The Role And Mechanisms Of BYHWD On Hydrogen Peroxide-induced Apoptosis In Human Umbilical Vein Endothelial Cell

Background:Vascular endothelial cell injury is an important factor which aggravates brain damage and affects the recovery of neurological function after cerebral infarction, the mechanisms of which due to the large production of oxygen free radicals after cerebral ischemia and reperfusion injury. BYHWD is a traditional Chinese herbal medicine which protects nerve cell function shown in many studies, however, the roles and mechanisms on endothelial cell injury after cerebral infarction has not been fully explained.Objective:In this study, we establish H2O2-induced human umbilical vein endothelial cells (HUVECs) injury in vitro, as a oxidative stress injury model after schemia-reperfusion injury, by which investigate the protective effect of BYHWD and its possible mechanisms.Methods:To study the protective effect on HUVECs, we detect cell viability by MTT, the number of apoptotic cells by Hoechst33342/PI apoptosis protein Caspase-3expression by Western blot. To investigate the effect on mitochondria, we observe the mitochondrial morphology by TEM,detect mitochondrial membrane potential (MMP) by JC-1, To study the mechanism against oxidative stress, we detecting intracellular ROS by DCFH-DA, superoxide dismutase (SOD) and malondialdehyde (MDA).Results:Compared with H2O2group, BYHWD group (1mg/ml) was shown no significant difference in percentage of apoptotic cells (p>0.05), while BYHWD groups (15,30mg/ml) were significantly different (p<0.01); compared with H2O2group, BYHWD group (30mg/ml) was significantly decreased (p<0.01) in caspase-3expression; BYHWD group (30mg/ml) significantly improve mitochondrial structure, increase the MMP (p<0.01), reduce ROS production, increased SOD and decrease MDA (p<0.01).Conclusion:In conclusion, BYHWD significantly reduced the H2O2-induced apoptosis in HUVECs, reversed the decline in MMP, improved the mitochondria morphology, reduce the generation of ROS, increased SOD activity and decrease MDA production. The result indicated that BYHWD prevented H2O2-induced apoptosis of HUVECs through regulating reactive oxygen species mediated mitochondrial dysfunction pathway.