Effect Of Simvastatin And Probucol On Endothelial Cell Apoptosis Mediated By Tumor Necrosis Factor Alpha

ObjectiveThe purposes of this study were to determine endothelial cell apoptosis induced by tumor necrosis alpha,and preliminary study effect of cytokines such as TNF-a in the pathogenesis of heart failure,as well as the associate between cytokines and cell apoptosis.To investigate the effect of simvastatin and probucol on endothelial cell apoptosis mediated by tumor necrosis alpha.MethodHuman umbilical vein endothelial cells(HUVEC) were cultured in vitro.The cells for experiments were in logarithmic growth phase.1.In accordance with the concentration of TNF-αinterfere with the cells divided into four groups:control group, 1ng/ml TNF-αintervention group, lOng/ml TNF-a intervention group,25ng/ml TNF-αintervention group. After 12h cells in each group intervention, using flow cytometry to detect cell apoptosis rate.2.According to the results of the previous step, selecting 10ng/mlTNF-αrespectively to interfere with cell Oh,12h,24h, 48h,and then using flow cytometry to detect cell apoptosis rate. 3.Comprehensive experimental results the first two steps, lOng/ml TNF-a with different concentrations of simvastatin co-intervention in cells, which was divided into four groups:10ng/mlTNF-a+Oμmol/l simvastatin group, 10ng/ml TNF-α+0.01μmol/l simvastatin group, 10ng/mlTNF-α+0.1μmol/l simvastatin group, 10ng/mlTNF-α+1.0μmol/l simvastatin group,12h intervention in each group and then using flow cytometry to detect cell apoptosis rate.4.Comprehensive experimental results the first two steps, lOng/ml TNF-a with different concentrations of probucol co-intervention in cells, which was divided into four groups:lOng/ml TNF-α+0μmol/l probucol group,10ng/ml TNF-α+20μmol/l probucol group,10ng/mlTNF-α+40μmol/l probucol group,10ng/ml TNF-α+80μmol/l simvastatin group,12h intervention in each group and then using flow cytometry to detect cell apoptosis rate.Result(1)Different concentrations of TNF-a, after 12h intervening endothelial cells,apoptosis detected by flow cytometry were:control group 0.81±0.25%,the results of the intervention groups were 2.35±0.04%,4.54±0.32% and 6.17±0.28%,respectively,there were significant differences amony of groups.(2) 10ng/mlTNF-a intervention endothelial cells after different time,apoptosis detected by flow cytometry were:control group 0.87±0.35%,the results of the intervention groups were 4.53±0.32%,7.45±1.97% and 10.92±1.27% respectively, there were significant differences among of groups.(3)10ng/mlTNF-α with different concentrations of simvastatin co-intervention in cells after 12h, apoptosis detected by flow cytometry were:control group 4.46±0.53%,the results of the intervention group were 1.38±0.12%, 2.89±0.27% and 3.65±0.08%, respectively,there was a significant difference between each intervention group and control group. (4)10ng/ml TNF-a with different concentrations of probucol co-intervention in cells after 12h, apoptosis detected by flow cytometry were:control group 4.60±0.64%,the results of the intervention group were 2.61±0.18%,2.21±0.22%and 1.28±0.34%,respectively, there was a significant difference between each intervention groups and control group.Conclusion(1)TNF-a can induce apoptosis of endothelial cells by dose-dependent and time dependent.(2) Simvastatin can inhibit TNF-a-induced apoptosis of endothelial cells, but no concentration-dependent. With the increasing concentrations of simvastatin, induced the cell apoptosis induced increase first, but decrease then.(3)Probucol can inhibit TNF-a-induced apoptosis of endothelial cells, but no concentration-dependent, with the increase of the concentration of probucol,there was no difference in anti-apoptosis enhancement.