Abstract:To investigate the DNA damage from oxidative stress in human umbilical vein endothelial cells (hUVECs) induced by cholesterol, the γH2AX foci were detected by immunofluorescence following cholesterol treatments at 125 mg/L, 25 mg/L, 50 mg/L for 12 hours. The results showed that formation of γH2AX foci was increased as measured by the numbers fluorescence intensity. Western bolt and flow cytometry (FCM) showed that the level of γH2AX and reactive oxygen species (ROS) increased with cholesterol doses. Using comet assay to detect DNA lesions, we found higher percentage of tailing cells as the cholesterol concentration increased. Preexposed to 10 mmol/L Nacetyl6cysteine(NAC) for 1 hour, the hUVECs treated with 50 mg/L cholesterol showed significantly reduced γH2AX foci and decreased ROS. The results indicated that cholesterol induced DNA damage in hUVECs by elevated ROS levels.
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