Abstract: To explore the expression of HDPD1 gene in EA.hy926 cell apoptosis induced by high glucose and hight cholesterol. Methods: cells were incubated with high or low concentrations of glucose (25 mmol/L ) and cholestrol (0.8 , 1.6 ,3.2 mmol/L) for different times. We detected the cell activity by CCK8 Kit, observed the cell apoptosis by FACS (Flow Cytometer), and measured the expression of HSPD1 gene by RT-PCR and QPCR. Results: Cholesterol (0.8 , 1.6 , 3.2 mmol/L) together with high glucose strikingly decresed cell activity and in a dose-dependent manner, the same as the cholesterol (1.6 , 3.2 mmol/L) together with low glucose, but not for the cholestrol (0.8 mmol/L) with low glucose. The cell activity also decreased in a time dependent manner when treated with low or high glucose + 0.8 mmol/L. Most of cells treated with hight glucose and hight cholesterol (high glucose + 0.8 mmol/L cholesterol) underwent apoptosis compared with the low glucose and high cholesterol (high glucose + 0.8 mmol/L cholesterol). Meanwhile, the expression of HSPD1 gene were dramatically induced by high glucose and high cholesterol.