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 five groups based on the concentration of PMPs. The concentration were: 0, 10 μg/ml, 30 μg/ml, 50 μg/ml, 100 μg/ml, each group contained four wells. Cells in wells were collected after 4 h. The second part was didied into three groups based on the time of co-cultivation: 2 h, 4 h, 24 h, and the concentration of PMPs that were added into each wells was 50 μg/ml. Each group contained four wells. Cells in wells were collected after 2 h, 4 h, 24 h respectively. 3. The RNA of cells was extracted. Semiquantitate reverse transcription-PCR (SQRT-PCR) was used to detect the relative expression of E-selectin, I CAM-1 and VCAM-1 respectively.
Results In this study, we found that cultured HUVEC (CRL-1730) expressed E-selectin, ICAM-1 and VCAM-1 mRNA in basic states. The expressed levels of E-selectin, ICAM-1 and 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-1 and 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-selectin, ICAM-1 and VCAM-1 on HUVEC (CRL-1730). It may explain a possible mechanism of PMPs in coronary heart disease.
- Platelet microparticles
- human umbilical vein endothelial cell
- cell adhesion molecule
- ischaemia –reperfusion injury
Statistics from Altmetric.com
If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.