The Effects Of Antisense Oligodeoxynucleotide Of ICAM-1 And E-selectin On The Adhesion Of Human Umbilical Vein Endothelial Cells To Peripheral Blood Mononuclear Cells

Objective:To investigate the effects of antisense oligodeoxynucleotide (asODN) of ICAM-1 and E-selectin and the cooperative effects of them combining simvastatin on the expression of ICAM-1 and E-selectin in human umbilical vein endothelial cells(HUVECs) and their adhesion to peripheral blood mononuclear cells(PBMC).Methods:Primary HUVECs were cultured from human umbilical vein, the asODNs of ICAM-1 and E-selectin were synthesized and transducted into HUVECs by using liposome transfection technique. The experiment were divided into eight groups.there are basal group,TNF-αtreated group,liposome group,liposome-control asODN group,nude asODN group,lipo-asODN group,simvastatin group and the combining application of lipo-asODN and simvastatin group. HUVECs in all the seven groups execept basal were stimulation with TNF-α,then the expression of ICAM-1 and E-selectin in the HUVECs were measured by flow cytometry and real-time polymerase chain reaction respectively. The adhesion rates of HUVECs to PBMC were measured by cell adhesion experiments. Based on the above experiments , the endothelial cells were first treated with simvastatin and then transfected by asODN of ICAM-1 or E-selectin. The expression of ICAM-1 and E-selectin in the HUVECs and the adhesion rates of the HUVECs to PBMC were observed by a series of measurements.Results:⑴Compared to the TNF-αtreated group , the protein and mRNA expression of E-selectin and ICAM-1 in the HUVECs were remarkablely decreased in both E-selectin/ ICAM-1 asODN group and E-selectin/ ICAM-1 lipo- asODN group;⑵either the combining application of E-selectin lipo- asODN and simvastatin or ICAM-1 lipo- asODN and simvastatin can lower the protein and mRNA expression of E-selectin and ICAM-1 also;⑶the expression of E-selectin in the lipo- E-selectin asODN/simvastatin treated group was significantly decreased when compare to lipo- asODN group (p<0.01), but the expression of ICAM-1 was not significant change, it demonstrate that the combining application of E-selectin lipo- asODN and simvastatin have more effects.⑷Compared to the TNF-αtreated group, the adherent rate of HUVECs to PBMC was markedly decreased in lipo- asODN group and lipo- asODN / simvastatin group (p<0.01), especially the adherent rate of HUVECs to CD3+ cells. But no significant change can be observed between lipo- asODN group and lipo- asODN / simvastatin group(p>0.05).Conclusion:Both E-selectin asODN and ICAM-1 asODN can inhibit the protein and mRNA expression of the relative E-selectin and ICAM-1 in HUVECs, and decrease the adherent rate of HUVECs to PBMC either, especially the adherent rate to CD3+ cell. When combining simvastatin,the effect of lipo- asODN can be enhanced.This study provides evidence for the application of ICAM-1 asODN and E-selectin asODN in clinic for antiartherosclerosis gene therapy.