The Role Of Fractalkine In ICAM-1Expression Of Human Umbilical Vein Endothelial Cell Induced By Angiotensin Ⅱ

Objective:To explore the expression of fractalkine and ICAM-1proteins of human umbilical vein endothelial cells (HUVECs) induced by angiotensin Ⅱ in different concentrations and durations. Using small interfering RNA to silent FKN gene of HUVECs, and to investigate the mechanism of FKN protein in ICAM-1expression of human umbilical vein endothelial cell that induced by angiotensin Ⅱ.Methods:Cultured HUVECs were incubated with AngⅡ in two concentrations (10-7mol/l,10-6mol/l) for Ohour,12hours,24hours and48hours. The change of cell viability was detected through methyl thiazole tetrazolium (MTT), ICAM-1and FKN protein expression were measured by Western blotting. According to the FKN mRNA sequence, to establish three strings of FKN siRNA; FKN mRNA expression of HUVECs were detected by Real Time quantitative PCR after transfected with siRNA of FKN by lipofectamine2000for24hours; Using the most effective string of FKN siRNA, and then transfecting experimental HUVECs for6hours; Transfected Cells were incubated with AngⅡ(10-6mol/l) for24hours, FKN mRNA expression were detected by Real Time quantitative PCR, ICAM-1and FKN protein expression were measured by Western blotting.Results:1) Cell viability was decreased after AngⅡ incubation; AngⅡ of10-6mol/l was more effective in attenuating cell viability during the same intervention time; Cell viability decreased in a time dependent manner during the same concertration of AngⅡ intervention;2) FKN and ICAM-1protein expression were enhanced after AngⅡ incubation, AngⅡ of10-6mol/l was more effective in increasing FKN and ICAM-1protein expression, Both FKN and ICAM-1expression increased in a time dependent manner during AngⅡ incubation;3) FKN siRNA transfection can inhibit cells FKMmRNA expression in a great extent. Though transfected HUVECs was incubated with AngⅡ (10-6mol/l) for24hours, FKN and ICAM-1proteins expression were largely decreased.Conclusion:1) AngⅡ can affect HUVECs viability and enhance FKN and ICAM-1protein expression;2) Silence FKN gene can greatly attenuate FKN mRNA expression;3) FKN protein may participate in the AngⅡ-ICAM-lpath way.