The Role Of Endothelial Microparticle And Its TF Pathway In Prevention Of Umbilical Vein Endothelial Cell Injury By Ginsenoside Rb1

Objectives: To investigate the effect of ginsenoside Rb1on monocrotaline-inducedapoptosis, formation of endothelial microparticles and expression of tissue factor inhuman umbilical vein endothelial cells, furthermore to explor the role of microparticleand its TF pathy way in ginsenoside Rb1-induced cytoprotective effects on MCTtoxicity in human umbilical vein endothelial cells.Methods:1. Human umbilical vein endothelial cells (EA.hy926cells) were culturedin vitro;2. In Control group, no drugs were treated with cells; In Rb1160group: cellswere treated with160mg/L ginsenoside Rb1only; In MCT group: MCT was addedto the culture medium to a final concentration of7.5mM; In MR40, MR80,MR160andMR200groups, Cells were pretreated with40,80,160or200mg/L ginsenoside Rb1for2h respectively and then exposed to7.5mM MCT for48h.3. Apoptosis ofEA.hy926cells was determined by fluorescence microscope and flow cytometryanalysis, EMPs were evaluated by laser confocal fluorescence microscopy and flowcytometry analysis respectively, the expression of TF in culture medium was detectedby ELISA analysis, then the expression of TF in cells and EMPs was evaluated byWestern Blot analysis.Results: A certain concentration of MCT induced endothelial cells injury.7.5mM MCT induced apoptosis, promoted formation of endothelial microparticles.Increased expression of TF in the culture medium and significantly promotedexpression of TF in cells and EMPs. However, pretreatment of40,80,160or200mg/L ginsenoside Rb1for2h significantly reduced their levels, ginsenoside Rb1dose-dependently inhibited effect of MCT-induced cells injury. Pretreatment of40mg/L ginsenoside Rb1inhibited apoptosis, and had no significant difference comparedwith control; The number of EMPs treated with80mg/L ginsenoside Rb1had nosignificant difference compared with control group; Application of160mg/Lsignificantly decreased expression of TF, but had significant difference comparedwith control group still;160mg/L ginsenoside Rb1reduced expression of TF in cellsand EMPs, and had significant difference compared with MCT group; In200mg/Lginsenoside Rb1pretreatment group, the number of EMPs was lower than160mg/Lginsenoside Rb1pretreatment group, but had no significant difference.Conclusions: A certain concentration of MCT could induce apoptosis, promotesexpression of endothelial injury makers such as EMPs and TF. A certain concentrationof ginsenoside Rb1could inhibit apoptosis, inhibit formation of endothelialmicroparticles totally, decrease the expression of TF in cells and EMPs significantlyand reduce the level of TF to some extent. Ginsenoside Rb1shows protective effecton MCT-induced endothelial injury via endothelial microparticle and its TF pathway.