Effect And Mechanism Of Allicin On Apoptosis Induced By H₂O₂ In HUVEC Cells

Objective Human Umbilical Vein Endothelial Cells (HUVEC) were treated with hydrogen peroxide (H2O2) to establish an oxidative stress apoptosis model, studying the effect of Allicin on apoptosis in HUVEC cells. Further more, the possible mechanism need to be explored at the molecular level and protein level.Methods HUVEC were treated with different concentrations (0.1mmol/L,0.5mmol/L and 1.Ommol/L) of H2O2 for 12h, apoptosis rate and secondary necrosis rate were measured by flow cytometry to establish a model of oxidative stress apoptosis. MTT method was used to detect the effects of Allicin on cell survival rate in oxidative stress apoptosis model under the different concentrations of allicin (1,10,20,40μg/mL) and the different action time (6,12, 24h). The contents of MDA, SOD and NO were measured in the medium. The expression of eNOS gene and iNOS gene were measured by RT-PCR method. The expression of apoptosis protein Caspase-3, PARP and Bax were measured by western-blot. So then the protective effects and the possible mechanism were to be explored by those methods.Results 0.5mmol/L H2O2 was suitable for the establishment of oxidative stress apoptosis model, because it could guarantee a certain rate of apoptosis, at the same time, the secondary necrosis rate was not significantly increased. Allicin could decrease the apoptosis rate induced by H2O2 in HUVEC cells. Allicin could also reduce the content of MDA, increase the content of SOD as well as the NO release in the medium. The expression of eNOS gene was increased by allicin significantly, the cutting of PARP was impaired, the expression of pro-Caspase-3 was increased, and the expression of Bax was decreased.Conclusion Allicin has powerful protective effects on HUVEC cells from apoptosis probably by increasing SOD, scavenging oxygen free radicals, preventing lipid peroxidation, increasing the expression of eNOS and NO release.