Changes Of Sirturin-3Protein Expression Levels Under The Stress Of High Concentration Of Glucose And Oxidized Low Density Lipoprotein

Objective:1. To establish a stress model of high glucose by dealing HUAEC with culture medium that contains a series of different concentration D-glucose.2. To establish a stress model of lipoprotein by dealing HUAEC with culture medium that contains ox-LDL.3. At the same time this investigation try to explore the effects of full-length sirt3on the anti-appoptosis mechanism of HUAEC under stress.4. Try to investigate the relationship between full-length sirt3and atherosclerosis.5. Try to shed light on the subcelluar localization of full-length sirt3.Methods:1. The HUAECs, as a kindly gift of professor Zhao yu-xia,were cultured carefully before divided randomly into the following groups:normal control group,11mmol/L D-glucose group,22mmol/L D-glucose group,44mmol/L D-glucose group,and40ug/ml ox-LDL group。2. Those cells were harvested after being treated for48hours.3. The expression of full-length sirt3at protein level was examined by means of Western-blot. Analyses were performed using the statistical software package SPSS19.0. Results:1. After being dealt with gradient concentration D-glucose for48hours, the endothelial cells grew slower as the concentration of D-glucose get higher, evidenced by smaller cell density and more dead cells. Time limited,we did not meter these changes with precise methods.2. Full-length sirt3protein was found in the cytoplasm of normal control group.3. Full-length sirt3protein was found in the cytoplasm of each and every high-glucose groups.4. There is no obvious differences among tublin expression levels of all the groups (P>0.05)5. Compared with the normal control group,each and every group that was dealt with high glucose got a up-regulated protein expression level of full-length sirt3in cytoplasm, and the differences were obvious (p<0.05).And the difference among different D-glucose concentration groups are obvious (P<0.05),besides we found a elevated trend of protein expression level of full-length sirt3in cytoplasm as the concentration of D-glucose got higher.6. After being dealt with ox-LDL for48hours, the endothelial cells grew slower as the concentration of D-glucose get higher, evidenced by smaller cell density and more dead cells. Time limited,we did not meter these changes with precise methods.7. Full-length sirt3protein was found in the cytoplasm of HUAECs dealt with ox-LDL group.8. Compared with the normal control group,ox-LDL group got a up-regulated protein expression level of full-length sirt3in cytoplasm, and the differences was obvious (p<0.05)Conclusion:1. Full length sirt3protein presents in the cytoplasm of normal HUAECs.2. The protein expression level of full-length sirt3in cytoplasm is up-regulated under transient high glucose stimulation, and these expression levels were obviously elevated as the concentration of D-glucose get higher, suggesting that sirt3protein is likely to take part in the mechanism against the damage caused by hight concentration glucose in endothelial cells.3. The protein expression level of full-length sirt3in cytoplasm is up-regulated under transient ox-LDL stimulation, suggesting that sirt3protein is likely to take part in the mechanism against the damage caused by ox-LDL in endothelial cells.4. sirt3is likely to play a role in the mechanism of anti-stress mechanism in endothelial cells,and the stress is probably nonspecific.5. We build models of high glucose and ox-LDL transient stimulation on endothelial cells that are similar with in vivo, which could be helpful for further studies about full-length sirt3and endothelial damage caused by glucose and ox-LDL,and the relationship between full-length sirt3and atherosclerosis.