Protection Effect Of Shuxuening Human Umbilical Vein Endothelian Cells Injury Induced By Oxidize Low Density Lipoprotein

OBJECTIVEThis experiment through In vitro foster human umbilical vein endothelial cells (HUVEC), including primary culture and subculture, and identified on the cultured cells. By oxidized low density lipoprotein (ox-LDL) on cultured HUVEC caused lipid peroxidation. Observed Shuxuening's Protective effects on endothelial cells and explore its possible mechanism.METHODSTake Fresh umbilical cord vein endothelial cells in vitro primary and passaged cultured. And identified the cultured cells Established model of lipid peroxidation. Divided by the different dosing: control group, ox-LDL injury group, shuxuening plus ox-LDL group( the cells were pretreated by shuxuening Injection for 24h,then add the ox-LDL)and Normal saline group (the cells were pretreated by Normal saline for 24h,then add the ox-LDL). Using methyl thiazolyl tetrazolium (MTT) to detect cell's Relative number and relative viability; Using Western Blot,ELISA observed ox-LDL stimulation in 0h, 3h, 12h, 24h, protein levels of the endothelial cell ICAM-1and MCP-1. By Technique of protein blotting (Western Blot) observed stimulation of 12h, ox-LDL on human umbilical vein endothelial cells ICAM-1, MCP-1's protein levels. And the injection with Shuxuening changes of the factors. On the basis of the results, to analysis the protection effects of Shuxuening injection impact on vascular endothelial cells. and to explore its possible mechanism. All quantitative data were analyzed by SPSS13.0 software.RESULTS(1)MTT detected relative number of endothelial cells and its relative activity: ox-LDL incubated with HUVEC after 24h reduced the activity of the HUVEC,the OD value compared with the control group decreased significantly (P<0.01); the OD value of the Shuxuening injection group was significantly higher than ox-LDL group (p <0.01).(2)Shuxuening impact on the morphology of HUVEC that processed by ox-LDL. Observed under an inverted microscope: Normal control group HUVEC showed a single layer, its borders clear cytoplasm rich, not each other, into a typicalcobblestone mosaic arrangement, after incubated with In ox-LDL, the cells shrinking, widened intercellular space, cell membrane shrinkage, showing off some cells, the cell boundary is not clear. In the Shuxuening injection group, cell morphology is normal, but the gap narrowed.(3)Shuxuening Impact on the expression ox-LDL induced HUVEC ICAM-1 and MCP-1 protein. Enzyme-linked immunosorbent assay (ELISA) detection of ox-LDL-induced HUVEC of ICAM-1, MCP-1 protein expression increased. After given the Shuxuening injection, the protein expression reduced. Analysis showed that after the stimulation of ox-LDL at each time point both ICAM-1 and MCP-1 expression increased .There was a significant difference (P <0.01). 3h, 6h, 12h, 24h each time point compared with the control group. ICAM-1 levels were Respectively increased by 28.9%, 44.7%, 89.9%, 81.9% MCP-1 levels were increased by 30.7%, 57.9%, 130%, 120%. Among the most significant increase is 12h group. Compared with ox-LDL injured group, after given injection Shuxuening ICAM-1, MCP-1 expression decreased at all time points ICAM-1 at 3h, 6h, 12h, 24h, respectively at each time point fell 7.13%, 13.2%, 27.5%, 51.9%; MCP-1 at 3h, 6h, 12h, 24h, respectively at each time point decreased 10.7%, 9.93%, 47%, 57.6%; there is no significant difference and statistically significant between given saline group and ox-LDL injury group. Western Blot Western blot analysis showed that: Compared with the control group in the ox-LDL stimulated HUVEC to inflammatory factors after ICAM-1, MCP-1 protein expression was significantly increased with ox-LDL compared injury Shuxuening injection given after ICAM-1, MCP-1 protein expression was significantly reduced. There was a significant difference (P <0.01); given saline had no significant difference compared ox-LDL injury, had not statistically significant. Conlusion(1) Ox-LDL has cytotoxic function on HUVEC and endothelial cells can cause structural damage;(2) Shuxuening injection can increase the oxidative damaged by ox-LDL's HUVEC proliferation activity (survival rate);(3) The role of ox-LDL up-regulated in HUVEC ICAM-1, MCP-1 protein expression. The Shuxuening injection can inhibit the ox-LDL-induced human umbilical vein endothelial cells (HUVEC) ICAM-1, MCP-1 protein upregulation. Then play on the protective effect of vascular endothelial cells, thereby preventing the occurrence of AS.