Effect Of Platelet Micropartiles On The Expression Of Cell Adhesion Molecule In Endothelial Cell

We pay more attention to the clinical significance of platelet microparticles(PMPs) because the inspecting technology had been improved for the passed few years.The effect of PMPs in myocardial ischemia and ischemia-reperfusion injury (I/R injury) also became hotspots.The majority of studies indicated that the level of PMPs increased in I/R injury,but the mechanism was not completely clear yet.The studies about the relationshipes between PMPs and cell adhesion molecules were few,and the results of the studies were inconsistent.In the study, the cultured Human Umbilical Vein Endothelial Cells(HUVEC) CRL-1730 were interfered by PMPs of different concentration. The expression levels of E-selectin,ICAM-1 and VCAm-1 mRNA were observed by RT-PCR.The study may show the effect of PMPs on CAMs of HUVECs(CRL-1730).It may lay a founation to further study the role of PMPs on coronary heart disease.Objective:To examine the expressions of cell adhesion molecule(E-selectin.VCAM-1.ICAM-1) in HUVECs(CRL-1730),which is affected by platelet microparticles(PMPs).To investigate the effects of platelet microparticles in coronary heart disease.Methods:1. PMPs was extracted from anticoagulated blood with sodium citrate.The purity of PMPs was measured by flow cytometry.2. The prepared PMPs and CRL-1730 cell were co-cultured.The first part was divided into 5 groups based on the concentration of PMPs:.The concentration were:0,10μg/ml,30μg/ml,50μg/ml,100μg/ml,Each group contained 4 wells.Cells in wells were collected after 4h.The second part was didied into 3 groups based on the time of co-cultivation:2h.4h.24h,and the concentration of PMPs that were added into each wells was 50μg/ml.Each group contained 4 wells. Cells in wells were collected after 2h.4h.24h respectively.3. The RNA of cells was extracted. Semiquantitate reverse transcription-polymerase chain reaction(SQRT-PCR) was used to detect the relative expression of E-selectin,I CAM-1 andVCAM-1 respectively.Results:In this study,we found that cultured HUVEC(CRL-1730) expressed E-selectin,ICAM-1和VCAM-1 mRNA in basic states.The expressed levels of E-selectin,ICAM-1和VCAM-1 were increased when HUVEC(CRL-1730) were interfered by PMPs of centain concentration (p<0.05).But the PMPs stimulated HUVECs(CRL-1730) at different times,the expressions of E-selectin,ICAM-1and VCAM-1 were of no diffeence (p>0.05).Conclusions:1.The high purity of PMPs were successfully prepared in the study.2.The PMPs may increase the expressions of E-selectn,ICAM-land VCAM-1 on HUVEC(CRL-1730).It may explain a possible mechanism of PMPs in coronary heart disease.