| Obiective:The aim of this study was to explore whether human umbilical vein endothelial cell with pretreatment of PUE attenuated the effect of IRON.and to Futher exploring the mechanism how eNOS works in the process that puerarin protected against human umbilical vein endothelial cell from iron damage.Method:This study established an IRON injury model by selecting well-established HUVECs.The cultured HUVECs endothelial cells were randomly divided into 8 groups,each group repeated three times:(1)Control group;(2)IRON group;(3)20μM PUE+IRON group;(4)40μM PUE+IRON group;(5)80μM PUE+IRON group;(6)160μM PUE+IRON group;(7)320μM PUE+IRON group;(8)L-Arg+IRON group.Firstly,the cell viability of each experimental group was measured by the MTS assay,The release of LDH was measured by spectrophotometer.Flow cytometry was used to detect the level of reactive oxygen species(ROS)and the degree of apoptosis.Mitochondrial permeability transition pore(mPTP)was opened by spectrophotometry,and Western blotting was used to detect the expression of eNOS protein.Results:1.Compared with IRON group,HUVECs with pretreatment of PUE all significantly increased cell viability and decreased intracellular LDH release(p<0.01).There was no statistical significance between 160μM PUE group and 320μM PUE group.the best effect was 160μM PUE.2.Compared with the IRON group,the PUE pretreatment group and the L-Arg pretreatment group showed a decrease in ROS levels of HUVECs(p<0.05),a significant decrease in apoptosis(p<0.01),and up-regulation of eNOS protein expression(p<0.01).3.Compared with IRON group,the opening of mPTP in HUVECs cells was significantly inhibited in PUE pretreatment group and L-Arg pretreatment group.Conclusion:PUE pretreatment protected HUVECs cells from IRON damage.Its mechanism was possibly associated with up-regulation of eNOS protein expression. |
