To investigate the toxicity of Esculentoside A on HK-2 cells (Human renal tubular epithelial cells).Methods：1)MTT assay was used to test the cell viability.2)The LDH in cells supernatant, SOD and MDA in the cells were detected. 3)Morphological changes of the cells were observed through inverted phase contrast microscope and electron microscopy. 4)Cell apoptosis was detected by Hochest33342-PI dyeing and flow cytometry.Results：1)Esculentoside A could inhibit nephrocyte viability dependently on time and dose (IC50 was 149.11μg·mL-1). 2)Esculentoside A could increase the activity of LDH in nephrocyte supernatant, reduce SOD activity and increase MDA content in nephrocytes. 3)Esculentoside A could cause damage of nephrocyte structure, cell apoptosis and necrosis.Conclusion：It showed that large dose of Esculentoside A (more than 100 μg·mL-1 ) has toxicity on HK-2 cells and the mechanisms of toxicity may include cellular oxidative damage and cell apoptosis.