on HK-2 cells apoptosis model induced by cisplatin（20 μmol/L，24 h），Western blot and RT-qPCR were used to analyze the expression of Notch3 in HK-2 cells. Targetscan was used to search potential miRNAs that bind the 3’-UTR of Notch3. RT-qPCR was used to detect the expression of miR-192-5p in HK-2 cells after cisplatin treatment. Then，HK-2 cells transfected with negative control mimic（NC mimic） and miR-192-5p mimic respectively，and treated with or without cisplatin（20 μmol/L， 24 h），were divided into four groups：NC mimic group（NC）；NC mimic+cisplatin group（NC+Cis）；miR-192-5p mimic group（OE）；miR-192-5p mimic+cisplatin group（OE+Cis）. Western blot and RT-qPCR were used to demonstrate the effect of miR-192-5p on the expression of Notch3 after cisplatin treatment，while Western blot and flow cytometry were used to demonstrate the effect of miR-192-5p on apoptosis of HK-2 cells. Dual luciferase reporter assay was used to verified whether miR-192-5p targets Notch3. Results：Notch3 was up-regulated while miR-192-5p was down-regulated in HK-2 cells after cisplatin treatment compared with that in the normal controls. RT-qPCR showed that the expression of miR-192-5p in HK-2 cells was significantly increased after miR-192-5p mimic transfection. The overexpression of miR-192-5p alleviated cisplatin-induced apoptosis of HK-2 cells and negatively regulated the expression of Notch3. The dual luciferase reporter assay results verified that miR-192-5p directly binds with 3’-UTR of Notch3. Conclusion：miR-192-5p can attenuate the apoptosis of renal tubular epithelial cells induced by cisplatin through directly inhibiting the expression of Notch3.