Effect Of Adenovirus-mediated GPX Gene Transfection On Oxidative Stress And Apoptosis Of Endothelial Cells Induced By Cigarette Smoking Extract

Objective: To elucidate the effect of CSE on oxidative injury and apoptosis of endothelial cells, and whether transfection of Adenovirus-mediated GPX to endothelial cells could partly inhibit the effectMethods: The endothelial cells(ECV304) were cultured in vitro and divided into blank group, CSE group, LacZ group and GPX group. Apoptosis index (AI) of ECV304 were measured by TUNEL. Intracellular reactive oxygen species(ROS) were detected by H2DCFDA staining and laser confocal microscopy.Results:1. transfection efficiency of 70MOI Adenovirus-mediated LacZ was (93.3±2.9)%;2. The AI of 10%CSE interfered ECV304 group was higher than the 0%CSE and 5%CSE interfered groups (p<0.05). After 10%CSE interfered ECV304 for 24 hours, the AI was higher than the 0h,6h and 12h groups(p<0.05);3. The AI and the levels of Intracellular ROS in the 10%CSE group were higher than the blank group (p<0.05);4. Adenovirus-mediated GPX gene transfection could decrease the levels of Intracellular ROS and AI of ECV304 induced by CSE (p<0.05); 5. There was a positive correlation between the levels of Intracellular ROS and the AI of ECV304(n=5, r=0.775,p<0.01).Conclusion:1. CSE induces oxidative stress and apoptosis of endothelial cells.2. Transfection of Adenovirus-mediated GPX gene to endothelial cells could partly inhibit the oxidative injury and apoptosis.3. CSE induces apoptosis of endothelial cells may be related to oxidative stress.